International Journal of Plant Biology doi: 10.3390/ijpb17090096

Authors: Yu Lyu Wenbing Han

Plant homeobox domain (PHD) proteins are a class of zinc-finger proteins that play pivotal regulatory roles in plant responses to environmental stresses. Research on the identification and analysis of PHDs in Gossypium tomentosum (G. tomentosum) remains limited. The present study conducted a genome-wide analysis of the PHD members in G. tomentosum and examined their properties and functions. A total of 71 PHDs have been identified, designated as GtPHD1–GtPHD71, through a combination of profile HMM-based searches, BLAST searches, and canonical features of PHD domains. Multiple sequence alignment of the G. tomentosum PHD domains revealed a characteristic signature: X(0,1)-C-X(1,2)-C-X(6,12)-[VLI]-X-C-X(0,2)-[DPK]-X(1,2)-C-X(4,5)-H-X2-C-X(9,18)-[YWF]-X-C-X2-C. A comprehensive investigation encompassing RT–qPCR, promoter analysis, and transcription factor analysis concluded that G. tomentosum PHDs likely play critical regulatory roles in response to abiotic stresses, including drought, hypoxia, and low temperature.

International Journal of Plant Biology doi: 10.3390/ijpb17090095

Authors: Mouna Ghorbel Kamel Jebreen Abdalmenem I. M. Hawamda Faiçal Brini

Non-specific lipid transfer proteins (nsLTPs) are small cysteine-rich proteins belonging to the cysteine-rich peptide (CRP) antimicrobial peptide superfamily, widely distributed across land plants and involved in key physiological processes including fertilization, cell wall organization, wax deposition, and responses to biotic and abiotic stresses. Recent genomic and evolutionary analyses have revealed an extensive diversification of nsLTPs across plant lineages, which has led to multiple classification systems based on molecular mass, cysteine spacing, intron structure, and GPI (glycosylphosphatidylinositol-anchored proteins)-anchor motifs. This diversity reflects their adaptive expansion during plant terrestrialization. This review proposes that nsLTPs act as central nodes connecting lipid metabolism, developmental signals, and stress response networks, functioning as integrative signaling hubs rather than simple lipid carriers. Building on this central thesis, we synthesize current knowledge on nsLTP structure, classification, and evolution, and examine their multifaceted roles in plant development and defense. We also highlight emerging biotechnological applications and identify key gaps that warrant further investigation.

International Journal of Plant Biology doi: 10.3390/ijpb17090094

Authors: Georgia Ouzounidou Niki-Sophia Antaraki Antonios Anagnostou George Daskas Ioannis-Dimosthenis S. Adamakis

Plant biostimulants may alter photosynthetic energy use and leaf structure, but their effects on complementary PSII energy partitioning in both cucumber leaves and fruits remain insufficiently characterized. Using plant material from a commercial greenhouse trial previously reported for agronomic outcomes, we evaluated a combined CropBioLife®, KeyPlex 120®, and Key Eco Oil® application protocol relative to conventional management. Chlorophyll Content Index (CCI), chlorophyll fluorescence imaging, and leaf anatomical traits were assessed. Under low light (LL; 455 μmol photons m−2 s−1), the combined-treatment greenhouse showed higher ΦPSII than the conventional-control greenhouse by 22% in fruits and 50% in leaves; fruit ΦNPQ was 17% higher, whereas ΦNO was 12% lower in fruits and 26% lower in leaves. Under high light (HL; 973 μmol photons m−2 s−1), differences in ΦPSII were smaller, while leaf ΦNO remained 17% lower. CCI was 38% higher in fruits and 52% higher in leaves. In leaves, spongy parenchyma thickness increased from 110 ± 3.4 to 135 ± 4.1 μm, palisade parenchyma thickness from 75 ± 2.1 to 78 ± 1.8 μm, total leaf thickness from 230 ± 4.6 to 245 ± 5.2 μm, and intercellular air-space area from 420 ± 18 to 610 ± 22 μm2. The parallel physiological and anatomical differences are consistent with a possible functional association, but gas exchange, mesophyll conductance, ROS, and water-use efficiency were not measured in the present dataset. Because the two management regimes occupied separate, non-replicated greenhouses and received different crop-protection inputs, the findings should be interpreted as treatment-associated differences rather than causal effects attributable to individual products.

International Journal of Plant Biology doi: 10.3390/ijpb17090093

Authors: Almagul Malimbayeva Azamat Khidirov Erbol Zhusupbekov Batyrgali Amangaliev Akerke Soltanayeva Aigul Raiymbekova Erbolat Berdibek Maksat Batyrbek

Sugar beet (Beta vulgaris L.) is an economically important industrial crop, and its productivity and sugar accumulation depend heavily on effective nutrient management. This study investigated how mineral fertilization combined with foliar applications of boron (B) and zinc (Zn) influences plant growth, photosynthetic performance, nutrient uptake, yield, and sugar production under irrigated alkaline soil conditions in southeastern Kazakhstan during the 2024 and 2025 growing seasons. The field experiment was conducted using a randomized complete block design with three replications and five treatments: (1) control (CK), (2) N300P150K300 (NPK), (3) NPK + B, (4) NPK + Zn, and (5) NPK + B + Zn. Mineral fertilization substantially enhanced plant growth and productivity compared with the unfertilized control, while foliar supplementation with B and Zn provided further improvements. Across both growing seasons, the combined NPK + B + Zn treatment consistently produced the greatest leaf area (55.6 × 103 m2 ha−1), dry biomass (4.9 t ha−1), photosynthetic potential (1631.5 × 103 m2 day ha−1), and net photosynthetic productivity (7.7 g m−2 day−1). It also resulted in the highest total uptake of nitrogen, phosphorus, and potassium, reaching 431.5, 64.7, and 761.4 kg ha−1, respectively. Foliar application of the combined micronutrients markedly increased their accumulation in plant tissues, with total B and Zn uptake rising to 874 and 1005 g ha−1, respectively. Sugar content improved from 17.1–17.2% in the control to 17.9–18.1% under the combined micronutrient treatment. As a result, the NPK + B + Zn treatment achieved the highest mean sugar yield (15.12 t ha−1), representing a 76.4% increase over the control and a 12.5% increase compared with NPK fertilization alone. The results demonstrate that foliar application of boron and zinc effectively complements mineral fertilization by enhancing photosynthetic activity, nutrient acquisition, and sugar accumulation. The combined application of NPK with foliar B and Zn can therefore be recommended as an effective nutrient management strategy for improving sugar beet productivity and sugar yield under alkaline soil conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17090092

Authors: Jenny B. Ryals Eric T. Stafne Haley N. Williams Patricia R. Knight

Improving stem cutting propagation of grapevine species (Vitis sp.) is important for supporting growing interest in southern grapevine production, particularly for lesser-known cultivars. More efficient and reliable propagation techniques can help nurseries increase the availability of grapevine plants for local growers and support development of the southern U.S. wine industry. Hardwood and semi-hardwood cuttings of four winegrape cultivars, ‘Mamont Noir’, ‘MidSouth’, ‘Miss Blanc’, and ‘Norton’, were subjected to 10 hormone treatments consisting of indole-3-butyric acid (IBA) alone or in combination with ascorbic acid (ASC). Treatments were applied as a three-second basal quick dip. Sixty days after treatment, total shoot and root number, average shoot and root length, rooting percentage, and cutting and root quality were evaluated. Based on the results, semi-hardwood or hardwood ‘Mamont Noir’ cuttings can be propagated using 50 ppm ASC or 50 ppm ASC + 500 ppm IBA. For nurseries where hardwood cuttings better fit the production schedule, ‘Miss Blanc’ cuttings rooted well regardless of hormone treatment. Additional research is needed to optimize hormone treatments for ‘MidSouth’ and ‘Norton’.

International Journal of Plant Biology doi: 10.3390/ijpb17090091

Authors: Khadijeh Shokri Naser Farrokhi Amir Mousavi Pär K. Ingvarsson Asadollah Ahmadikhah

Seed development in Brassica napus is tightly regulated at many stages. Microproteins (miPs), small regulatory peptides derived from larger ancestral proteins, have emerged as key post-translational modulators in plants; however, their role in seed development remains poorly understood. Here, a genome-wide screen for putative miPs in B. napus was performed, identifying 32 candidates. Among these, MIF1 (BnaC06g35530D), with domain similarity to ZF-HD transcription factors (TFs), showed robust expression dynamics. The integration of RNA-seq profiling, qRT-PCR, co-expression network modeling, and protein–protein interaction revealed a strong regulatory association between MIF1 and its predicted TF partner, i.e., ZHD (BnaC01g16510D), particularly at the zygote and bending cotyledon stages. Promoter analysis identified shared cis-elements among co-expressed genes, while structural modeling supported a stable MIF1–ZHD interaction interface. Evolutionary analysis demonstrated the conservation of zinc finger motifs, syntenic neighborhoods, and purifying selection on MIF1 across Brassicaceae species. Collectively, our findings suggest that MIF1 potentially functions as a conserved post-translational repressor of ZHD during seed development, contributing to fine-tuned transcriptional regulation in polyploid oilseed crops. This study provides a testable hypothesis about the interaction between MIF1 and ZHD and also paves the way for the future functional characterization of miP-TF modules in plant development.

International Journal of Plant Biology doi: 10.3390/ijpb17090090

Authors: Jolyon Dodgson Nicole Pereira John Clarkson Anna K. Marks David J. Marks

The marketable yield of carrot is reduced by carrot cavity spot disease (Pythium violae and P. sulcatum). In the UK, metalaxyl-M is the only approved fungicide. Good calcium nutrition can manage cavity spot, but calcium is not generally mobilised to the maturing carrot root. We report field-based studies testing the effect of Albina, a novel calcium functional fertiliser, which contains a calcium transport stimulant (MCAS) LoCalTM (Levity Crop Science, UK) that mimics the effect of auxin in relation to the transport of calcium across cell membranes. Its effect on cavity spot was compared with calcium nitrate (calcium fertiliser without MCAS) and metalaxyl-M in an inoculated ‘macrocosm’ plot experiment (cv. Nairobi), and in grower-managed field trials where it was compared with standard fertiliser and fungicide applications over two years (cvs. Nairobi and Octavo). In the macrocosm trial, the functional fertiliser gave significantly reduced cavity spot (12% incidence) compared with the inoculated (untreated) control (24% incidence) and had similar efficacy to the fungicide. In the grower-managed field trials, the functional fertiliser resulted in significantly improved cavity spot control with a mean reduction of 14%. The functional fertiliser could provide a cost-effective and environmentally friendly means for reducing cavity spot in carrots.

International Journal of Plant Biology doi: 10.3390/ijpb17090089

Authors: Maanda Ligavha-Mbelengwa Humbulani Munzhelele Peter Tshisikhawe Alphonce Bere John Ogola

Bush encroachment by woody species threatens biodiversity, ecosystem functioning, and rangeland productivity in savanna ecosystems. Understanding how juvenile woody plants respond to cutting at different times of the year is essential for developing effective management strategies. This study evaluated the seasonal resprouting responses of juvenile individuals of Dichrostachys cinerea and Terminalia sericea following cutting disturbances. The study was conducted in the Nylsvley Nature Reserve using line transects and plot-based sampling because of challenges associated with protecting experimental sites in communal lands. Juvenile plants of each species were allocated into groups comprising thirty-five individuals and subjected to seasonal cutting treatments. Resprout height, leaf production, and root sugar concentrations were measured to assess seasonal recovery patterns. Both species successfully resprouted after cutting regardless of season. T. sericea produced taller resprout heights than D. cinerea, whereas D. cinerea generally produced more leaves. Seasonal differences significantly affected recovery, with winter cutting producing the tallest resprout heights and greatest leaf production, while summer cutting resulted in the shortest resprouts and fewest leaves. Winter-cut plants produced more than twice as many leaves as summer-cut plants. Winter cutting drastically boosts plant regeneration capacity and cannot effectively curb bush expansion, whereas summer cutting achieves better suppression effects. Root sugar concentrations were lower during winter in both species, indicating reduced metabolic activity. Summer is the most effective season for cutting juvenile individuals of D. cinerea and T. sericea to suppress bush encroachment, whereas winter is the least effective. These findings provide evidence to support seasonally informed bush management strategies that promote biodiversity conservation, ecological integrity, and sustainable savanna ecosystem management.

International Journal of Plant Biology doi: 10.3390/ijpb17090088

Authors: Milton Sebastião Zavale Arsénio D. Ndeve Winfred N. Muteti Rogério M. Chiulele

Background/Objectives: Sweet orange (Citrus sinensis (L.) Osbeck) is an economically important fruit crop that contributes substantially to food security and smallholder income in Mozambique. Despite this, the genetic diversity of the locally grown germplasm has not been characterized at the molecular level, limiting its improvement and conservation programs. This study assessed the genetic diversity and population structure of germplasm from 94 sweet orange trees sampled across four districts of Inhambane Province using DArTSeq single-nucleotide polymorphism (SNP) markers. Methods: After filtering 8111 SNPs for call rate (≥0.80) and minor allele frequency (≥0.01), 1263 markers were retained, of which 1144 were anchored to the nine chromosomes of the reference genome. Results: Sparse non-negative matrix factorization identified K = 1, indicating a single undifferentiated gene pool, supported by a smooth PCA scree with one weak axis. DAPC assigned individuals to their district only 38.3% of the time (random expectation = 25%; maximum a-score = 0.10), and the first two PCoA axes explained 10.33% of variation with complete district overlap, indicating no detectable geographic structure. Diversity was low, with observed heterozygosity (Ho = 0.247) exceeding expected heterozygosity (He = 0.138) and a negative inbreeding coefficient (Fis = −0.222). The pattern indicated a heterozygote excess consistent with the fixation of the heterozygous interspecific-hybrid genome under clonal propagation. A hierarchical analysis of molecular variance showed that differentiation among districts was negligible (0.04%), whereas 4.16% of variation was partitioned among orchards (farms) within districts, indicating that the little of the existing structure resides at the orchard level, confounded with propagation method and cultivar, rather than among districts. Most variation was partitioned within individuals (76.0%), and pairwise FST values (0.0005–0.0035) were uniformly low. Conclusions: These results indicate that the sweet orange orchards stem from a single, highly heterozygous gene pool redistributed through the exchange of seed and vegetative planting material. This underscores the need to introduce diverse external germplasm to broaden the genetic base for sustainable improvement in Mozambique.

International Journal of Plant Biology doi: 10.3390/ijpb17090087

Authors: Maria Raquel Alcantara de Miranda Luiz Ferreira Coelho Junior Gabrielle Glayssa Lopes de Oliveira Holanda Rute Xavier Franca Elenilson Godoy Alves Filho Sueli Rodrigues Izabella Maria Costa Oliveira Fernando Antônio Souza Aragão Ricardo Antonio Ayub

This study investigated the differences between ripening patterns of two homozygous lineages of melons, yellow A5, a non-climacteric inodorus, and Charentais C1, a climacteric cantalupensis, regarding the physiological processes related to postharvest quality with a focus on carbohydrate metabolism and osmotic potential. Melons underwent significant changes as maturation progressed from 24 to 34 days after pollination (DAP) for the non-climacteric A5 lineage and from 18 to 28 DAP for the C1 climacteric lineage. The most discriminant variables for the non-climacteric pattern of A5 melons were the higher activity of cell wall hydrolase PME and lower firmness, the higher activity of enzymes involved in carbohydrate metabolism, and sucrose accumulation. In contrast, climacteric C1 melons reach physiological maturity at 24 DAP, do not accumulate sucrose, and enzyme β-GAL plays a key role in cell wall breakdown, contributing to softening and to soluble solids content. A5 melons exhibited greater TSS and soluble solids content, and even though the osmotic potential did not change significantly, the VIP results highlighted it as an important variable for discriminating between its maturation stages. Meanwhile, in the C1 melon, the increase in soluble solids content influences the more negative values of osmotic potential, not by changes in TSS content, and thus may be associated with fruit growth.

International Journal of Plant Biology doi: 10.3390/ijpb17090086

Authors: Karina A. Manakhova Lidiia S. Samarina Lyudmila S. Malyukova Lada V. Zhokhova Alexey V. Ryndin Evgeny I. Rogaev

Nitrogen (N) availability strongly influences tea growth and the accumulation of quality-related metabolites, but long-term responses to withdrawal of external N can vary among cultivars. In this study, we characterized phenotypic, spectral, biochemical, and transcriptomic responses of four Camellia sinensis accessions—‘Kolkhida’, ‘Karatum’, and γ-irradiation-derived mutant forms #582 and #619—during two, four, or six months of complete withdrawal of exogenous NH4NO3 in greenhouse sand culture. Phenotypic, spectral, and biochemical measurements included all four accessions at all three time points; RNA-seq data were available for all four accessions were represented at four and six months, whereas two-month point were available only for ‘Kolkhida’ and ‘Karatum’. Nitrogen withdrawal decreased L-theanine and caffeine and generally increased simple and gallated catechins, consistent with a change in carbon/nitrogen balance towards phenolic metabolism. The four cultivars exhibited markedly distinct responses. ‘Karatum’ showed the optimum resilience, with minimal phenotypic alteration and a neutral transcriptional response, while #582 demonstrated a stronger stress-associated phenotype and metabolic response. Within the available RNA-seq comparisons, DEG counts were lower at four months than at six months, while the two-month datasets for ‘Kolkhida’ and ‘Karatum’ contained the largest DEG sets for those two cultivars, suggesting a phased acclimation process. CsELIP1, CsSRG1, CsHHO2/4, CsNRT2.4, and CsTAT2 were identified as potential candidate genes associated with nitrogen limitation, flavonoid regulation, and amino acid metabolism. Our findings show that N-deficiency reactions in tea are highly cultivar- and time-dependent, making single-time-point evaluations insufficient. We propose ‘Karatum’ as a promising candidate for further evaluation in low-nitrogen dose–response and field trials based on our combined biochemical and transcriptional findings.

International Journal of Plant Biology doi: 10.3390/ijpb17090085

Authors: Aidana K. Argumbayeva Balnur R. Kali Laura S. Abeuova Beibit N. Nasiyev Aisulu Orken Dilnur Tussipkan Shuga A. Manabayeva

Drought is a major limiting factor for potato productivity. This study examined two Kazakh potato cultivars, Tokhtar and Aksor, under osmotic stress induced by polyethylene glycol (PEG)-6000 using an integrated physiological, biochemical, and molecular approach. We evaluated plant growth, total chlorophyll content, catalase activity, malondialdehyde content, and the transcript levels of six stress-associated genes (StMYB306-like, StuNAC, TRE, OST2, and EPF1, and EPF2) across increasing PEG concentrations. PEG treatment significantly inhibited growth and reduced total chlorophyll content while increasing CAT activity and MDA accumulation. The magnitude and statistical significance of the morphological and biochemical responses varied between cultivars and among PEG concentrations. Transcript levels of StMYB306-like, StuNAC, TRE, and OST2 generally increased with increasing PEG concentration, while EPF1 and EPF2 levels decreased. The absence of significant cultivar × PEG interactions for all six genes indicated that their transcriptional responses to PEG did not differ statistically between Tokhtar and Aksor. Correlation and multivariate analysis revealed contrasting associations between growth and chlorophyll-related traits and CAT activity, MDA accumulation, and stress-responsive gene expression. To our knowledge, the expression patterns of TRE, OST2, and EPF1 and EPF2 in potatoes under PEG-induced osmotic stress have not been characterized previously. Taken together, these findings characterize the coordinated morphological, biochemical, and transcriptional responses of potato plantlets to PEG-induced osmotic stress. They also identify candidate response traits for validation across genetically diverse germplasm and under soil-based and field drought conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17090084

Authors: Miroslav Salaić Zvonimir Zdunić Antun Jambrović Domagoj Šimić Sonja Petrović Andrija Brkić Vlatko Galić

This study explored how nitrogen fertilization modulates the grain yield response of maize to atmospheric stress across distinct phenological windows. Field trials were conducted from 2023 to 2025 under three nitrogen regimes (N0, N69, and NFull) with 237 maize testcrosses. Atmospheric stress was quantified as the cumulative number of hours with a vapor pressure deficit (VPD) exceeding 3.0 kPa. Data were analyzed using linear mixed-effects models. The 15-day pre-silking window was identified as the most sensitive among the three evaluated phenological windows, exhibiting a strong negative scaled effect of −0.390. The interaction between nitrogen treatment and pre-silking stress was highly significant. Elevated VPD did not operate as an independent determinant of final yield, as the highest mean grain yield was achieved during the hydrothermally intense 2025 season (8.319 t ha−1). NFull effectively mitigated the physiological impact of stress, securing the shallowest yield decline slope (−0.038 t ha−1 per hour). Conversely, the intermediate N69 treatment exhibited the highest statistical sensitivity (−0.109 t ha−1 per hour), reflecting phenotypic plasticity. Random-effects analysis revealed substantial genetic variability for pre-silking VPD sensitivity. The lack of correlation between absolute yield potential and stress sensitivity enables simultaneous selection for both high yield and phenotypic stability within this germplasm.

International Journal of Plant Biology doi: 10.3390/ijpb17090083

Authors: Sergio Rubén Pérez-Ríos Nayla Tamara Sánchez-Granados Juan Ocampo-López Mariana Saucedo-García Fabián Fernández-Luqueño María Fernanda Cenobio-Galindo Macario Vicente-Flores Ma Isabel Reyes-Santamaría Cristián Raziel Delgado-González Iridiam Hernández-Soto

Lilium sp. is one of the most commercially important ornamental species; however, improving its ornamental quality remains a challenge for production systems. In this context, nanoparticles have been proposed as an innovative alternative to optimize the agronomic performance of ornamental crops. The objective of this study was to determine how the foliar application of selenium nanoparticles (SeNPs) and titanium dioxide nanoparticles (TiO2NPs) modifies the physiological response and stem morphoanatomy of Lilium cv. Concador. Plants were grown under greenhouse conditions using conventional agronomic management for commercial Lilium production. We evaluated vegetative growth variables, relative chlorophyll content (SPAD), root biomass, flowering characteristics, and quantitative stem anatomy using histological, morphometric, and multivariate statistical analyses. Most agronomic variables, including plant height, number of leaves, shoot biomass, and flower opening, did not differ significantly between treatments, indicating that the nanoparticles did not compromise vegetative development. In contrast, root biomass and stem anatomical organization responded differently depending on nanoparticle type and applied dose. SeNPs, particularly at the highest dose, promoted greater development of parenchyma and vascular tissue, while TiO2NPs induced dose-dependent anatomical responses, with less parenchyma expansion and less pronounced vascular remodeling. Taken together, these results demonstrate that foliar application of nanoparticles induces early remodeling of the stem’s functional architecture without affecting vegetative growth and show that quantitative anatomy is a sensitive tool for detecting early responses to nano-inputs in ornamental species.

International Journal of Plant Biology doi: 10.3390/ijpb17090082

Authors: Juan Camilo Alvarez-Diaz Daniela Marin Mariana Espinal-Guzman Esteban Felipe Loaiza-Jaramillo Mario Alberto Quijano-Abril

Biological invasions threaten tropical biodiversity, yet the molecular mechanisms underlying successful invaders remain poorly understood. Thunbergia alata (Black-eyed Susan) is an aggressive “genomic orphan” invader in Andean forests, causing significant ecological disruption. This study contributes as one of the few transcriptomic profiles of light vs. shade responses in T. alata, providing a foundational molecular resource for this species. We performed differential expression analysis under varying light conditions, revealing a massive transcriptomic shift involving over 4000 differentially expressed genes. Our findings suggest a robust and stable homeostatic adaptation mechanism, characterized by the up-regulation of SnRK1 subunits for energy sensing and strategic management of Reactive Oxygen Species (ROS) within the thylakoid membrane. Furthermore, the fine-tuning of PIF/Auxin modules and the condition-specific induction of NAC and WRKY transcription factors facilitate shade avoidance and rapid vertical growth. The identification of a substantial reservoir of species-specific “not classified” genes suggests that novel genetic elements contribute to T. alata’s adaptive success. By elucidating these key regulatory networks, this research provides an important genomic baseline for future studies on adaptive evolution and the development of molecularly informed strategies for managing and controlling this invasive species in new environments.

International Journal of Plant Biology doi: 10.3390/ijpb17090081

Authors: Ha Duc Chu Trung Quoc Nguyen Anh Quynh Ho Gioi Huy Dong Hong Viet La Nguyen Thi Phuong Thao Linh Hong Ta

Single-nucleotide polymorphism (SNP) arrays provide reproducible, accurate, and scalable genotyping for rice research and cultivar development. This review critically evaluates the design and development of major rice SNP platforms and defines their value within a rapidly changing genotyping landscape. In contrast to earlier platform-centered summaries, this review connects array selection with the complete breeding pathway from germplasm assessment and locus discovery to functional validation, marker-assisted transfer, genomic selection, varietal authentication, and pre-breeding. SNP arrays are compared with genotyping-by-sequencing, double-digest restriction site-associated DNA sequencing, low-pass sequencing, whole-genome resequencing, targeted sequencing, and long-read sequencing in terms of cost, throughput, data completeness, computational demand, marker discovery, and population transferability. Candidate-locus validation through gene-expression analysis, haplotype analysis, near-isogenic lines, RNA interference, transgenic complementation, clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) editing, and field evaluation is also examined. Pangenome resources and the contributions of structural variants, presence/absence variations, and copy number variations to probe design and trait analysis receive particular attention. The review further assesses ascertainment bias, weak detection of rare alleles, genotype-cluster errors, limited portability across distant germplasm, and reduced genomic prediction across populations or environments. Future priorities include multi-omics integration, climate-resilient rice improvement, modular panels, imputation anchor loci, and economical deployment through low-density assays, shared facilities, and kompetitive allele-specific polymerase chain reaction conversion. The value of SNP arrays is greatest when stable genotype calls, large sample numbers, and repeated use across breeding cycles are required. Their practical contribution should be judged by prediction accuracy, selection cost, cycle duration, donor-segment control, realized genetic gain, and cultivar deployment.

International Journal of Plant Biology doi: 10.3390/ijpb17090080

Authors: Lyna Miloudi Khaled Rezeg Mohamed Kotoub Miloudi

Distinguishing crop from weed species at the seedling stage is a fine-grained morphological discrimination problem. We study it on the public Plant Seedlings benchmark, which contains twelve species (three crops and nine weeds) imaged at the seedling stage. Automated classifiers report near-ceiling accuracy on this benchmark. Yet its images are acquired in controlled trays containing soil, gravel, rulers, barcodes, and printed labels, so a model may identify a species from its acquisition context rather than its morphology. We audit two architectures, EfficientNet-B7 and ViT-B/16, trained on the V2 dataset (5539 images) and probed with plant-only, background-only, and background-swapped inputs. Near-ceiling models (96.1% and 96.4% over three seeds) recover the correct species for up to 47.7% of samples from the background alone (chance 8.3%) and lose about 60 points under background swapping. Context reliance is therefore a property of the benchmark, not any single architecture. Tracing this to its source, an independent learned representation of the plant-free background alone identifies the species at 72.4%. The reliance is correctable end to end: a consistency-regularisation scheme retains 92.1% full-image accuracy for the transformer with no segmentation at inference, at an architecture-dependent cost. Reported accuracy thus partly measures acquisition context, not morphology; morphological grounding should be measured and reported alongside accuracy.

International Journal of Plant Biology doi: 10.3390/ijpb17090079

Authors: Efthymios Poulios Sousana K. Papadopoulou Evmorfia Psara Agathi Pritsa Constantinos Giaginis

Background/Objectives: Medicinal and aromatic plants are important sources of phenolic compounds with antioxidant properties. This study evaluated the total phenolic content (TPC) and antioxidant activity of 19 commercially available medicinal herb and spice products of reported Mount Athos provenance and investigated the influence of extraction solvent on these properties. Methods: Extracts were prepared by ultrasound-assisted extraction using water, 50% aqueous methanol, or 50% aqueous acetone. TPC was determined using the Folin–Ciocalteu assay, while antioxidant activity was evaluated using DPPH, ABTS, FRAP, and a qualitative supercoiled plasmid DNA relaxation assay. Solvent effects were assessed using Friedman and post hoc Wilcoxon signed-rank tests, and associations between TPC and antioxidant activity were examined using correlation and regression analyses. Results: TPC and antioxidant activity varied considerably among botanical products and extraction conditions, with Origanum spp. generally exhibiting among the highest TPC values and strong antioxidant responses. Aqueous acetone yielded significantly higher TPC than aqueous methanol and water. Both aqueous organic solvents produced greater DPPH inhibition than water, whereas aqueous methanol produced greater ABTS activity than water; no significant solvent effect was observed for FRAP. TPC was positively associated with antioxidant activity, particularly ABTS and FRAP, explaining up to 78% of its variability. Several extracts also preserved the supercoiled plasmid DNA form under hydroxyl radical-generating conditions, although this observation requires quantitative confirmation. Conclusions: Medicinal herbs and spices of Athonite provenance exhibit considerable phytochemical and antioxidant variability. Extraction solvent influences phenolic recovery and antioxidant activity in an assay-dependent manner, with no single solvent being uniformly superior. These findings provide baseline data supporting further phytochemical characterization and quantitative evaluation of the antioxidant and DNA-protective properties of these botanical materials.

International Journal of Plant Biology doi: 10.3390/ijpb17090078

Authors: Janyne Soares Braga Pires Francine Bonomo Crispim Silva Maria Eduarda da Silva Barbosa Geovana Ribeiro Cavilha Mateus Moura Coelho Samile Mardegan Otilia Fernando Gomes Hoste Ana Júlia Câmara Jeveaux-Machado Lúcio de Oliveira Arantes Vinicius de Souza Oliveira Simone Alves Fernandes Johnatan Jair de Paula Marchiori Adriano Alves Fernandes Sara Dousseau-Arantes

Water deficit is one of the main limiting factors for crop establishment and productivity, particularly during seed germination and early seedling development. This study aimed to evaluate the biostimulant potential of Ascophyllum nodosum extract applied to maize (Zea mays L.) seeds subjected to osmotic stress induced by polyethylene glycol (PEG-6000). Three independent bioassays were conducted under controlled conditions. In the first assay, osmotic potentials ranging from 0 to −0.8 MPa were evaluated to characterize stress levels and identify conditions representative of moderate and severe water deficit. In the second assay, increasing doses (0 to 2.0 mL kg−1 of seeds) of a commercial A. nodosum-based extract and its isolated mineral fraction were evaluated to determine the optimal application range. In the third assay, the selected treatment was evaluated under control conditions and under moderate (−0.6 MPa) and severe (−0.8 MPa) osmotic stress. Germination percentage, normal and abnormal seedlings, radicle and epicotyl length, and seedling vigor index were assessed. Osmotic stress progressively reduced germination and seedling growth, with the strongest effects observed at −0.6 and −0.8 MPa. Seed treatment with A. nodosum extract did not impair final germination but improved seedling development in a dose-dependent manner. The highest efficiency was observed at intermediate doses, with maximum normal seedling percentage predicted at 0.45 mL kg−1 and maximum radicle growth at approximately 0.66 mL kg−1. Under water deficit conditions, the complete extract promoted greater improvements in seedling performance than the mineral fraction, indicating that the beneficial effects were mainly associated with bioactive organic compounds rather than mineral nutrition alone. These findings provide new evidence that dose optimization is essential for maximizing the efficiency of seaweed-based biostimulants and demonstrate the potential of A. nodosum seed treatment as a sustainable strategy to improve maize establishment under water-limited conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17080077

Authors: Jovana Eskildsen Tobias Hanak Rebecca Hood-Nowotny Magdalena Musialak-Lange Ewelina Sokolowska Sylwia Kierszniowska Claus Krogh Madsen Inger Holme Henrik Brinch-Pedersen

Dwarf cereal cultivars were crucial for the Green Revolution. Dwarfed, lodging-resistant varieties remain essential today, as climate change brings more storms and downpours. In barley, the dwarfing gene HvDEP1 has been widely used in breeding. Although its pleiotropic effects on agronomic traits have been examined, previous studies relied on cultivars developed via random mutagenesis, which carry background mutations that may influence phenotypes. Moreover, its impact on root traits remains underexplored. We used CRISPR/Cas9 to generate precise HvDEP1 mutants and introduce dwarfism into the barley cultivar ‘Maythorpe.’ We assessed the effects on above-ground morphology, yield-related traits, root architecture, biomass via 13C labelling, and the root metabolome. HvDEP1 mutations significantly reduced plant height, straw, spike, and awn length, as well as thousand-grain weight. An in-frame mutant showed intermediate height, straw, and awn phenotypes. Belowground, in a root experiment restricted to knockout line #12, specific root length and the length of the finest (0–0.25 mm) roots were reduced, while total root length was lower but not significantly so; (p = 0.062). Root metabolomic profiling detected no genotype-associated differences. These results provide new insights into HvDEP1′s role in both shoot and root systems and demonstrate that precise CRISPR/Cas9-mediated editing can rapidly introduce dwarfism while revealing trade-offs in other agronomic traits.

International Journal of Plant Biology doi: 10.3390/ijpb17080076

Authors: César Augusto Ticona-Benavente Erica Ines Almeida de Souza Álvaro Brasil Barbosa Neto Jean Piere Jesús Chicana Marina José Ramirez-Chung Johnny Campos-Cedano Jorge Villacres-Vallejo

In Amazonian Spodosols, soil acidity and nutrient constraints limit the cultivation of cutleaf groundcherry. One sustainable alternative for cultivating this species is the use of Trichoderma inoculants, but studies on their use in this crop are scarce. This study evaluated whether rice-based inocula of Trichoderma harzianum (TH) CCB-LA101 and T. viride (TV) CCB-LA103 improved growth and fruit production of cutleaf groundcherry and whether the responses varied with the application dose. Plants were grown in a randomized complete block design with a 2 × 3 factorial arrangement: two Trichoderma species and three inoculum doses applied to the soil (0, 4, and 8 g plant−1; Trichoderma concentration of 3.2 × 109 CFU g−1). Species and dose affected fruit width, length, number, and yield, with TH more effective than TV at increasing fruit number and yield. The contrast between inoculated and non-inoculated plants was significant for all traits. The 8 g plant−1 dose significantly increased fruit number and yield by 22.8% and 17.7%, respectively, relative to the non-inoculated control, whereas fruit width and fruit length decreased by 8.3% and 7.3%, respectively. Considering the six treatment combinations, the 8 g plant−1 TH inoculum (TH8) was the only treatment to significantly increase fruit yield, with net returns for smallholders estimated at USD 1104.46 ha−1. Therefore, under the tested conditions, TH8 could be used to grow cutleaf groundcherry in Amazonian Spodosols.

International Journal of Plant Biology doi: 10.3390/ijpb17080075

Authors: Antonio Samudio Oggero Daisy Ramírez Monzón Héctor D. Nakayama Luis Felipe Medeiro Alves Valter Arthur Oscar Vega Alvarenga Gloria A. Resquín Romero Wilson Romero Vergara Juan D. Avalos Añazco

Calibrating the mutagenic dose is the first practical step of any radiation mutation-breeding programme, and it is usually summarised by the median lethal dose (LD50) or the median growth-reduction dose (GR50). We asked whether an accessible, image-based phenotyping pipeline can quantify the early radiation response of cowpea (Vigna unguiculata L. Walp.) seedlings finely enough to estimate GR50 and to rank organ- and pigment-level sensitivities. Seeds of the traditional Paraguayan landrace kumandá pyta’i were exposed to Cobalt-60 gamma rays at 0, 100, 200, 300, 400, 500, 600, and 700 Gy, grown in a greenhouse, and photographed at the early seedling stage. A single calibrated photograph (5.1 px mm−1) of 83 seedlings was segmented in Fiji/ImageJ 1.54p and analysed with Python to extract morphometric traits (total, root, and shoot length, root:shoot ratio, tortuosity, and a two-dimensional biomass proxy) and colorimetric traits (CIE L*a*b*, a normalised greenness index, and colour-class pixel fractions). Because the data departed from normality, dose effects were tested with Kruskal–Wallis, Spearman rank correlation, and Dunn post hoc tests, and GR50 was estimated by regression of each trait expressed as a percentage of the control. Total length, shoot length, and the biomass proxy declined significantly with dose (Spearman ρ = −0.40, −0.51, and −0.47; all p < 0.001), preceded by a low-dose stimulation at 100 Gy. Estimated GR50 values were ≈390 Gy for shoot length, ≈510 Gy for total length, and ≈550 Gy for the biomass proxy, within the range reported for other cowpea genotypes. Shoot elongation was more radiosensitive than root elongation, so the root:shoot ratio did not decline; tortuosity showed no dose response. Among pigment traits, the loss of greenness was the most robust signal (a* increased, ρ = +0.62, p = 5 × 10−10; green pixel fraction fell from 0.32 to near zero by 500 Gy). These results show that single-photograph phenotyping resolves a coherent, statistically supported dose response and yields a GR50 estimate usable for dose calibration. For kumandá pyta’i, doses of roughly 300–400 Gy (below GR50) are the most defensible starting window for mutation induction. The framework is reproducible and low-cost, but it is based on one greenhouse experiment and a single genotype, and should be validated across independent trials and cultivars.

International Journal of Plant Biology doi: 10.3390/ijpb17080074

Authors: Awais Ali Md Noor E Azam Khan Nishma Dhakal Fahmida Fiza Zhiheng Xing Joseph Masabni Genhua Niu

Climate change is increasing the frequency and severity of abiotic stresses, including salinity, drought, heat, and heavy metal toxicity, which strongly threaten productivity, quality, and market value in horticultural crops. Although plant biostimulants have been widely reviewed in agronomic crops, their role in horticultural systems remains less comprehensively synthesized, particularly across different crop groups, stress types, application methods, and molecular response mechanisms. This review addresses this gap by systematically compiling current evidence on the use of biostimulants to improve abiotic stress resilience in horticultural crops, with particular emphasis on morphological, physiological, biochemical, and recently emerging molecular responses, especially transcriptomic evidence with supporting metabolomic information where available. Publications were retrieved from the Web of Science Core Collection using two searches covering 2016–2025 for morphological/physiological responses and 2021–2025 for molecular/genetic responses. Of 780 records initially identified, 134 studies met the inclusion criteria. Across these studies, the most frequently evaluated biostimulants were seaweed extracts, humic and fulvic substances, protein hydrolysates, and microbial inoculants, particularly PGPR and AMF. Biostimulant application consistently improved stress tolerance by enhancing antioxidant capacity, osmotic adjustment, nutrient use efficiency, cell wall strengthening, and hormonal regulation. Foliar applications were frequently used for rapid mitigation of drought- and heat-induced canopy-level physiological responses, whereas soil/root-zone application was more common for salinity and heavy metal stress. Emerging molecular evidence, dominated by transcriptomic studies and supported by limited metabolomic data, indicates that biostimulants may induce molecular priming through stress-responsive gene networks and associated metabolic adjustments. Overall, biostimulants show strong potential to improve abiotic stress resilience in horticultural crops, but broader adoption is constrained by variable efficacy, limited mechanistic validation, and inconsistent regulatory frameworks. Future research should prioritize multi-environment validation and functional genetics to support more reliable and targeted biostimulant use.

International Journal of Plant Biology doi: 10.3390/ijpb17080073

Authors: Amit Yadav Anuradha Singh Saurabh Pandey Jyotirmaya Mathan

Photosynthesis is the fundamental biological process underlying plant growth, crop productivity, and global food security. However, its efficiency is highly vulnerable to abiotic stresses, which disrupt chlorophyll biosynthesis, electron transport, carbon assimilation, stomatal regulation, and photoprotective mechanisms, ultimately reducing crop yield. Improving photosynthetic resilience under adverse environments has therefore become a major objective of modern crop improvement. Recent advances in phenomics and high-throughput phenotyping (HTP) have transformed the evaluation of photosynthesis-related traits by enabling rapid, non-destructive, and large-scale assessment across diverse environments, while facilitating quantitative characterization of structural, physiological, biochemical, and thermal responses to abiotic stress. Technologies including chlorophyll fluorescence, gas-exchange analysis, thermal imaging, hyperspectral imaging, LiDAR, and UAV-based sensing provide comprehensive insights into plant physiological responses and stress adaptation. Integration of these phenomic approaches with genomic information and artificial intelligence (AI)-driven analytical frameworks has strengthened genomic and phenomic prediction, enabling more accurate identification of candidate genes, selection of superior genotypes, and accelerated genetic gain. This review critically synthesizes recent advances in photosynthesis-related traits, phenomics, HTP technologies, and their integration with genomics and AI-assisted breeding, highlighting current challenges, knowledge gaps, and future opportunities for developing climate-resilient wheat and rice cultivars and promoting sustainable crop production.

International Journal of Plant Biology doi: 10.3390/ijpb17080072

Authors: Blanca Patricia Castellanos-Potenciano Erika Belem Castillo-Linares Eder Ramos-Hernández

The Annonaceae family in Mexico comprises a wide diversity of species with economic, cultural, agricultural, and pharmacological importance; however, research on this group remains fragmented. Therefore, this study analyzed the scientific output on Annonaceae in Mexico using a bibliometric approach. A total of 342 articles published between 1980 and 2025 in the Web of Science were retrieved and analyzed using VOSviewer version 1.6.16. The results showed a sustained increase in annual scientific production; approximately 90% of the articles were published in Q1 journals, with impact factors ranging from 1.3 to 13.4, highlighting the scientific relevance of this family. The main research areas were plant sciences, agriculture, biochemistry and molecular biology, entomology, and food science and technology. Emerging topics focused on the antitumor potential of species such as Annona muricata, A. cherimola, A. coriacea, and A. purpurea. Research on Annonaceae in Mexico has expanded in a multidisciplinary manner, although limited interaction between different areas and institutions persists. This field offers opportunities for biodiversity conservation, sustainable use, and the development of biotechnological and pharmacological applications.

International Journal of Plant Biology doi: 10.3390/ijpb17080071

Authors: Nattawut Rodboot Sureerat Yenchon

Waterlilies (Nymphaea spp.) are ecologically and ornamentally important aquatic plants, yet their micropropagation remains challenging due to endophytic contamination and the recalcitrant nature of tuber explants. This study established an efficient in vitro protocol for Nymphaea pubescens using field-collected tubers. Surface sterilization with 0.2% mercuric chloride (HgCl2) for 15 min achieved complete sterility with 80% explant viability, and an “escape technique” of excising newly emerged shoots from the maternal tuber reduced latent bacterial contamination. Liquid Murashige and Skoog (MS) medium with 1.0 mg/L 6-benzylaminopurine (BAP), 0.1 mg/L thidiazuron (TDZ), and 0.5 mg/L 1-naphthaleneacetic acid (NAA) produced greater shoot multiplication (17.20 ± 2.53 shoots/explant) and elongation (1.03 ± 0.22 cm) than semi-solid medium (9.70 ± 2.11 shoots/explant; 0.70 ± 0.13 cm) (p < 0.01). Transfer to semi-solid maturation medium with glutamine and casein hydrolysate produced the highest survival rate (100%) and significantly improved shoot quality (p < 0.01), mitigating hyperhydricity from prolonged liquid culture. Rooting on semi-solid medium with 0.5 mg/L NAA yielded 90% rooting, and over 80% of acclimatized plantlets established successfully in the field within two weeks. These results provide a practical, reproducible protocol for conservation and mass propagation of N. pubescens and a foundation for future in vitro mutation breeding in ornamental waterlilies.

International Journal of Plant Biology doi: 10.3390/ijpb17080070

Authors: Prabhaharan Renganathan Kristina Borisovna Ukhatkina Makhmutov Almaz Ilya Isidorovich Van Erp Lira A. Gaysina

Commercial seaweed extracts (SWEs) are widely used as plant biostimulants; however, information on formulation-specific optimum dosages for greenhouse-grown Capsicum annuum seedlings is limited. This study evaluated the effects of five commercial SWE formulations applied at five dosages (1–5 mL L−1) on seedling growth, biomass accumulation, and root growth under greenhouse conditions. Morphological traits, biomass characteristics, and growth efficiency indices were assessed using univariate and multivariate statistical approaches to identify formulation-specific optimum application rates. Commercial SWE application significantly (p < 0.05) increased chlorophyll content, shoot and root growth, and fresh biomass, although the magnitude of the response varied among the formulations and application rates. The optimum dosages identified by the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) were ASCO at 2 mL L−1, AQUA at 3 mL L−1, BIO at 2 mL L−1, KAT at 3 mL L−1, and SAGA at 3 mL L−1. Principal component and Pearson correlation analyses further demonstrated coordinated improvements across multiple growth traits. These findings demonstrate that optimum seedling performance depends on formulation-specific rather than universal application rates. The identified optimum dosages provide practical guidance for selecting commercial seaweed biostimulant application rates in greenhouse C. annuum nurseries.

International Journal of Plant Biology doi: 10.3390/ijpb17080069

Authors: Said Bahoch Salahddine Chafiki Soumaya El Assri Abdessamad Elaasri Rachid Elaini Fouad Elame Ahmed Wifaya Mohamed Ouknin El Hassan Mayad Rachid Bouharroud Redouan Qessaoui

The two-spotted spider mite, Tetranychus urticae Koch, is a highly polyphagous and economically important pest affecting a wide range of crops worldwide. This study evaluated the potential of four rhizobacterial isolates—Leucobacter aridicollis (IQR1), Paenochrobactrum sp. (IQR2), uncultured bacterium IQR3, and marine bacterium AK6_052 (IQR5)—as candidate biological control agents against T. urticae. The rhizobacterial isolates were obtained from the tomato rhizosphere at the INRA experimental farm (Agadir, Morocco) and evaluated for their acaricidal and repellent activities against T. urticae. Corrected mortality and repellency index were assessed after 24, 48, and 72 h of exposure. All four isolates exhibited acaricidal activity, with IQR3 and IQR5 showing the greatest efficacy. Corrected mortality reached 69.9% with IQR3 at the highest concentration after 72 h of exposure. The isolates showed no statistically significant repellent activity. Under greenhouse conditions (108 CFU mL−1), IQR3 was associated with the lowest infestation incidence (64%), the lowest infestation severity (13.3%) and the lowest adult density (1.8 adults leaf−1); IQR5 showed a comparable trend. Overall, IQR3 and IQR5 represent promising rhizobacterial candidates for the sustainable management of T. urticae in tomato production. Further studies are needed to optimize their formulation, elucidate their mechanisms of action, and validate their efficacy under field conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17080068

Authors: Kornkanok Chamchusri Bhornchai Harakotr Panumart Rithichai Yaowapha Jirakiattikul

Knowledge of pollen viability, pollen longevity, and stigma receptivity in Zephyranthes remains limited despite their importance for successful hybridization. Therefore, this study evaluated these reproductive traits in four Zephyranthes species (Z. traubii, Z. candida, Z. grandiflora, and Z. citrina) and one cultivar, ‘Pim Chomphu’, to provide baseline reproductive information for controlled pollination. Pollen viability was evaluated using tetrazolium staining at different collection times and storage conditions, whereas stigma receptivity was assessed using 6% hydrogen peroxide (H2O2), with reproductive outcomes following controlled self-pollination evaluated at the corresponding time points. Tetrazolium-based pollen viability exhibited taxon-specific temporal patterns; however, viability was generally high at 11:00, corresponding to 1 day after anthesis (1 DAA) in Z. traubii and the day of anthesis (0 DAA) in the remaining taxa. The combined protocol of desiccation followed by storage at −5 °C or −20 °C maintained tetrazolium-based pollen viability for up to 120 days. Although all taxa showed positive H2O2-based stigma responses across the evaluated time points, reproductive outcomes following self-pollination differed markedly among taxa. Z. citrina set fruit at all pollination times, whereas Z. candida failed to set fruit. The remaining taxa showed variable fruit set and seed germination depending on pollination timing. These results indicate that positive H2O2-based stigma responses should be interpreted as biochemical indicators of potential stigma receptivity and considered together with subsequent reproductive outcomes. These findings provide baseline reproductive information to support pollen management and controlled pollination in Zephyranthes.

International Journal of Plant Biology doi: 10.3390/ijpb17080067

Authors: Aidyn Orazov Talant Samarkhanov Anar Myrzagaliyeva Moldir Yermagambetova Sultan Kauanov Yerlan Turuspekov Serik Irsaliyev Shynar Tustubayeva Bauyrzhan Turalin

Wild almond relatives are valuable reservoirs of allelic variation for crop improvement and conservation, yet Kazakhstan’s wild-almond genetic resources remain poorly characterised. We conducted an exploratory SSR assessment of 80 putative individuals from four taxon-locality groups (20 per group), each representing one sampled population: Prunus ledebouriana, P. tenella, P. petunnikowii, and P. spinosissima. Of 22 nuclear simple sequence repeat loci screened for cross-taxon transferability, 15 generated reproducible profiles and were retained; their even genome-wide distribution was not verified. Across the full dataset, the mean number of alleles was 3.55, the effective number of alleles was 2.62, expected heterozygosity (He) was 0.544, and 95.0% of loci were polymorphic. Missing genotypes ranged from 0.0% to 34.7% among groups, and six loci had at least 20% missing data. AMOVA attributed 76.5% of variation to within-group differences and 23.5% to among-group differences (PhiPT = 0.235, p = 0.001). PCoA, unbiased Nei distances, UPGMA, and descriptive Bayesian clustering separated the four sampled groups. A nine-locus sensitivity analysis that excluded the six high-missing loci retained P. spinosissima as the group with the highest mean He (0.699), whereas P. petunnikowii increased from 0.471 to 0.609. Thus, the low full-panel estimate for P. petunnikowii was not robust to missing data. Because taxon identity was fully confounded with locality and the marker panel was limited, the results are interpreted as a regional marker-transferability and methodological baseline rather than as species-wide or genome-wide inference.

International Journal of Plant Biology doi: 10.3390/ijpb17080066

Authors: Mauricio Mateus Guimaraes-Silva Matheus Chaves de Jesus Dannily Augusto Rebouças Lorraynne Oliveira-Souza Abreu Claudete da Costa-Oliveira Elaine de Lima de Jesus Orlando Maia Barboza Tamara do Nascimento da Silva Philippe Barreto de Almeida Renato Crespo Pereira Davyson de Lima Moreira Ygor Jessé Ramos

Plant chemical diversity constitutes an essential dimension of biodiversity, arising from interactions among genetic background, biosynthetic regulation, the environment, ontogeny, ecology, and evolutionary history. Although the term chemotype is widely used for distinguishable chemical patterns within a taxon, its meaning remains inconsistent across plant biology, phytochemistry, pharmacognosy, chemical ecology, and natural-product standardization. This conceptual review examines the historical development, conceptual boundaries, chemical criteria, biological significance, and methodological requirements of plant chemotypes. We define a chemotype as a distinguishable, recurrent, relatively stable, analytically reproducible, and biologically interpretable intraspecific chemical pattern expressed by individuals, populations, or lineages of the same taxon. Assignment should be based primarily on chemical discontinuity, diagnostic markers, recurrence, relative stability, and analytical reproducibility; ecological, functional, biosynthetic, and genetic information should be treated as supporting evidence when available. The framework applies to volatile and non-volatile systems, including essential oils, cannabinoids, alkaloids, phenolics, and glucosinolates, while recognizing that platform coverage, annotation confidence, and taxon-specific biology can alter the inferred groups. We distinguish chemotypes from chemical phenotypes, chemical ecotypes, chemical geotypes, chronotypes, chemovars, and chemophenetic groups and propose three evidence levels: putative, supported, and confirmed chemotypes.

International Journal of Plant Biology doi: 10.3390/ijpb17080065

Authors: Aisulu Orken Nurbek Zhumabay Nazerke Amangeldyeva Malika Ramazanova Sabir Makhmadjanov Laura Tokhetova Shuga Manabayeva Dilnur Tussipkan

In arid and semi-arid regions, salinity and drought limit cotton productivity. As the northernmost cotton-growing country, Kazakhstan often deals with these issues. The objective of this study was to classify salt- and drought-tolerant groups within the Kazakhstan cotton collection based on key phenotypic traits. Fifty-six Gossypium hirsutum genotypes were evaluated under controlled conditions using NaCl and PEG-6000. Six morphophysiological traits, including germination rate, plant height, fresh weight, root dry weight, and relative water content, were analyzed. Increasing NaCl and PEG levels reduced vegetative growth, while germination remained relatively stable under moderate stress. Regression analysis identified 200 mM NaCl as the optimal salinity level for salt tolerance and 20–30% PEG as the optimal level for drought tolerance. Cluster analysis using membership function values grouped the genotypes into five tolerance categories. Eight lines including M-4016-6, M-4031-8, M-4014-6, M-4029-9, M-4016-7, M-4016-8, M-4020-2, and M-4016-4 exhibited high tolerance to both stresses. Additional lines exhibited combined tolerance, indicating strong breeding potential. Principal component analysis revealed an inverse relationship between salt and drought tolerance, suggesting stress-specific adaptation. This study is the first to report on the development of a classification system for salt and drought tolerance in the Kazakhstan Cotton Collection. The results demonstrate substantial genetic variability and highlight the potential for developing cotton cultivars adapted environments prone to stress.

International Journal of Plant Biology doi: 10.3390/ijpb17080064

Authors: Hiju Yoo Samuel Lee Young Jin Choi Yeong Sunwoo Jeong Geun Lee Siyeon Yoo Ji Min Seo Sunyoung Lee Gyurim Kim Eun Bin Cha Sang Yong Nam Jae Hwan Lee

Sorrel (Rumex acetosa L.) is a promising leafy vegetable for controlled-environment production, but integrated information on its growth, structural quality, optical traits, and photochemical responses to light-emitting diode (LED) spectra remains limited. To our knowledge, this study provides the first systematic comparison of five contrasting LED spectral conditions in sorrel using morphological, biomass, structural-quality, reflectance-based, and chlorophyll-fluorescence indicators. Plants were grown for six weeks under red light (RL), green light (GL), blue light (BL), purple-phyto light (PL) containing red, blue, and far-red components, or white light (WL) at a color temperature of 6500 K. Sorrel responses differed markedly among spectral environments and could not be explained by a single growth parameter. WL produced the highest mean shoot dry weight (505 mg), which was approximately 30.5% higher than that under PL (387 mg), although the difference between the two treatments was not statistically significant. WL also produced 46.5% greater leaf area than PL (50.1 versus 34.2 cm2) and showed the highest mean PIABS value (4.00). In contrast, PL showed the highest mean root dry weight (25.5 mg) and Dickson quality index (9.4), together with a relatively low shoot-to-root ratio (17.7) and top-heavy index (20.0), indicating a less shoot-biased structural pattern. BL restricted growth and leaf development and was associated with lower soil plant analysis development units, normalized difference vegetation index, photochemical reflectance index, Fv/Fm, and PIABS, indicating that BL alone may have limited suitability for sorrel cultivation under the present experimental conditions. Under the tested conditions, WL appeared more suitable for yield-oriented production, whereas PL may be considered when shoot–root structural balance is prioritized.

International Journal of Plant Biology doi: 10.3390/ijpb17080063

Authors: Lucía Vásquez-Hernández Galo Pabón-Garcés

Opuntia ficus-indica (L.) Mill. is a crassulacean acid metabolism (CAM) xerophyte whose intraspecific morphological diversity remains poorly documented in Andean agroecosystems. This study characterized the phenotypic diversity of 55 accessions collected along an altitudinal gradient in the dry inter-Andean valleys of northern Ecuador. Twenty morphological descriptors were used to analyse the data. Quantitative descriptors showed moderate to high variability, with reproductive and defensive characters exhibiting greater variation than vegetative descriptors. MCA revealed substantial chromatic diversity, with fruit peel and pulp color emerging as the strongest qualitative discriminators among morphotypes. Cluster analysis identified three statistically distinct groups: a small-fruited, highly spinescent morphotype restricted to the most arid sites; an intermediate and phenotypically diverse morphotype distributed across all provinces; and a large-fruited, low-spinescence morphotype with the highest seed numbers, consistent with a more advanced domestication trajectory. The primary differentiation axis reflected a trade-off between spine investment and reproductive output. Five discriminant descriptors—fruit weight, spine length, seed number, peel color, and pulp color—provide a robust baseline for germplasm characterization and support conservation, breeding, and nutraceutical valorization in Andean drylands. The findings of this research contribute to filling a critical knowledge gap regarding Andean cactus pear germplasm, providing a scientific basis for conservation, sustainable management, and future breeding and agro-industrial initiatives.

International Journal of Plant Biology doi: 10.3390/ijpb17080062

Authors: Risky Mulana Anur Muslimah Arniyanti Intan Ria Neliana Bambang Sugiharto Wahyu Indra Duwi Fanata Tri Handoyo Parawita Dewanti

Water deficit is one of the most critical factors for determining the growth and yield of sugarcane. Understanding the physiological and molecular mechanisms of sugarcane responses is essential for developing resilient cultivars. In this study, three sugarcane cultivars, NX04 (sensitive), BL (moderate), and NXI-4T (tolerant), were grown in a greenhouse for 2 months and then subjected to drought stress for 8 days after planting. Morphological variation showed that the tolerant sugarcane cultivar exhibits a longer root system and delays leaf chlorosis and rolling. Malondialdehyde (MDA) content was increased in the sensitive and moderate cultivars, although it slightly increased in the tolerant cultivars at 8 days after drought stress. The increase was accompanied by increases in proline content and in gene expression of the catalase (Cat) and ascorbate peroxidase (Apx) across all cultivars, which protect cells from oxidative damage. Interestingly, the expression of the photosynthetic Pepc (phosphoenolpyruvate carboxylase) and Sps (sucrose-phosphate synthase) genes was significantly decreased, whereas SPS activity increased under drought stress. This implies that the SPS protein may be regulated through post-translational modification. The expression of transcription factors (TFs) of NAC, rather than DREB, was significantly upregulated in the tolerant cultivar under 8 days of drought stress, in line with the delay of chlorosis.

International Journal of Plant Biology doi: 10.3390/ijpb17080061

Authors: Safouane Benjaa Rachid Bouharroud Salahddine Chafiki Soumaya El Assri Abdelmalek Mahroug Ahmed Wifaya My Abdelmajid Kassem Redouan Qessaoui

Pathogenic strains of Fusarium oxysporum are major soilborne fungal pathogens responsible for Fusarium wilt in tomato, leading to significant yield losses worldwide. This study evaluated the biocontrol potential of rhizospheric bacterial isolates from argan (Argania spinosa) and raspberry (Rubus idaeus) soils through an integrated approach combining in vitro screening, greenhouse validation, and phylogenetic analysis. A total of 27 bacterial isolates were screened for antifungal activity using dual culture assays, of which ten exhibited more than 50% inhibition of fungal growth. Selected isolates were further evaluated for volatile organic compound (VOC)-mediated inhibition. Despite strong in vitro performance for several isolates, greenhouse experiments revealed that antifungal activity in vitro was not a reliable predictor of in planta efficacy. Among the tested isolates, BSA25, BSA23, and BSF8 significantly reduced disease severity and incidence under greenhouse conditions, with BSA25 achieving the greatest suppression. In addition to disease control, certain isolates promoted plant growth under pathogen stress, indicating dual functionality as plant growth-promoting rhizobacteria (PGPR). Molecular identification based on 16S rRNA gene sequencing and phylogenetic analysis (Neighbor-Joining, Kimura 2-parameter) revealed that the isolates belong to PGPR-associated genera, including Bacillus and Pseudomonas, while also highlighting functional variability among closely related taxa. Overall, this study demonstrates that multi-trait evaluation, integrating mechanistic screening and in planta validation, provides a more reliable framework for selecting effective biocontrol agents. The identified isolates, particularly BSA25, represent promising candidates for further evaluation for sustainable management of Fusarium wilt in tomato production systems.

International Journal of Plant Biology doi: 10.3390/ijpb17080060

Authors: Mariam Kiebre Jacques Ouedraogo Ignace Tondé Jocelyne Wendpingrenoma Ouangraoua Clémence Zerbo Zakaria Kiebre Romaric Kiswendsida Nanema Fanta Reine Sheirita Tietiambou Hamid El Bilali Filippo Acasto Jacques Nanema

Effective utilization of plant genetic resources requires agro-morphological evaluation of collections across diverse growing regions. The objectives of this study are to assess genetic diversity and estimate genetic parameters through morphological traits of Bambara groundnut. The study was conducted over two growing seasons at the SUSTLIVES project site in Gampela (Central Burkina Faso) using an RCBD with three replicates. A total of eight qualitative and eight quantitative traits were used to estimate variability within a collection of 153 landraces. The results revealed a substantial variability in all the qualitative traits, with the number of modalities ranging from 2 to 10, seed color being the variable trait. Based on qualitative traits related to seed, the landraces were grouped into 12 morpho-groups. The analysis of variance showed the presence of significant variation between landraces and across years and their interactions for all of the quantitative traits. The heritability values ranged from 46.12% (seed weight per plant) to 88.66% (days to 90% maturity). The landraces KKou4, KKou2, ODap3, ODap4, KLay68, BPle2, KTog6, KSag11, ZMag4 and ODap1 were identified as high-yielding landraces. Future selection efforts for Bambara groundnut should therefore continue to evaluate the genotypes that have a high potential for grain yield under various pedoclimatic environments of the country.

International Journal of Plant Biology doi: 10.3390/ijpb17070059

Authors: Akzhunis Imanbayeva Nurzhaugan Duisenova Nazerke Tolep Aidyn Orazov Ainur Tuyakova Akimzhan Lukmanov

The reliable identification of cultivated and naturalised mulberries is complicated by morphological plasticity, the historical movement of the planting material, and partly discordant nuclear and plastid signals. We evaluated four field-identified Morus alba L. trees, one from each of four localities in Mangystau, Western Kazakhstan, using archived consensus sequences for ITS, matK, rbcL, and trnH-psbA, together with a low-coverage Oxford Nanopore Technologies (ONT) dataset. Because the design comprised one tree per locality (n = 4), analyses were restricted to accession-level descriptive comparisons, and no population-genetic, phylogeographic, or formal phylogenetic inference was attempted. Archived alignment summaries indicated mean pairwise distances of 0.13% for matK, 0.44% for ITS, 0.94% for rbcL, and 2.92% for trnH-psbA; these values are reported as retained dataset descriptors rather than estimates of population diversity. An additional product generated with Rosaceae-derived s6pdh primers was excluded because the target identity and orthology could not be verified. The ONT run yielded 19,958 pass reads (69.27 Mb; read N50 3574 bp). Reference-enriched assembly produced a 14,824 bp candidate plastid-associated contig, approximately 9.3% of a typical Morus plastome. Its short length, incomplete and non-collinear annotations, and the absence of retained depth, polishing, assembly graph, and join support diagnostics preclude its interpretation as a complete, circular, or structurally validated plastome. This study provides a transparent pilot baseline for Mangystau mulberries and establishes quality control criteria for replicated sampling, validated markers, and deeper organelle sequencing.

International Journal of Plant Biology doi: 10.3390/ijpb17070058

Authors: Md. Shahidul Islam Abdul Hannan Mohammad Anwar Hossain Md. Motiar Rohman Richard William Bell

Methylglyoxal (MG), a reactive carbonyl species, is now recognized as a novel signaling molecule regulating abiotic stress tolerance and plant growth. Using tolerant (BHL-25 and BHL-27) and susceptible (BARI Barley-6 and BHL-26) genotypes, the study aimed to reveal the detailed mechanisms of MG-priming-induced salt stress tolerance in barley (Hordeum vulgare L.). Seeds were primed with MG, and seven-day-old seedlings were transferred to hydroponic solution. After one week of seedling growth in hydroponic solutions, the salinity stress was imposed. The five treatments were as follows: control (0 salt + 0 mM MG), salt (16 dS m−1) + 0 mM MG, salt + 0.25 mM MG, salt + 0.5 mM MG, and salt + 1 mM MG. Significant genotype-dependent changes were observed in response to salt stress based on morphological, physiological and biochemical traits. Salt stress significantly impaired shoot length, shoot dry weight, root volume, root dry weight, relative water content, leaf Na+ content and K+/Na+ ratio. Salinity stress caused a significant increase in oxidative indices in all genotypes; however, the tolerant genotypes showed a lower increase. Importantly, in plants grown from MG-primed seeds, the negative effects of salt stress were reversed by modulating the K+/Na+ ratio, the activities of antioxidative and glyoxalase pathway enzymes as well as the redox state of ascorbate and glutathione. In conclusion, MG-mediated salt tolerance was linked to a reduction in reactive oxygen species (ROS) and activation of ROS and MG detoxification processes as well as favorable regulation of the K+/Na+ ratio, glutathione and ascorbate redox state that improved the growth of the barley plants under salinity.

International Journal of Plant Biology doi: 10.3390/ijpb17070057

Authors: Thayná Kelly Formiga de Medeiros Francisco Thiago Coelho Bezerra Gleyse Lopes Fernandes de Souza Marcos Vinícius da Silva Maria Beatriz Ferreira Dayane Gomes da Silva Daniela Rosário de Mello Robevania da Silva Alves Almeida Francisco Eudes da Silva Igor Eneas Cavalcante Alberício Pereira de Andrade Olaf Andreas Bakke Ivonete Alves Bakke Riselane de Lucena Alcântara Bruno

Rising salt concentration in water induces osmotic stress and ionic toxicity, compromising photosynthesis, growth, and survival of threatened species such as Paubrasilia echinata and highlighting the need for stress-mitigating compounds such as salicylic acid. This study evaluated the effect of salicylic acid on the morphophysiological responses of P. echinata seedlings irrigated with saline water. Treatments were arranged in a 2 × 3 factorial scheme, with two electrical conductivity levels of irrigation water (0.5 and 6.0 dS m−1) and three salicylic acid concentrations (0, 2, and 3 mM), randomized across 24 experimental plots containing five plants each in a randomized complete block design. Analyses included growth, relative water content, chlorophyll indices and fluorescence of seedlings. Irrigation water with an electrical conductivity of 6.0 dS m−1 reduced leaf number by 9.09% and PSII efficiency by 26.94%. Under saline conditions, salicylic acid at 2 and 3 mM increased root dry matter by 49.28% and 60.29%, respectively. The highest total chlorophyll index (79.03) was obtained at 2 mM, whereas leaf area peaked at 734.74 cm2 per plant at 3 mM. Salicylic acid enhances salt tolerance in P. echinata. At 2 mM, it improves seedling morphophysiological traits and may support restoration in salinized sites.

International Journal of Plant Biology doi: 10.3390/ijpb17070056

Authors: Behzad Heidari Dugassa Nemie-Feyissa Cathrine Lillo

Accurate chromosome segregation during meiosis requires protection of centromeric cohesion mediated by shugoshin (SGO) proteins and Protein Phosphatase 2A (PP2A). In Arabidopsis thaliana, genetic and cytological studies have demonstrated that PP2A B′ regulatory subunits are essential for maintaining centromeric cohesion; however, direct biochemical evidence for their interaction with SGO1 in plants is lacking. Here, we used microscale thermophoresis (MST) to investigate and quantify the binding between fluorescently labeled GST-SGO1 and the PP2A B′α and B′β subunits. SGO1 exhibited concentration-dependent binding to both subunits, with dissociation constants (Kd) of 5.49 ± 0.88 µM for B′α and 1.89 ± 0.38 µM for B′β, indicating an approximately threefold higher affinity for B′β. These results provide direct quantitative biochemical evidence for a specific interaction between Arabidopsis SGO1 and PP2A B′ subunits.

International Journal of Plant Biology doi: 10.3390/ijpb17070055

Authors: Griselida Rojas-Campos Raúl Vargas Anyela Marcela Ríos-Ríos Eyner Huaman Amilcar Valle-Lopez Manuel Oliva-Cruz

Physalis peruviana L. is an Andean crop of high nutritional and commercial value; however, the limited availability of uniform planting material restricts its large-scale propagation. Indole-3-butyric acid (IBA) is widely used to promote adventitious rooting, although its concentration and application methods influence the response observed throughout the different stages of root development. This study evaluated how IBA concentration and explant origin influenced rooting-related traits and early vegetative growth of in vitro-regenerated P. peruviana shoots during a 30-day ex vitro acclimatization phase. The evaluated variables included rooting percentage, root number, longest root length, root fresh and dry mass, shoot length, leaf and node number, stem diameter, shoot fresh and dry mass, leaf area, and photosynthetic pigment contents. The experiment followed a completely randomized design with a 2 × 4 factorial arrangement consisting of two explant origins (cotyledon and hypocotyl) and four IBA concentrations (0, 400, 800, and 1600 mg L−1), with five biological replicates per treatment combination. All shoots formed at least one visible root, resulting in 100% rooting across all treatment combinations. IBA concentration significantly affected root fresh and dry mass and several shoot-growth traits, whereas root number and longest root length were not significantly affected. Among the concentrations tested, 800 mg L−1 produced the highest root biomass and favorable responses in selected shoot-growth traits, whereas 1600 mg L−1 was associated with lower values for some growth variables. Hypocotyl-derived shoots had more leaves and nodes, greater stem diameter, and higher shoot dry mass than cotyledon-derived shoots. These results indicate a concentration- and trait-dependent response to IBA and identify 800 mg L−1 as the most favorable concentration among those tested for increasing root biomass and selected shoot-growth traits under the evaluated acclimatization conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17070054

Authors: Mst. Lailatul Ferdows Saima Biswas Mizanur Rahman F. M. Jamil Uddin Md. Nayan Aporna Tabassum Api Mohaiminul Islam Nadia Islam Swapan Kumar Paul Md. Harun Rashid

A field experiment was conducted at the Agronomy Field Laboratory of Bangladesh Agricultural University to evaluate the effects of arbuscular mycorrhizal fungi (AMF) inoculation and foliar supplementation of nitrogen (N) and phosphorus (P) on the performance of maize (Zea mays L.) under irrigated and rainfed conditions. The experiment followed a randomised complete block design with two levels of AMF (inoculated and non-inoculated) and four foliar treatments (no N and P, N only, P only, and combined N + P). The recommended dose of fertiliser (RDF) was applied as a soil application to all treatments. AMF inoculation significantly increased grain yield by 8.1% under rainfed and 10.0% under irrigated conditions compared with non-inoculated plants. Foliar application of N and P, especially when they were applied together, significantly improved yield components, including cob length, number of grains per cob, and 1000-grain weight. The highest grain yield (9.08 t ha−1 under rainfed and 10.91 t ha−1 under irrigated conditions) was obtained from the combined treatment of AMF inoculation and foliar N + P application. Redundancy analysis (RDA) and linear mixed-effects modelling (LMM) confirmed that water availability was the dominant factor controlling maize productivity, while AMF inoculation exhibited a stronger independent effect than foliar fertilisation. Under rainfed conditions, the treatment responses were reduced, and maize responded better to foliar N application in combination with AMF than combined N + P application. These findings indicate that the integrated use of AMF and foliar nutrient management enhances maize productivity, but optimal combinations depend strongly on moisture regimes.

International Journal of Plant Biology doi: 10.3390/ijpb17070053

Authors: Silvana Marigorda-Castro Karol Vilchez-Estrada Javier Javier-Alva Yuliana Mendoza-Martínez Delia Talledo-Ancajima Krizia Pretell-Monzón Benoit Diringer Carlos Granda-Wong William Nauray-Huari Gastón Cruz

Neltuma pallida is a multi-purpose tree species of the seasonally dry tropical forests of northern Peru, where it provides essential ecological and socioeconomic functions. However, recurrent defoliation associated with the cecidomyiid gall midge Enallodiplosis discordis may compromise early seedling establishment and the success of forest restoration programs. This study evaluated the effects of larval infestation on foliar integrity and established quantitative damage thresholds in N. pallida seedlings under dry forest conditions. Insects collected from naturally infested plants were identified using an integrative taxonomic approach that combined classical morphological diagnosis with COI-based DNA barcoding obtained by Sanger sequencing. Morphological assessment assigned the defoliating dipteran to E. discordis, while BLASTn v2.17.0. analysis of the 576-bp partial COI sequence showed 92.6% identity and 100% query coverage with Cecidomyiidae records, supporting its taxonomic placement within this family. Field bioassays conducted over a 17-week period, in which 25 individual seedlings were evaluated (N = 25), revealed a strong and significant positive correlation between larval density and foliar damage percentage (r = 0.872; p < 0.001), with moderate damage levels predominating throughout the evaluation period. Despite sustained larval presence, seedlings did not reach severe damage categories, suggesting potential relative tolerance to partial defoliation under the evaluated field conditions. Temperature and relative humidity were not significantly associated with infestation intensity or foliar damage during the study period. Overall, these findings indicate that E. discordis-associated foliar damage represents a relevant, although not necessarily lethal, biotic constraint for the early regeneration of N. pallida under the field conditions assessed. The quantitative thresholds reported here provide useful criteria for dry forest restoration programs, phytosanitary monitoring, and integrated pest management strategies in the Peruvian dry forest.

International Journal of Plant Biology doi: 10.3390/ijpb17070052

Authors: Glynn C. Percival

The frequency and intensity of heatwaves are increasing annually worldwide due to climate change. Combined with the urban heat island effect, elevated heat stress episodes threaten the survival and performance of urban trees, in turn reducing their ecosystem benefits. For this reason, the foliar heat tolerance of 35 Malus genotypes (two species, 32 cultivars, one variety, one hybrid) was evaluated under controlled laboratory assays. Heat injury to foliar tissue was quantified using chlorophyll fluorescence (Fv/Fm) to assess photosystem II (PSII) damage and an electrolyte leakage index (ELI) to evaluate cellular membrane integrity. A preliminary dose–response experiment using six genotypes exposed to a temperature gradient (40–50 °C) was conducted to establish thermal response curves and derive LT50 values (temperature at 50% decline in Fv/Fm). These analyses confirmed substantial genotypic variation in thermal tolerance and identified 45 °C as an optimal discriminatory temperature for large-scale screening. This temperature was subsequently applied to assess heat injury across all 35 genotypes. Measurements were conducted in May (spring foliage) and August (summer foliage) to evaluate ontogenetic influences. In some instances, only one genotype was available for experimental purposes. Consequently, conclusions regarding genotypic differences in heat tolerance are based on replicated datasets, whereas genotypes represented by single-tree sampling are presented for descriptive purposes only. Heat stress significantly affected Fv/Fm and ELI, with strong genotype and seasonal effects recorded. In most genotypes, foliar damage was greater in spring than in summer. Good correlations between Fv/Fm and ELI confirmed their value as complementary physiological measures of heat tolerance in plants. Of the 35 genotypes evaluated, Malus sargentii, M. ‘Prairifire’, M. baccata ‘Jackii’, M. ‘Royal Fountain Huber’ and M. Donald Wyman were the most heat tolerant. The substantial variation in foliar heat tolerance detected across the 35 genotypes tested demonstrates potential for selecting Malus genotypes with superior foliar heat tolerance and highlights opportunities for identifying heat resilient candidates among other under-utilized urban tree taxa.

International Journal of Plant Biology doi: 10.3390/ijpb17070051

Authors: Ichiro Tono Makoto Saito Fujio Terai Yoshiro Baba Hiroyasu Ishikawa

In response to the need for a simple, non-destructive method for evaluating avocado ripeness, we measured chlorophyll-related fluorescence and chromaticity of the outer skin using simple optical equipment and evaluated their relationship with whole-fruit compression (wfc), which was used as a firmness-based ripeness index. A compact system consisting of a blue LED excitation source and a small spectrometer was used to measure fluorescence spectra, and a commercially available colorimeter was used to evaluate chromaticity. Hass avocado samples purchased from multiple retail stores in Japan and stored for different periods were examined. The combination of the fluorescence intensity ratio I740/I685 and the lightness parameter L* showed a moderate correlation with wfc, with R2 = 0.48. The fluorescence ratio I740/I685 was treated not as a direct measure of chlorophyll content, but as a spectral index associated with ripening-related changes in avocado skin, including chlorophyll-related fluorescence and skin optical properties. These results suggest that the combination of simple blue LED-excited fluorescence and chromaticity measurements may be useful as a practical screening approach for roughly estimating avocado ripeness in commercially available fruit.

International Journal of Plant Biology doi: 10.3390/ijpb17070050

Authors: Mohamed Ouknin Youssef Karra Hasnaâ Harrak Abderraouf El Antari Omar Drissi Abdelghani Tahiri Ahmed Wifaya Fouad Elame Meriyem Koufan Redouan Qessaoui Rachid Bouharroud Naima Ait Aabd

Argan tree (Argania spinosa L. Skeels), an endemic Moroccan species, is widely recognized for its traditional medicinal and nutritional uses. It has long been employed to promote skin and cardiovascular health, regulate blood glucose levels, and support overall wellbeing. Traditionally, different parts of the argan tree, including argan oil, leaves, and other plant-derived preparations, have been used to manage various health conditions such as diabetes, gastritis, gastric ulcers, rheumatism, joint and muscle pain, skin disorders including acne, eczema, and inflammation, as well as wound healing and dental problems. This narrative critical review compiles and evaluates current knowledge on the ethnobotany, phytochemistry, pharmacology, and toxicology of argan tree to support its evidence-based application. Relevant literature was collected from major English and French scientific databases, focusing on studies addressing the plant and its principal bioactive constituents. Ethnobotanical data indicate the extensive use of argan oil, leaves, and other plant parts in traditional remedies and dietary practices. Phytochemical investigations reveal a rich composition dominated by unsaturated fatty acids, tocopherols, phytosterols, and polyphenolic compounds. Experimental studies highlight a broad spectrum of biological activities, including antioxidant, antidiabetic, antibacterial, and anti-obesity effects, along with emerging applications in nanotechnology. Toxicological findings generally suggest low toxicity and good safety profiles under tested conditions. Overall, A. spinosa exhibits substantial ethnopharmacological relevance and diverse bioactivities, supporting its continued exploration for nutraceutical and therapeutic applications.

International Journal of Plant Biology doi: 10.3390/ijpb17060049

Authors: Klaudia Lukáčová Vanda Assunta Prota Miroslav Habán Grazia Maria Scarpa

Rosa damascena Mill. is a medicinal and aromatic species of major pharmacological and economic importance, widely valued for its complex profile of bioactive secondary metabolites. While extensive research has focused on field-grown plants and essential oils, comparatively little attention has been devoted to the behavior of R. damascena under in vitro conditions. Plant tissue culture systems provide controlled platforms for investigating secondary metabolism independently of environmental variability; however, their application to R. damascena has produced heterogeneous and often inconsistent results. This review examines the main in vitro culture systems developed for R. damascena, including callus, suspension, and organ-derived cultures, with emphasis on their capacity to accumulate secondary metabolites. Available evidence indicates that undifferentiated cultures generally fail to reproduce the full metabolic complexity observed in planta, particularly for volatile monoterpenes associated with tissue specialization. Nevertheless, several studies demonstrate that in vitro systems can accumulate phenolic compounds with relevant biological activities, supporting their use as experimental models for investigating metabolic regulation. By integrating early studies with recent advances in plant biotechnology, this review highlights current limitations, unresolved questions, and future perspectives for the use of R. damascena in vitro cultures in medicinal plant research.

International Journal of Plant Biology doi: 10.3390/ijpb17060048

Authors: Aigul Madenova Raigul Abdikarimova Zhankeldy Aitymbet Moldir Askarova Zarina Yussupova Irina Kovalchuk Svetlana Dolgikh Aigerim Seisenova Dinara Kaldybayeva Marina Urazaeva Sagi Soltanbekov Balnur Kabylbekova

Apple (Malus domestica Borkh.) rootstocks play a key role in modern intensive orchard systems, where their accurate identification is essential for breeding, nursery production, and certification of planting material. This is particularly important in Kazakhstan, a recognized center of origin of cultivated apple, where local germplasm remains insufficiently characterized at the molecular level. In this study, we integrated simple sequence repeat (SSR) genotyping and morphological trait analysis to develop a reliable approach for the identification of clonal apple rootstocks cultivated in Kazakhstan. Five widely used rootstocks (Zhetysu 5, ARM-18, B-7-35, M9, and B9) were analyzed using 17 polymorphic SSR markers and 30 vegetative traits. SSR analysis revealed moderate genetic polymorphism (PIC = 0.28–0.54; He = 0.35–0.58) and enabled clear discrimination among all studied genotypes. Cluster analysis based on genetic distances grouped rootstocks according to their genetic similarity, reflecting their origin and differentiation. Morphological evaluation demonstrated significant phenotypic variability and identified correlations among key vegetative traits related to plant vigor and leaf development. The integration of molecular and morphological data allowed the development of comprehensive genotype profiles (“molecular–morphological passports”) for each rootstock, ensuring their reliable identification. The proposed approach provides a practical framework for the certification of planting material and the management of apple genetic resources in Kazakhstan. It can be applied to improve nursery systems, support breeding programs, and ensure the production of true-to-type planting material in modern horticulture.

International Journal of Plant Biology doi: 10.3390/ijpb17060047

Authors: Joo Young Kim Karina Idiyatullina Thomas A. Colquhoun

Viola odorata, commonly known as sweet violet, is valued for both its fragrance and medicinal properties. However, seeds of V. odorata exhibit non-deep physiological dormancy, resulting in poor and inconsistent germination. This dormancy can be overcome through physical or chemical treatments, including scarification, stratification, and hormone application. Although mechanical scarification is effective, many commonly used approaches have notable limitations, such as reliance on corrosive chemicals and a lack of uniformity. This study presents a simple and effective mechanical scarification technique using rat-tooth tweezers to gently crack the seed coat tip of V. odorata ‘Empress Augusta’ (EA). This method significantly improved germination. When combined with cold stratification at 4 °C, germination further increased, reaching 70% within 8 weeks. Germination was enhanced even further on Murashige and Skoog (MS) basal salt medium supplemented with 10 mg/L gibberellic acid (GA3), achieving 97.5% germination by day 54. These findings suggest that this simple mechanical scarification method, when combined with cold stratification and GA3 treatment, could provide a reliable and practical strategy for breaking dormancy and facilitating seed germination in V. odorata.

International Journal of Plant Biology doi: 10.3390/ijpb17060046

Authors: Fernando Gomes Hoste Cristhiane Tatagiba Franco Brandão Ana Júlia Câmara Jeveaux-Machado Janyne Soares Braga Pires Felipe Moro Daniel Carvalho de Araújo Bliane Morozini Bacheti Maria Ester Lenzi de Souza Danilo Balla Filho Vinicius de Souza Oliveira Enilton Nascimento de Santana Karin Tesch Kuhlcamp Lúcio de Oliveira Arantes Sara Dousseau-Arantes

Protein hydrolysate-based biostimulants have been increasingly investigated due to their potential to improve seedling growth and physiological responses; however, integrated interpretations of dose-dependent morphophysiological and photochemical responses in Theobroma cacao L. during nursery production remain limited. This study evaluated the effects of increasing concentrations of Terrativa®, an organomineral biostimulant formulated with animal-derived protein hydrolysate, on growth, chlorophyll indices, and photosystem II performance in cacao seedlings of the Catongo and TSH1188 genotypes. The experiment was conducted in a randomized block design in a 2 × 6 factorial arrangement, corresponding to two genotypes and six Terrativa® concentrations (0, 1, 2, 3, 4, and 6 mL L−1). Most morphological and physiological variables exhibited quadratic responses. Intermediate concentrations promoted greater vegetative growth and improved photochemical responses, with maximum values observed for number of leaves (13.98 at 2.50 mL L−1), leaf area (994.10 cm2 at 2.91 mL L−1), stem length (42.79 cm at 3.44 mL L−1), root volume (9.21 cm3 at 3.70 mL L−1), and total chlorophyll (34.58 at 3.08 mL L−1). Catongo seedlings showed greater chlorophyll accumulation, whereas TSH1188 exhibited higher ABS/RC, TRO/RC, and PIabs values, indicating genotype-dependent differences in photochemical energy utilization. Higher concentrations were associated with reduced performance in several variables, suggesting lower physiological efficiency under elevated biostimulant doses. Overall, Terrativa® concentrations between 2.50 and 3.70 mL L−1 were the most effective for promoting balanced seedling development in both cacao genotypes, with Catongo responding more strongly in chlorophyll accumulation and TSH1188 showing greater photochemical responsiveness.

International Journal of Plant Biology doi: 10.3390/ijpb17060045

Authors: Muhammad Fikri Muh Farid Muhammad Fuad Anshori Amin Nur Nirwansyah Amier Salwa Aulia Haruni

The development of new corn varieties is necessary to meet the corn demand. Using full diallel crosses is a method for developing high-yielding hybrid corn. This development requires systematic selection methods that incorporate various approaches in developing selection indices. This study aimed to develop a selection index concept for two full diallel cross populations and select potential hybrid crosses for preliminary yield evaluation. The study involved two populations of 100 corn seed genotypes from full diallel crosses (90 F1 genotypes and 10 selfing elders) and five check varieties per population, planted using a Type II Augmented RCBD in eight blocks. Agronomic characteristics were analyzed using analysis of variance, heritability, factor analysis, and path analysis, with selection criteria aligned with heterotic potential, specific combining analysis, and heterobeltiosis. Analysis revealed significant genetic variation and moderate-to-high heritability for most traits. Correlation, factor, and path analyses identified cob diameter, number of rows per cob, and seeds per row as optimal selection criteria. Selection indices were developed by integrating standard heterosis, specific combining ability, and heterobeltiosis, with weights based on heritability and direct effects. Forty-four hybrid crosses showed potential for preliminary yield tests, with seven having the best final index compared to the reference variety. The p17 × p23 cross had the best potential for the final index. This study demonstrates the effectiveness of integrating multivariate analysis and selection indices in developing superior hybrid corn crosses. Further optimization is recommended through preliminary yield tests and molecular approaches.

International Journal of Plant Biology doi: 10.3390/ijpb17060044

Authors: Mini Devi Vasudha Datta Satish Kumar Bunty Sharma Damandeep Kaur Hardeep Singh Tuli Shafiul Haque Diwakar Aggarwal

Epigenetic regulation plays a significant role in controlling plant secondary metabolism and offers a promising strategy to enhance medicinal alkaloid production without altering the genomic sequence. This review highlights major plant epigenetic mechanisms, including DNA methylation, histone modifications, chromatin remodeling, and non-coding RNAs, and their influence on alkaloid biosynthetic pathways. Particular emphasis is placed on chemical epigenetic modulators, such as DNA methyltransferase and histone deacetylase inhibitors, which can activate silent metabolic genes in a rapid, reversible, and non-GMO manner. Evidence from medicinal plants demonstrates tissue-, dose-, and time-dependent enhancement of alkaloid production. The review further discusses emerging approaches including CRISPR-based epigenome editing, multi-omics integration, and advanced bioinformatics for systems-level understanding of epigenetic regulation. Despite challenges, such as limited species-specific datasets, and concerns regarding long-term stability, epigenetic modulation represents a promising tool for sustainable alkaloid production with important pharmaceutical and industrial applications.

International Journal of Plant Biology doi: 10.3390/ijpb17060043

Authors: Fernando H. Calderon-Quispe Francisco Medina Castro Salvador Mamani Cosi

Puya raimondii, an endangered high-Andean species, exhibits life-history traits that may strongly influence its reproductive strategy; however, its reproductive biology remains poorly understood. Here, we assessed its floral biology, breeding system, and pollination ecology through field observations and controlled pollination experiments. Flowers are incompletely protogynous and herkogamous, with subsequent androecium development leading to contact between anthers and receptive stigma, enabling autonomous self-pollination. Consistent with this mechanism, P. raimondii was found to be self-compatible and capable of autonomous reproduction. The fruit set was high in both populations studied, likely supported by extended floral longevity, abundant nectar and pollen rewards, and a generalized bird pollination system. Hummingbirds were the most frequent visitors; however, passerine birds, including Diglossa brunneiventris, as well as bees, also contributed to pollination to a different extent. These findings indicate that P. raimondii combines reproductive assurance through selfing with the ecological benefits of generalized pollination. This dual strategy may enhance reproductive success under the variable and pollinator-limited conditions of high-Andean ecosystems, highlighting the importance of further ecological and reproductive studies to inform conservation efforts for this endangered species.

International Journal of Plant Biology doi: 10.3390/ijpb17060042

Authors: Ying Wang Liner Zhuo Jinyi Wu Xiaotong Wang Hengfei Lv Xinmin Huang Qinqin He

Verticillium wilt caused by Verticillium dahliae poses a severe threat to pepper (Capsicum annuum) production worldwide. Kunitz trypsin inhibitors (KTIs) play crucial roles in plant disease resistance, yet research on the CaKTI gene family in pepper, especially regarding its regulatory functions in resistance to V. dahliae, remains limited. In this study, members of the CaKTI gene family were systematically identified in the pepper genome, followed by comprehensive analyses of their physicochemical properties, phylogeny, chromosomal localization, conserved motifs, cis-acting elements in promoters, and expression profiles. A total of 22 CaKTI genes were identified, all harboring the beta-trefoil_STI superfamily domain. They were unevenly distributed across four chromosomes, with evident tandem duplication events, and exhibited tissue-specific and developmental stage-specific expression patterns. In the Verticillium-resistant pepper cultivar, five candidate CaKTI genes (CaKTI9, CaKTI6, CaKTI17, CaKTI18, and CaKTI22) were significantly induced and upregulated, particularly in roots, and their expression might be modulated by the methyl jasmonate signaling pathway. This study reveals the molecular features, evolutionary conservation, and defense-associated expression patterns of CaKTI genes in pepper and provides a preliminary exploratory basis for future research on disease resistance and molecular breeding.

International Journal of Plant Biology doi: 10.3390/ijpb17060041

Authors: Edinson Pooll Acuña-Ramirez Raúl Vargas Eyner Huaman Manuel Oliva-Cruz

Physalis peruviana exhibits highly variable and poorly reproducible morphogenic responses under in vitro conditions, limiting the development of reliable regeneration systems. This study evaluated how the interaction between thidiazuron (TDZ), naphthaleneacetic acid (NAA), and explant type influences shoot regeneration in the Colombiano ecotype of Physalis peruviana. A factorial design (2 × 3 × 2) revealed that morphogenic responses were primarily driven by interaction effects rather than by individual plant growth regulators. Hypocotyl explants cultured in 4.54 µM TDZ combined with 0.6 µM NAA showed the highest shoot production, shoot formation capacity, and elongation. In contrast, TDZ alone induced limited shoot production, did not support efficient and organized shoot development, and was associated with abnormal morphologies. The response to NAA was non-linear, with intermediate concentrations maximizing shoot regeneration response, indicating that morphogenic competence operates within a narrow hormonal range. Overall, regeneration in the Colombiano ecotype of P. peruviana evaluated here was governed by the interaction between hormonal balance and explant type, identifying favorable conditions for shoot regeneration under the conditions tested.

International Journal of Plant Biology doi: 10.3390/ijpb17050040

Authors: Vijay Rupa Naser A. Anjum Asim Masood Nafees A. Khan

Soil salinity as a major abiotic stressor has significantly affected crop production worldwide. However, plants have developed complex signaling networks that enable them to adapt and cope with such environmental shifts. Recent research has demonstrated the involvement of hydrogen sulfide (H2S) in signaling cascades that link plant development with stress tolerance management. Similarly, glutathione (GSH), a non-enzyme antioxidant, and a vital tripeptide, has been found to protect plants from oxidative damage and regulate metabolic functions under abiotic stress. As a potential scavenger of ROS, GSH maintains cellular redox homeostasis through the ascorbate-GSH cycle and acts as a signaling molecule for the sulfur-status of plants. This review focusses on: (i) revisiting the concept and current status of soil salinity; (ii) highlighting its impact at cellular and whole-plant levels; (iii) elucidating the role of a H2S and GSH in plant salt stress tolerance; and (iv) exploring the potential interactive roles of H2S and GSH in mitigating salinity impacts. This review will provide valuable insights into the complex network involving H2S and GSH, suggesting pathways for developing climate-resilient crops.

International Journal of Plant Biology doi: 10.3390/ijpb17050039

Authors: Anna Paula Marques Cardoso Walter Esfrain Pereira Juliane Maciel Henschel Diego Silva Batista Francisco Thiago Coelho Bezerra Maria Alaíne da Cunha Lima Gleyse Lopes Fernandes de Souza Patricia da Assunção Macedo Thayná Kelly Formiga de Medeiros Adailson Túlio dos Santos Silva Edmilson Gomes das Neves Magaly Morgana Lopes da Costa Aline Daniele da Cunha Lima Ewerton da Silva Barbosa Francisca Iris da Silva Souza

The availability of water is a limiting factor for the growth and productivity of yellow passion fruit (Passiflora edulis Sims). The use of bioregulators has been investigated as a strategy to mitigate the effects of abiotic stress. Different concentrations of SNP were evaluated on growth, gas exchange, photosynthetic pigments, chlorophyll fluorescence, and enzymatic activity in Passiflora edulis seedlings under different water conditions. The experiment was conducted in a randomized block design, in a 2 × 4 factorial scheme, with two irrigation conditions (80 and 30% of field capacity), combined with three concentrations of SNP (50, 100 and 250 µM) and water (control), with five replications. Water deficit reduced morphological, physiological, and enzymatic parameters. The application of SNP increased root fresh mass (23.56 g at the 100 µM dose) and leaf dry mass (8.21 g at 250 µM SNP), with increases of 24.52% and 30.52% compared to the values obtained under the 50 µM dose, respectively. The highest number of leaves (14) and leaf area (1183.3 cm2) was observed at 250 µM SNP, corresponding to increases of 7.70% and 17.27%, respectively, compared to plants without SNP application. Water deficit reduced growth, gas exchange, chlorophyll fluorescence, and enzymatic activity. SNP promotes improvements in growth; however, it does not mitigate water deficit effects in Passiflora edulis seedlings.

International Journal of Plant Biology doi: 10.3390/ijpb17050038

Authors: Lin Gou Shang Shi Chenghao Zhu Ling Liu Jianmin Tang Rong Zou Xiao Wei

Champereia manillana (Bl.) Merr. var. longistaminea is an evergreen small tree. It belongs to the genus Champereia Griff. (Opiliaceae), and its tender leaves or flower buds can be eaten. It also has important medicinal and nutritional values. Wild populations of C. manillana are small and has a phenomenon of deforestation. Market development is hindered by propagation constraints, including low seed germination rates and poor rooting of cuttings. Standardized cultivation protocols are currently lacking. This paper systematically reviews the current status of propagation and cultivation research on C. manillana and analyzed the primary challenges. Recent research indicated that seed germination obstacles had been preliminarily overcome, and 50% shading was identified as the optimal cultivation condition. However, challenges remain, including slow growth, lack of standardized water and fertilizer management, and unclear molecular mechanisms regulating development. Future research should focus on improving vegetative propagation efficiency, elucidating growth mechanisms via multi-omics, and establishing standardized cultivation protocols from breeding to harvest. These strategies are essential for the sustainable utilization of C. manillana resources.

International Journal of Plant Biology doi: 10.3390/ijpb17050037

Authors: Ma Isabel Reyes-Santamaria David Chávez-Trejo Aracely Hernández-Pérez René Velázquez-Jiménez Eliazar Aquino-Torres Amanulla Khan Antonio de Jesus Cenobio-Galindo Macario Vicente-Flores Iridiam Hernández-Soto

The jalapeño pepper (Capsicum annuum L.) is a crop of great economic and nutritional importance worldwide; however, increasing yield and quality under conditions of reduced synthetic inputs remains a significant challenge, mainly due to restrictions in plant nutrition and stress response capacity; in this context, plant-based biostimulants, such as Ricinus communis extracts, are of particular interest due to their potential to modulate plant metabolism, promote growth, and favor the accumulation of bioactive compounds. In this study, the effect of a foliar-applied biostimulant derived from a methanolic extract of Ricinus communis L. on the physiological, agronomic, and biochemical parameters of jalapeño peppers was evaluated under open field conditions. A randomized complete design with five treatments was established: three extract concentrations (T50: 50 mg L−1, T75: 75 mg L−1, and T100: 100 mg L−1), a commercial biostimulant (Pepton 85/16 ®), and an absolute control. Significant differences (α ≤ 0.05) were observed between treatments T50, T75, and T100 with the application of castor bean and the absolute control in stem diameter, fruit number, yield, and polar and equatorial fruit diameter, as well as phenols, flavonoids, and antioxidant capacity (ABTS and DPPH). The application of R. communis extract (T50, T75, and T100) significantly improved plant performance compared to the control, particularly in yield (up to 270%), fruit number (73%), shoot biomass (up to 38%), and root development (up to 32%). Furthermore, increases in chlorophyll content and in antioxidant-related compounds were observed, including phenols, flavonoids, ABTS, and DPPH (up to 17%). Spearman correlation analysis revealed strong associations between structural and metabolic variables, highlighting the relationship between stem diameter, fruit traits, and bioactive compound accumulation, as well as the link between chlorophyll content and reproductive performance. The 1H NMR analysis indicated the presence of secondary metabolites such as ricin, unsaturated fatty acids, and phenolic compounds; however, their isolation and relationship with the biostimulant activity of the extract require further specific studies. Overall, foliar application of R. communis extract improved the growth, productivity, and biochemical attributes of jalapeño pepper, highlighting its potential as a sustainable alternative for crop management.

International Journal of Plant Biology doi: 10.3390/ijpb17050036

Authors: Anawat Padpaiboon Nuttha Sanevas Pornsawan Sutthinon

Epiphytic and leafless orchids possess specialized root structures and host diverse associated microorganisms, which may contribute to their adaptation to limited access to water, nutrients, and photosynthetic tissues. Microscopic, anatomical, and molecular analyses of aerial and substrate roots of Chiloschista lunifera (Rchb.f.) J.J.Sm. revealed consistent phototrophic microbial biofilms on the velamen surface. These biofilms comprised filamentous and unicellular cyanobacteria, singular bacterial cells, and green algae. Morphological characterisation identified cyanobacteria belonging to five taxonomic orders, which are Chroococcales, Chroococcidiopsidales, Nostocales, Leptolyngbyales, and Synechococcales. 16S rRNA amplicon sequencing confirmed cyanobacterial dominance, with Chroococcidiopsis thermalis PCC 7203 strongly prevalent in root wash samples (up to 99.99% relative abundance), while root homogenate samples harboured a more diverse assemblage including Phormidiaceae, Leptolyngbya, Scytonema, and Calothrix. In addition, a green alga from Watanabeales (Jaagichlorella sp.) was identified based on morphological characteristics. TEM showed diverse cyanobacterial forms and unicellular green algae with well-developed photosynthetic structures. Root anatomy differed between root types. Substrate roots exhibited an inverted kidney-shaped transverse profile, whereas aerial roots were circular, with differences in velamen distribution reflecting adaptation for water retention and substrate attachment. Branched root hairs occurred on substrate roots, while aerial roots possessed unbranched root hairs, indicating functional specialization. Autofluorescence analysis revealed lignified tissues and abundant cortical chloroplasts, suggesting a photosynthetic role of the roots that may compensate for leaflessness. These findings enhance our understanding of the anatomical and ultrastructural features of epiphytic leafless orchid roots and may support future conservation and propagation efforts.

International Journal of Plant Biology doi: 10.3390/ijpb17040035

Authors: Nigussie Kefelegn Solomon Benor Gizachew Haile Gidamo Asnake Fikre Ming Pei You Martin J. Barbetti

Yield is a complex trait influenced by multiple components and their genetic behavior. Therefore, this study was designed to understand the complex nature of yield by uncovering relationships among traits and estimating the genetic parameters of lentil germplasm. One hundred and ninety-two lentil (Lens culinaris) germplasm samples were evaluated at Jamma and Enewari (field experiment) and at Debre Birehan agricultural research center (pot experiment) in Amhara Region, Ethiopia, in 2024 and 2025. An alpha lattice design was used for both set of experiment, and data on 12 agronomic traits were collected. Genetic parameter estimations, correlation, path, principal component and cluster analyses were performed in R. The results revealed substantial phenotypic and genetic variation among the evaluated germplasm, with high broad-sense heritability for flowering, maturity, and seed size traits, and moderate heritability for seeds per plant, plant height, harvest index, and yield. Yield exhibited a strong phenotypic correlation with harvest index (r = 0.78 in field and r = 0.95 in pots), biomass (r = 0.77 in the field and r = 0.78 in pots), seeds per plant (r = 0.42 in the field and r = 0.60 in pots), and podding (r = 0.45 in the field and r = 0.69 in pots). Similarly, genotypic correlations were high with harvest index (r = 0.83 in the field and r = 0.96 in pots), biomass (r = 0.75 in the field and r = 0.80 in pots), seeds per plant (r = 0.0.59 in the field and r = 0.58 in pots), and podding (r = 0.39 in the field and r = 0.68 in pot), and both their direct and indirect effects on yield were significantly high. Therefore, indirect selection through traits such as pods per plant, harvest index, biomass, and seeds per plant would be more effective and reliable for improving seed yield than direct selection for yield itself as this is highly affected by environmental variations.

International Journal of Plant Biology doi: 10.3390/ijpb17040034

Authors: Pablo Santana Vial Niquisse José Alberto Emanoel Chequetto Wellington Castrillon Grélla Laís da Silva Magevski Militino Paiva Carrafa Edilson Romais Schmildt Deurimar Herênio Gonçalves Júnior Fábio Luiz Partelli

This study evaluated the morphological development of 23 Coffea canephora clones in Espírito Santo to identify materials with superior vigor and quality for commercial and breeding purposes. Seedlings from cuttings were arranged in a completely randomized design with ten replicates and assessed at the commercial dispatch stage. Shoot and root growth, biomass, leaf area (LA), Dickson Quality Index (DQI), structural ratios (shoot/root ratio, SRR; height/diameter ratio, HDR), and anatomical traits were measured. Data were analyzed using analysis of variance with Scott–Knott clustering, Pearson correlation, and Principal Component Analysis (PCA). Significant variability was observed among clones. Clones 88, VR3, 8, and LB33 showed the highest stem diameter (SD), total dry mass (TDM), LA, and DQI, with balanced shoot and root development. Leaf area correlated strongly with SD, number of leaves (NL), biomass, and DQI, confirming its role as a seedling quality indicator. PCA identified two groups: a high-performance group with greater vigor and biomass, and a lower-performance group including clones 7, MR04, and VR4. The convergence of methods confirms the robustness of the results. Overall, clones 88, VR3, 8, and LB33 demonstrate superior agronomic potential at the seedling stage, offering promising options for nurseries, growers, and clonal selection programs.

International Journal of Plant Biology doi: 10.3390/ijpb17040033

Authors: Fatima Lakhdar Bouchra Benhniya Jamal Bouhraoua Selma Mabrouki Nazha Samri Badr-ddine El Mouns Taoufik Hachimi Nabila Boujaber Omar Assobhei Samira Etahiri

Phytopathogenic diseases are a major limiting factor in agricultural production. Therefore, scientific research continues to focus on developing effective techniques to mitigate their impact on crop productivity. Seaweed extracts, used as nutritional supplements, organic fertilizers, or bio-pesticides, have demonstrated their ability to enhance plant growth, increase yield, and alleviate the effects of abiotic stress. This study aimed to evaluate the effect of the aqueous extract of Bifurcaria bifurcata, collected from the Atlantic coast of Sidi Bouzid (El Jadida, Morocco), on the growth of Solanum tuberosum L., as well as its bactericidal activity against soft rot caused by Dickeya dadantii. The chemical Characterization revealed that Bifurcaria bifurcata aqueous extract is rich in polar and hydrophilic functional groups. In addition, this extract is particularly rich in phenolic metabolites, particularly phenolic acids, such as p-coumaric acid, ferulic acid, vanillic acid, and caffeic acid, which are known for their potential antimicrobial mechanisms. However, the treatment with 4 g/L extract resulted in a significant reduction in disease symptoms (>60%) and enhanced plant growth parameters, including 21% increase in plant height and 33% increase in leaf number. POX activity increased 6-fold (from 0.12 to 0.7 µmol/min/mg protein), indicating successful elicitation of plant defense mechanisms. The Bifurcaria bifurcata extract could act as novel activators of plant defense mechanisms and serve as potential alternatives to chemical pesticides.

International Journal of Plant Biology doi: 10.3390/ijpb17040032

Authors: Mingcan Fu Xianbin Liu Chengyu Zhang Jian Ding Bin Liu Xiangqian Wu Zhiyang Wang

Systematic comparisons of how plants with contrasting ecological strategies respond to extremely wide nutrient availability gradients remain limited. We investigated the physiological, photosynthetic, and growth adaptations of four plant species representing distinct ecological strategies: Triticum aestivum L. (C3 annual crop), Zea mays L. (C4 annual crop), Ipomoea aquatica Forssk. (C3 annual/perennial aquatic vegetable), and Canna glauca L. (C3 perennial wetland ornamental). Plants were grown hydroponically under nitrogen (N), phosphorus (P), and potassium (K) gradients ranging from 0% to 500% of standard Hoagland nutrient solution. The study results showed that all measured plant traits exhibited characteristic unimodal dose–response patterns. Optimal performance mostly occurred at 100–150% nutrient availability gradients. Severe inhibition or mortality occurred at extreme gradients. Simultaneously, different plant species displayed markedly varying response amplitudes and nutrient-specific sensitivities. Z. mays showed the highest nutrient use efficiency and broadest optimal ranges, particularly for N and K. C. glauca exhibited extraordinary N responsiveness (32-fold increase in photosynthetic rate) but narrow optimal ranges (e.g., 1.01 ± 0.15 μmol CO2/(m2·s) at the 1% N treatment vs. 32.52 ± 3.33 μmol CO2/(m2·s) at the 150% N treatment). I. aquatica showed pronounced P limitation with broad tolerance to supra-optimal N and K. T. aestivum displayed moderate responses with clear sensitivity to N limitation. Root–shoot ratios declined systematically with increasing nutrient availability across all plant species, following negative exponential functions. The results of data analyses revealed significant effects of N, P, and K availability on all the determined plant traits. Correlation analyses demonstrated tight coupling effects among physiological, photosynthetic, and growth traits, indicating integrated whole-plant responses to nutrient variations. These findings reveal that plant ecological strategy systematically modulates nutrient response patterns and provide a quantitative framework for species-specific nutrient management. This study provides a theoretical basis for precision fertilization of aquatic vegetables and wetland plants, and more broadly support species-specific nutrient management in controlled-environment agriculture.

International Journal of Plant Biology doi: 10.3390/ijpb17040031

Authors: Shitong Chang Wanlong Li Zhou Liu Xiaomei Zhou Xiaoxi Shen

Soybean (Glycine max L. Merr.) is one of the most important legume crops globally, providing high-quality plant protein and oil for humans and livestock, and playing a crucial role in nitrogen fixation within agricultural ecosystems. The seeds contain about 35–40% protein by dry weight, with 65–80% of this being seed storage proteins (SSPs). These proteins mainly consist of 11S globulin (glycinin) and 7S β-conglycinin, which accumulate significantly in protein bodies during seed development, providing essential nitrogen and amino acids for seed germination and early seedling growth. Additionally, the composition and structure of SSPs directly determine the nutritional value, processing functionalities (such as emulsification, gelation, and solubility), and potential allergenicity of soybean products. In this study, we conducted a detailed analysis of the structural characteristics, chromosomal localization, phylogenetic relationships, and tissue expression patterns of members of the soybean Gy gene family, laying a theoretical foundation for further exploration of the biological functions of Gy genes in soybeans. We performed comprehensive genomic identification, expression analysis, and subcellular localization of the soybean Gy gene family. The results showed that the seven soybean Gy genes are unevenly distributed across different chromosomes and exhibit distinct expression patterns in soybean seeds, suggesting they may have different roles during seed development. Subcellular localization experiments indicated that the GmGy1 gene might play an important role during seed development. These findings provide significant insights into the functions of the Gy gene family in soybean growth and development and offer potential candidate gene targets for soybean molecular breeding.

International Journal of Plant Biology doi: 10.3390/ijpb17040030

Authors: Khadija Khouya Houda Taimourya Soumia El Malahi Jamaâ Zim Ibtissam Lahrach Aya Elatrassi Bahija Zakri Abdellah Benbya Khadija Basaid Ouiam Lahlou Yasmina Imani Mounia Ennami

Climate change exacerbates water scarcity in semi-arid and arid regions, particularly across the Mediterranean Basin, posing severe challenges to food security and freshwater availability. Non-conventional water resources, such as desalinated seawater, are increasingly considered for supplementing irrigation; however, their exclusive use can induce osmotic stress, nutrient imbalances, and soil alkalinity, thereby limiting crop performance. This study evaluated the agronomic, and physiological impacts of blending freshwater (FW) and desalinated seawater (DSW) for two zucchini (Cucurbita pepo L.) cultivars, Radia and Kayssar, under greenhouse conditions. Five irrigation regimes were tested: T1 (FW100%), T2 (FW75%-DSW25%), T3 (FW50%-DSW50%), T4 (FW25%-DSW75%), and T5 (DSW100%). Moderate blending, particularly T2 and T3, optimized vegetative growth, biomass accumulation, and reproductive performance, maximum yields were obtained under T3, reaching 6.65 kg/plant for Radia and 5.49 kg/plant for Kayssar, while fruit quality, including caliber and soluble solids content (°Brix), was also highest under this regime. These findings support the suggestion that implementing such combined/blended irrigation regimes can enhance vegetative growth, yield, and fruit quality in the face of increasing water scarcity and energy constraints.

International Journal of Plant Biology doi: 10.3390/ijpb17040029

Authors: Abdul Rehaman Syed Nazar ul Islam Arif Tasleem Jan Sajid Khan Mohd Asgher Nafees A. Khan

The common bean (Phaseolus vulgaris L.cv. BR-104) is the most widely cultivated legume crop and serves as a major dietary protein source worldwide. However, climate change-induced drought poses a severe threat to its productivity by disrupting key physiological and biochemical processes. Therefore, identifying effective strategies to enhance drought resilience in the common bean is of considerable importance. The present study investigates the regulatory role of hydrogen sulfide (H2S) in improving drought tolerance. Polyethylene glycol (15% PEG) induced drought stress markedly reduced phenotypic changes (leaf area (LA), plant dry weight (PDW), root length (RL), and shoot length (SL) by 18.6, 20.5, 30.3 and 17.5% respectively), photosynthetic efficiency (Fv/Fm by 28.4%), and photosynthetic pigment concentrations (chlorophyll and carotenoids by 25.6 and 36%, respectively), while significantly elevating oxidative stress markers (H2O2 and TBARS by 137.1% and 169.8%, respectively), leading to impaired stomatal movement and damaged chloroplast structure. Exogenous H2S application as sodium hydrogen sulfide (200 µM NaHS; H2S donor) effectively alleviated drought-induced oxidative damage by boosting endogenous H2S and GSH levels, upregulating activity of antioxidative enzymes, SOD, APX, and GR, thereby promoting reactive oxygen species (ROS) scavenging, and minimizing lipid peroxidation. Moreover, H2S maintained photosynthetic efficiency via improved stomatal openings and chloroplast structure, thus sustaining chlorophyll levels and stabilizing photosystem-II functionality. Enhanced proline accumulation following NaHS application led to improved osmotic adjustment, thereby contributing to overall stress tolerance. The use of a H2S scavenger at 100 µM HT (Hypotaurine) suppressed the mitigating effects of H2S, confirming the role of H2S in enhancing drought tolerance in the common bean. Collectively, these findings highlight the potential effect of H2S as a regulatory signaling molecule to enhance drought resilience in the common bean under drought stress conditions. Further research should explore integrating H2S-based treatments with breeding programs and agronomic practices to develop sustainable strategies to improve drought resilience in legumes and other staple crops under changing climatic conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17040028

Authors: Andrey R. Suprun Stanislava A. Vinogradova Konstantin V. Kiselev Nikolay N. Nityagovsky Alexandra S. Dubrovina

Exogenously induced RNA interference (exoRNAi) is a powerful biotechnology tool for precise gene regulation. The plant chalcone synthase (CHS) gene serves as a valuable model for molecular biology due to its central role in flavonoid biosynthesis. However, there are currently very few studies addressing the advantages and disadvantages of in vitro (enzymatic) or in vivo (bacterial) methods for producing double-stranded RNA (dsRNA) for exogenous application. This study aims to optimize and compare the two methods for producing dsRNAs targeting the Arabidopsis thaliana CHS gene: enzymatic synthesis in vitro using a commercial kit and bacterial synthesis in vivo using an engineered E. coli HT115 (DE3) system. Bacterial synthesis conditions were optimized with respect to IPTG concentration and cultivation time, and the resulting dsRNA preparations were purified and quality-controlled. Their biological activities were assessed by treating A. thaliana plants and analyzing the effects on AtCHS gene expression and flavonoid production using qRT-PCR and HPLC-MS. The results demonstrated that purified AtCHS-dsRNA from both methods effectively suppressed AtCHS expression and downstream flavonoid biosynthetic gene expression, leading to significant reductions in anthocyanins and flavanols. This study confirmed the efficacy of exogenous dsRNAs in regulating plant metabolic pathways and provided a comparative analysis of dsRNA synthesis methods, highlighting their benefits and limitations for practical applications in plant biology and protection.

International Journal of Plant Biology doi: 10.3390/ijpb17040027

Authors: Eugenia León-Jiménez Federico A. Gutiérrez-Miceli Esaú Ruíz-Sánchez Daniel González-Mendoza Benjamín Valdez-Salas María C. Luján-Hidalgo Joaquín A. Montes-Molina Angel M. Herrera-Gorocica

The use of nanoparticles (NPs) synthesized from plant extracts is an alternative to conventional pesticides for the control of agricultural pests. This study aimed to optimize the conditions of synthesis of silver–zinc nanoparticles (Ag-ZnNPs) using extracts of Ocimum basilicum L. and Crotalaria longirostrata Hook. & Arn. and to evaluate their phytotoxic impact on maize seedlings. The Ag-ZnNPs (Ag-Zn nanoparticles) were synthesized by redox reaction between metal ions and reducing metabolites present in the extracts. A response surface methodology (RSM) with three factors (extract concentration, heating time and pressure) was applied to determine the optimal synthesis conditions. The phytotoxicity of nanoparticles (NPs) on maize seedlings was subsequently evaluated on root growth, oxidative stress enzymes (CAT, POD, and APX), and physiology of seedlings. Nanoparticles synthesized from C. longirostrata extract demonstrated superior properties, with an optimization of synthesis (R2 = 95.3%) where the extract concentration (1:4 v/v; p < 0.01) was the critical factor influencing the reduction of metallic ions to nanoparticles. These NPs exhibited superior stability, smaller size (<100 nm), and zeta potential greater than 30 mV compared with O. basilicum extracts. Their NPs exhibited poorer optimization of synthesis (R2 = 43.8%) without the effect of any of the variables evaluated. Essentially, C. longirostrata NPs showed no phytotoxic effects on maize seedlings’ physiological parameters and enhanced root growth (117.2 mm) without negatively affecting photosynthesis (PSII 70-81 FvFm). Ag-ZnNPs synthesized with C. longirostrata exhibited optimal stability and size, along with no observed possible phytotoxicity effects, unlike O. basilicum NPs, which cause stress on maize seedlings. Therefore, Crotalaria longirostrata NPs could represent a promising material for agricultural pest control, with no apparent adverse effect on maize crops.

International Journal of Plant Biology doi: 10.3390/ijpb17040026

Authors: Da Hyeon Lee Jungeun Cho Hyeon Park Tae Hyeon Heo Ju Kyong Lee

This study evaluated morphological variation in 45 accessions of cultivated Perilla frutescens var. frutescens (PFF) and var. crispa (PFC) collected from South Korea and Japan, together with their weedy counterparts, var. frutescens (WPFF) and var. crispa (WPFC) from South Korea, using ten quantitative and ten qualitative traits. Clear morphological differentiation was observed between cultivated and weedy forms in both varieties, particularly in pigmentation, plant fragrance, and seed-related traits. PFF cultivars were characterized by predominantly green pigmentation, a typical frutescens aroma, and significantly larger and heavier seeds, suggesting stronger phenotypic differentiation in leaf and seed utilization. In contrast, PFC cultivars showed substantial morphological overlap with WPFC and WPFF accessions, indicating comparatively weaker phenotypic differentiation. Principal component analysis showed that the first principal component (PC1) explained 25.7% of the total phenotypic variance and was strongly associated with five quantitative traits (plant height, inflorescence length, floret number, seed size, and 100-seed weight) and five qualitative traits (adaxial and abaxial leaf color, flower color, seed color, and seed hardness). Along PC1, PFF cultivars formed a well-defined cluster, whereas PFC cultivars and the weedy WPFF and WPFC accessions exhibited broader dispersion, reflecting greater morphological variability. These results provide morphological insights into the differentiation between cultivated and weedy Perilla accessions and indicate potential domestication-related patterns. However, these interpretations are based primarily on morphological observations, and further genetic and evolutionary studies will be necessary to clarify the domestication history of these taxa. The identified trait complexes provide a useful phenotypic foundation for marker-assisted breeding, informed cultivar selection, and effective germplasm conservation and management.

International Journal of Plant Biology doi: 10.3390/ijpb17040025

Authors: Ulyana Panova Olga Kotsupiy Evgeniya Karpova Elena Ambros

Astragalus glycyphyllos (Fabaceae) is known to be a source of flavones, flavonols, and isoflavones, and its in vitro culture may promote the efficiency and sustainability of obtaining pharmacologically valuable fractions. The aim of this study was to develop an effective plantlet regeneration protocol for A. glycyphyllos, providing the accumulation of phenolic compounds and antioxidants in cultured tissues. The results show a maximum seed germination rate (67.8%) after scarification (mechanical with sandpaper followed by treatment with 50% sulfuric acid) and subsequent sterilization with 1.1% sodium hypochlorite solution. The maximum regeneration rate (95%) was achieved on Murashige and Skoog medium supplemented with 0.5 mg·L−1 thidiazuron. A thidiazuron concentration of 0.05 mg·L−1, combined with a twofold increase in iron chelate content, induced the maximum yield of total flavonoids (8.74 mg·g−1 DW), and significant levels of total phenolics (4.15 mg·g−1) and antioxidants (1.83 mg AAE·g−1) in the microshoot tissues. HPLC analysis showed kaempferol glycosides (1.51 mg·g−1) and acylated kaempferol glycosides (2.76 mg·g−1) as major components. Formononetin in a modest amount (0.09 mg·g−1) was detected in hydrolyzed extracts. The phenolic profiles of the microshoots and native plants coincided in hydroxycinnamic acid composition; meanwhile, quercetin glycosides were present only in in situ plants, and formononetin was found only in the plantlets. The results confirm the prospects of biotechnological methods for the industrial production of standardized medicinal raw materials.

International Journal of Plant Biology doi: 10.3390/ijpb17040024

Authors: Patrick Chiza Chikoti Batiseba Tembo Xinyao He David Paul Hodson Aakash Chawade Pawan K. Singh

Wheat blast, caused by Magnaporthe oryzae pathotype Triticum (MoT), is an emerging fungal disease that poses a serious threat to global wheat production. In Zambia, where wheat is increasingly becoming a vital component for food and nutritional security, the emergence and spread of wheat blast is a growing concern under the influence of climate and agricultural practices changes. This review assesses the risk of wheat blast expansion in Zambia by examining regional climatic trends, future climate projections, crop suitability, and the ecological requirements of MoT. Potential disease hotspots are identified, and integrated management strategies, including chemical, cultural, and biotechnological approaches are evaluated. The review highlights the urgent need for coordinated disease surveillance, the development and deployment of resistant cultivars, and climate-resilient farming practices. By consolidating current knowledge and outlining sustainable management strategies, this paper aims to support effective disease mitigation and safeguard wheat production in Zambia in the face of climate change.

International Journal of Plant Biology doi: 10.3390/ijpb17030023

Authors: Cayetano Navarrete-Molina María A. Sariñana-Navarrete Cesar A. Meza-Herrera Ángeles De Santiago-Miramontes José L. Rodriguez-Alvarez Raúl A. Cuevas-Jacquez Luis M. Valenzuela-Núñez Ricardo I. Ramírez-Gottfried Edir Torres-Rodriguez Rubén I. Marín-Tinoco

The genus Juniperus species is widely distributed in the Northern Hemisphere of the planet Earth. These species are notable for their ability to adapt to extreme environmental conditions, playing a crucial role in ecosystem structure and function. Currently, their expansion is being driven by anthropogenic activities and climate change, posing significant challenges for both control and conservation. The objective of this review was to synthesize the available evidence regarding the ecological importance and impacts of Juniperus on ecosystems, promoting a holistic perspective that contributes to the achievement of the United Nations 2030 Agenda for Sustainable Development. A systematic literature search was conducted using the Scopus database, and only the documents published between 2001 and 2025 were considered for the investigation. The results showed that these species possess a high ecological versatility, favoring their invasive success in disturbed ecosystems, particularly under the influence of climate change and land-use changes. Conversely, Juniperus species facilitate positive ecological outcomes by providing essential ecosystem services that benefit both the human population and the flora and fauna present in these ecosystems. Nevertheless, their expansion also causes negative effects, such as the suppression of herbaceous shrubs and understory cover, alteration of the hydrological function, and accelerated soil erosion, among others. Consequently, the genus Juniperus exhibits a dual ecological role, acting as a hero to many species within these ecosystems, yet a villain to others. In this sense, given its remarkable adaptive dynamism under scenarios of climate change and continuous anthropogenic alterations, it is imperative to promote comprehensive conservation and restoration strategies. These should include ecological monitoring, invasive species control, genetic management, and habitat restoration. Such efforts must be supported by long-term interdisciplinary research to understand and mitigate the ecological, genetic, and social impacts resulting from its expansion. Furthermore, these investigations and strategies must be flexible and locally contextualized to promote genuine ecosystem resilience in the face of the ongoing environmental transformations.

International Journal of Plant Biology doi: 10.3390/ijpb17030022

Authors: Tatiane Otto de França Bárbara Martins Bruno Gonçalves de Oliveira Luiz Antonio Biasi Renato Vasconcelos Botelho António M. Jordão

The use of new grape cultivars is essential for the continued development of Brazilian viticulture. Thus, this study aimed to determine the general physicochemical parameters, global phenolic composition, and antioxidant capacity of grape musts from the Portuguese red variety ‘Touriga Nacional’ during ripening in two Brazilian vineyards (states of Rio Grande do Sul and Santa Catarina). The results were compared with data obtained from the same variety cultivated in a vineyard located in northern Portugal, which is the region of origin of this variety. This research was conducted over three consecutive vintages (2022–2024). Overall, the results indicated that soil and climate conditions at each location were associated with differences in the composition of ‘Touriga Nacional’ grape musts. Grapes from both Brazilian vineyards showed significantly higher berry weight, must volume, and yield compared with grapes collected from the Portuguese vineyard. On the other hand, grapes collected from the vineyard located in the state of Santa Catarina exhibited the highest values of total titratable acidity, malic acid, total phenols, flavonoids, total anthocyanins, and chromatic characteristics. Nevertheless, grapes collected from the Portuguese vineyard showed higher values of total tannins. The results suggest that the ‘Touriga Nacional’ variety shows better adaptation to the conditions of the Santa Catarina vineyard than to those of the Rio Grande do Sul vineyard. These findings help winegrowers, particularly in southern Brazil, to have more options for introducing different grape varieties, thereby contributing to the production of wines with distinctive characteristics, while consumers will have access to a greater diversity of wines available on the market.

International Journal of Plant Biology doi: 10.3390/ijpb17030021

Authors: Magdoline Mustafa Ahmed Osman Ronald Kuunya Rania Alrasheed András Tamás Illés Árpád Tamás Rátonyi

Water stress is a major challenge that limits the growth, development, and yield of maize (Zea mays L.) worldwide, especially under climate change, particularly abiotic stresses. This review presents a comprehensive bibliometric and literature-based analysis of research on maize’s response to drought and water scarcity from 1975 to 2025, using VOS viewer1.6.20 software, facilitating the detection of co-authorship networks, thematic groupings, and patterns of keyword co-occurrence within the selected publications. Data from the Web of Science were examined to assess publication trends, keyword networks, and international collaborations. A literature search was conducted by combining the keywords ((“maize”) OR (“corn”) AND (“drought”) OR (“water stress”) AND (“yield”)). Relevant studies were retrieved from the Web of Science (WoS) database using this search string. The Mann–Kendall test revealed a significant positive trend (p = 0.001) in publications on water scarcity (R2 = 0.8526), with 396 relevant studies identified globally, regardless of language. The analysis of publication trends demonstrated a statistically significant increase in the volume of publications over the examined period, featuring major contributions from Kenya, Switzerland, Mexico, China, and the United States. The most influential publication focuses on a biotic stressor that significantly reduces maize grain yield. These results emphasise the need for integrated strategies that combine genetic improvement and sustainable irrigation to mitigate the impacts of water stress. This comprehensive analysis provides a foundation for guiding future research and policy development to improve maize resilience against the effects of water stress under changing climatic conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17030020

Authors: Luis Cagua-Montaño Karen Rodas-Pazmiño Jorge Fabricio Guevara-Viejó Betty Pazmiño-Gómez Ignacio Isa-Vargas Samuel Valle-Asan Rodrigo Pazmiño-Pérez Stefany Pilar Jami Jami Ivana Alexandra Armijos Galarza Edgar Rodas-Neira Cristhian Emilio Delgado Espinoza

Seed germination and early root growth are decisive for crop establishment, yet responses to ionic environments can be strongly genotype-dependent. This study evaluated the effect of supplementing an agar-based in vitro system with a commercial NPK fertilizer on the germination dynamics and early seedling traits of Raphanus sativus L. Seeds were tested in two solid media: A (1.3% agar, no fertilizer) and AF (1.3% agar supplemented with 0.45 g of granular NPK fertilizer (15–30–15) per 200 mL medium), using a completely randomized 3 × 2 factorial design. Germination percentage and synchrony are key constituents of seedlot evaluation because they jointly capture both viability and the temporal coordination of emergence. However, final germination percentage alone does not reflect the timing and uniformity of germination, which can be critical for predicting establishment and subsequent performance. Therefore, indices such as mean germination time (MGT), coefficient of velocity of germination (CVG), and interval germination rates are frequently employed to describe germination dynamics. In addition to germination dynamics, early seedling morphometry (e.g., root and hypocotyl traits) can provide complementary information on early vigor and stress sensitivity under contrasting media or environmental conditions. Root elongation was significantly reduced by fertilization in ASD and GE, whereas AS exhibited consistently shorter roots with no significant response. PCA summarized 86.3% of the total variance in the first two components, separating treatments along a vigour/architecture axis and a germination capacity axis (%G), and hierarchical clustering identified five response groups. Overall, a low-cost agar + fertilizer system effectively discriminated genotype-specific sensitivity to an ionic environment during early establishment, highlighting the need to consider variety-dependent thresholds when using commercial fertilizers for in vitro screening.

International Journal of Plant Biology doi: 10.3390/ijpb17030019

Authors: Almagul Malimbayeva Batyrgali Amangaliev Erbol Zhusupbekov Akerke Soltanayeva Aina Sagimbayeva Zhuldyz Oshakbayeva Karlyga Rustemova Maksat Batyrbek

Efficient nutrient management is essential for enhancing flax productivity under semi-arid conditions. A two-year field experiment (2024–2025) was conducted in southeast Kazakhstan to evaluate the effects of integrated foliar fertilization with macro- and micronutrients and biofertilizers on the growth, seed quality, and yield of flax (Linum usitatissimum L.). The experiment was arranged in a randomized complete block design with five treatments: T1 (control, absolute zero fertilization (N0P0K0), i.e., soil without any additional nutrient applications), T2 (N60P60K60), T3 (N60P60K60 + foliar macro–micronutrients), T4 (biofertilizer), and T5 (N60P60K60 + foliar macro–micronutrients + biofertilizer). Integrated foliar fertilization significantly increased nitrogen, phosphorus, and potassium concentrations in vegetative biomass and seeds, leading to higher nutrient uptake and improved nutrient use efficiency compared with mineral fertilization alone. Treatments combining foliar macro- and micronutrients with biofertilizers (T3 and T5) enhanced plant establishment, biomass accumulation, and dry matter allocation to reproductive organs. Seed yield increased from 0.58 to 0.89 t ha−1, while protein and oil contents ranged from 27.0 to 28.4% and 39.8–41.8%, respectively. The combined foliar treatment showed the highest and most stable performance, likely due to improved nutrient uptake and plant growth. These findings indicate that integrated foliar fertilization is an effective and sustainable strategy for improving flax yield stability, nutrient efficiency, and seed quality under semi-arid conditions.

International Journal of Plant Biology doi: 10.3390/ijpb17030018

Authors: Carlos Armas-Díaz David Montesinos-Pereira Lázaro Grisales Maria Corujo José Luis Vázquez-Gutiérrez Daniel Blandón-Granada Eduardo Hernández-Bolaños Andrés Acosta-Pérez Violeta Sánchez-Retuerta Beatriz Porras Laura Cuyas Luis Matías-Hernández

Biostimulants represent a sustainable strategy to enhance the therapeutic potential of medicinal plants, which often exhibit low and variable levels of bioactive compounds. Cannabis sativa, a medicinally important species, produces diverse cannabinoids, such as THC, CBD, CBG, and CBC, whose profiles depend on plant chemotype and determine pharmacological activity. We developed a novel plant-based biostimulant, Tricostimulant™, to optimize cannabinoid production in Cannabis sativa. Field trials demonstrated increased biomass and selective enhancement of cannabinoid content. In high-CBD chemotypes, Tricostimulant™ was associated with higher CBD and CBG without relevant changes in THC levels, whereas in high-THC chemotypes, higher THC values were observed without evident variation in CBD. The most pronounced differences were observed when the biostimulant was applied during the vegetative stage, highlighting the importance of application timing. These results indicate the potential of Tricostimulant™ to modulate cannabinoid profiles, contributing to improved optimization and standardization of cannabis-based therapeutics. Further research is required to confirm these findings and elucidate the underlying mechanisms of biostimulant action.

International Journal of Plant Biology doi: 10.3390/ijpb17030017

Authors: Alexander V. Babosha Pavla O. Loshakova Danila A. Shchelkanov Anastasia D. Alenicheva Margarita M. Gevorkyan Alina A. Pogost

This study used cryoSEM to analyze the seasonal stability of leaf surface micromorphology in cereal hybrids derived from crossing maternal ×Trititrigia cziczinii × Thinopyrum junceum lines with paternal wheat–wheatgrass hybrids. Over two growing seasons, relatively rare traits showed high stability, while most traits exhibiting initial diversity demonstrated seasonal variability. Paternal traits (hairs, prickles, elongated silica cells) predominated in hybrids, and hybrid diversity correlated significantly with paternal, but not maternal, line diversity. In 2025, a significant decrease in some paternally specific traits and an increase in rounded silica cells were observed compared to 2024. Coordinated dynamics were revealed: variations in maternal traits correlated positively with each other and negatively with some paternal traits. While certain micromorphological features exhibited relative stability, employing such traits for taxonomic purposes necessitates caution and a thorough understanding of their inherent variability ranges.

International Journal of Plant Biology doi: 10.3390/ijpb17030016

Authors: Farah Abu Siam Saeid Abu-Romman Saja A. K. Al-Rubaye Ruba M. AL-Mohusaien Monther T. Sadder

Ascorbate peroxidase (APX) is a heme-containing enzyme involved in hydrogen peroxide (H2O2) detoxification within the ascorbate–glutathione (AsA–GSH) cycle. In this study, the full-length genomic DNA and cDNA of an APX1 gene (VsAPX1) were cloned and characterized from Vicia sativa. The genomic sequence of VsAPX1 is 2425 bp in length and comprises 10 exons separated by nine introns, with the first intron located within the 5′ untranslated region (5′UTR). The corresponding cDNA is 1010 bp long and includes a 61 bp 5′UTR, a 753 bp open reading frame, and a 196 bp 3′UTR. VsAPX1 encodes a predicted cytosolic APX protein of 250 amino acids, with a molecular weight of 27.1 kDa and a theoretical isoelectric point (pI) of 5.60. Bioinformatics analysis revealed that the deduced VsAPX1 protein shares high sequence similarity with cytosolic APX1 proteins from other plant species, contains conserved APX domains, and clusters within the cytosolic APX clade in phylogenetic analysis. Quantitative real-time PCR analysis showed that VsAPX1 expression exhibits transient and moderate changes in response to abiotic stress and phytohormone treatments. Transcript levels increased at early time points following heat stress (42 °C), abscisic acid, and salicylic acid treatments, and after 4 h of jasmonic acid exposure, whereas hydrogen peroxide treatment resulted in a gradual down-regulation of expression. Overall, this study provides the first molecular and expression characterization of a cytosolic APX1 gene from Vicia sativa and establishes a foundation for future functional analyses of antioxidant genes in this species.

International Journal of Plant Biology doi: 10.3390/ijpb17030015

Authors: Ali Ait Youssef Reda Meziani Samira Serghat Timothy O. Jobe Mohamed Fokar Allal Douira

Date palms, a vital Moroccan crop that typically flowers once a year, displayed a rare double flowering phenomenon in the summer of 2023. This study investigated the occurrence of this phenomenon across three small oases in the Zagora region of southeastern Morocco. Field surveys revealed that 60% of spring-blooming palms also produced a second bloom in July, affecting trees of all ages, sexes, and varieties. This secondary flowering cycle featured a compressed development period, leading to limited fruit enlargement and a failure of most fruit to reach maturity. Analysis suggests that climatic fluctuations, specifically a delayed temperature rise during the normal spring cycle and relatively cooler shifts in July 2023, likely disrupted the palms’ reproductive schedule, triggering the anomaly. Despite the failed second harvest, the phenomenon did not have a negative impact on the palms’ productivity for the subsequent year, confirming that double flowering is a complex, climate-influenced event that requires further research to understand its full implications for local date production.

International Journal of Plant Biology doi: 10.3390/ijpb17030014

Authors: Nataliia Tkachuk Volodymyr Antonenko Svitlana Kyriienko Erzsébet Kohut Anita Szikura Yurii Karpenko Oleksandr Yakovenko Oksana Sahach Yaroslav Novikov Oleksandr Lukash

The search for new growth-regulating compounds remains a relevant research direction in view of the issue of food security. Previously, a number of thiazolyl acetic acid derivatives have been synthesized, which are promising biologically active compounds according to their physicochemical characteristics. The aim of this work was to study the growth-regulating properties of both previously and newly synthesized thiazolyl acetic acid derivatives using a growth phytotest with Lepidium sativum and Sinapis alba. This study was carried out under laboratory conditions of phytotesting growth indicators of test plants of the class Dicotyledones—L. sativum and S. alba under the influence of 10 μg/mL, 100 μg/mL, and 1000 μg/mL aqueous solutions of the compounds with a comparative analysis of the action of the active components of RhizoponTM and herbicide benazolin. It was found that, exhibiting high herbicidal properties at a concentration of 1000 μg/mL (primarily the compound with a benzothiazole fragment), the studied thiazolyl acetic acid derivatives with decreasing concentration (100 μg/mL and 10 μg/mL) reduce phytotoxicity to its complete elimination against L. sativum and S. alba for the compound 2-[2-(N-tert-butoxycarbonyl)-aminoethyl-1,3thiazol-4-yl]acetic acid (compound 3), as well as against L. sativum for the compound 5-phenylthiazol-2-yl acetic acid (compound 5). A weak stimulating effect on the length of the above-ground part of S. alba seedlings was recorded for 2-[2-(3,3,3-trifluoropropyl)-1,3thiazol-4-yl]acetic acid (compound 1) and 2-[2-(N-tert-butoxycarbonyl)-aminoethyl-1,3thiazol-4-yl]acetic acid (compound 3) at their concentration of 10 μg/mL; however, an improvement in the vitality index under the action of these compounds was not observed. The results obtained in this study indicate the biological activity of both previously and newly synthesized thiazolyl acetic acid derivatives, namely their growth-regulatory properties, expanding knowledge about promising herbicidal compounds with a possible hormetic effect, which requires further research.

International Journal of Plant Biology doi: 10.3390/ijpb17020013

Authors: Gleyse Lopes Fernandes de Souza Francisco Thiago Coelho Bezerra Rejane Maria Nunes Mendonça Walter Esfrain Pereira Marlene Alexandrina Ferreira Bezerra Tayd Dayvison Custódio Peixoto José Adeilson Medeiros do Nascimento Roberto Ítalo Lima da Silva Thayná Kelly Formiga de Medeiros Maria Alaíne da Cunha Lima Daniela Rosario de Mello Anne Alícia Meneses Ferreira Santana Antenor Pereira de Araújo Neto Anna Paula Marques Cardoso Lourival Ferreira Cavalcante

Salinity causes morphophysiological changes that compromise the growth and production of cultivated species, such as Passiflora edulis. However, wild species better tolerate environmental adversities. Therefore, we evaluated the performance of P. edulis scion (Guinezinho, BSR YG1, BRS SC1) grafted on Passiflora rootstocks (P. cincinnata, P. foetida, and P. edulis), regarding the electrical conductivity of irrigation water (0.5 and 3.0 dS m−1), and it was hypothesized that rootstocks would mitigate the effects of salinity and that there is a difference between non-grafted cultivars. Grafted plants reached the trellis stage sooner, and the use of P. foetida rootstocks reduced the time to prune the stem. The highest productivity was obtained with the YG1 cultivar grafted onto P. foetida, under irrigation with water salinity, reaching 8073.96 kg ha−1 and exceeding by up to 139.19% the other grafting and electrical conductivity of the irrigation water. The grafting technique proved effective in mitigating the effects of salinity, particularly the combination between P. foetida and YG1 cultivar P. edulis, which showed compatibility and superior agronomic performance under saline stress conditions. These results indicate a promising strategy for the sustainability of yellow passion fruit cultivation in semi-arid regions, contributing to the maintenance of productivity, even in environments with restricted water quality resources.

International Journal of Plant Biology doi: 10.3390/ijpb17020012

Authors: Jaqueline Zuta Lopez Rosalynn Y. Rivera Elver Coronel Castro Nixon Haro Gerson Meza-Mori Oscar Gamarra Manuel Oliva-Cruz Carlos A. Amasifuen Guerra José Giacomotti Elí Pariente

Globally, coffee-based agroforestry systems are recognized for their capacity to integrate agricultural production with biodiversity conservation, particularly in tropical landscapes under intense anthropogenic pressure. However, significant knowledge gaps remain regarding floristic composition, arboreal structure, and the ecological importance of woody species in Andean agroforestry systems of the Peruvian Amazon, especially along altitudinal gradients. The objective of this study was to characterize the diversity, floristic composition, arboreal structure, and ecological value of woody species in coffee-based agroforestry systems in the Department of Amazonas, Peru. Forest inventories were conducted in twelve one-hectare plots, recording dasometric variables, estimating diversity indices, analyzing floristic affinity, and calculating the Importance Value Index of species. A total of 57 tree species belonging to 41 genera and 25 families were recorded, with moderate diversity levels and a marked dominance of species from the Fabaceae family. The structure showed a predominance of young individuals, concentrated in low and intermediate diameter and height classes, and a moderate shade cover suitable for coffee cultivation. The species with the highest ecological and productive value were Pinus tecunumanii, Colubrina glandulosa, Clitoria juninensis, Inga edulis, and Inga mendozana, which perform key functions related to shade provision and soil fertility. These results are transferable to other coffee agroforestry systems in tropical montane regions and provide relevant evidence for sustainable forest management, biodiversity conservation, and productive optimization, issues of international interest in the agricultural and agroforestry sectors.

International Journal of Plant Biology doi: 10.3390/ijpb17020011

Authors: Maria Gancheva Aleksandr Tkachenko

Oxalis tuberosa (oca) is a tuber crop native to the Andes, valued for its nutrition but understudied genetically. Its strict short-day (SD) tuberization suggests a photoperiodic control mechanism similar to that of potato, where an FT-like protein acts as a mobile “tuberigen” signal. To identify this key regulator, we generated a de novo genome assembly for oca using long- and short-read sequencing. Integrated transcriptomic analysis of leaves under long-day (LD) and SD conditions, along with stems, roots, and tubers, enabled gene annotation and expression analysis. Our study focused on the Phosphatidylethanolamine-Binding Protein (PEBP) gene family, the source of florigen and tuberigen signals. We identified 23 OtPEBP genes and characterized their expression patterns. Among these, we discovered three FT-like homologs that are specifically and strongly upregulated in leaves under SD conditions. We therefore propose these genes as the prime candidates for the mobile tuberigen signal in oca. This work provides the foundational genomic resource for O. tuberosa and advances our understanding of the conserved photoperiodic network controlling storage organ formation beyond the Solanaceae family.

International Journal of Plant Biology doi: 10.3390/ijpb17020010

Authors: Marly Guelac-Santillan Jherson Rojas-Vargas Elmer Chávez-Chacón Eryka Gaslac-Zumaeta Manuel Oliva-Cruz Angel F. Huaman-Pilco

Anthracnose is one of the most destructive diseases of avocado worldwide; however, foliar infections remain poorly documented in Peru. We investigated the etiology of necrotic leaf lesions observed in avocado plantations in Amazonas and assessed the in vitro antifungal activity of two Piper essential oils (EOs). Incidence was quantified in a 420-tree plot (n = 150 plants). Two representative isolates (MGS03, MGS04) were obtained from symptomatic leaves and characterized by morphology and multilocus sequencing (ITS, CHS-1, ACT, and TUB2). Maximum-likelihood phylogeny within the C. gloeosporioides species complex clustered with C. fructicola. Pathogenicity tests on detached leaves (unwounded inoculation) reproduced field symptoms and fulfilled Koch’s postulates. The antifungal activity of Piper carpunya and P. aduncum EOs (50–1000 µL L−1) was evaluated using poisoned-medium assays, revealing a strong dose-dependent inhibition of mycelial growth. Inhibition reached >89.8% at 500 µL L−1 and ~100% at 1000 µL L−1, with P. aduncum showing slightly higher activity. This study provides the first confirmed record of C. fructicola associated with necrotic leaf lesions in avocado in Peru and demonstrates the promising antifungal potential of Piper EOs as eco-friendly candidates for anthracnose management.

International Journal of Plant Biology doi: 10.3390/ijpb17020009

Authors: Sanja Puljas Juraj Kamenjarin Ivica Šamanić

Cynipid gall wasps are known for their ability to manipulate host plant development, redirecting undifferentiated tissues into complex, highly specialised structures. In this study, we investigated how Andricus quercustozae larvae manipulate axillary bud tissues of Quercus virgiliana across four key stages of gall development: initiation, differentiation and growth, maturation, and lignification. Using detailed histological analyses, we characterised progressive tissue differentiation within galls, focusing on the organisation of nutritive, protective, and vascular tissues. Gall development was marked by sustained hyperplasia and hypertrophy, extensive vascular proliferation, and progressive cell wall lignification, resulting in a complex organ optimised for larval nutrition and protection. To complement these anatomical observations, we conducted a preliminary transcriptomic comparison between gall tissue and unmodified leaf tissue. Gene expression analyses revealed suppression of photosynthesis-related functions and coordinated modulation of developmental, regulatory, and metabolic pathways, consistent with a transition from assimilatory leaf tissue to a specialised nutrient sink. Integration of anatomical and transcriptomic evidence supports a model in which cynipid gall wasps intervene at key regulatory nodes of bud development, progressively reprogramming host tissues to form a functionally autonomous gall. These findings provide new insight into the extended phenotype and highlight the plasticity of plant developmental programmes under insect control.

International Journal of Plant Biology doi: 10.3390/ijpb17010008

Authors: Mariya Todorova Zhelyazkova

Lavender has been cultivated in Bulgaria for over a century. The high essential oil content and quality of Bulgarian lavender varieties have established the country as a leading global producer. Studies into the crop’s genetic diversity are essential for selecting varieties best suited to specific environmental conditions, maximizing resilience and yield. Therefore, identifying appropriate genetic markers to monitor lavender diversity is a key prerequisite for developing effective crop selection strategies, particularly in response to the challenges posed by global climate change. In this study, we evaluate the versatility of markers for assessing genetic diversity of lavender genotypes. A total of 96, 97 and 96 bands were recorded using the 13 Start Codon Targeted Polymorphism (SCoT), 13 Inter-Simple Sequence Repeat (ISSR) and 14 Cis-Element Aligned Polymorphism (CEAP) primers, respectively. All amplification programs used were successful in the studied genotypes. Additionally, four informative primers of each marker system were applied for assessment of the within-field genetic variability in two lavender plantations from Bulgaria. This is the first report on the combined use and comparison of CEAP, SCoT and ISSR primers in lavender genotypes in Bulgaria.

International Journal of Plant Biology doi: 10.3390/ijpb17010007

Authors: Unius Arinaitwe Dalitso Noble Yabwalo Abraham Hangamaisho Shillah Kwikiiriza Francis Akitwine

Micronutrients, particularly boron (B), iron (Fe), manganese (Mn), and zinc (Zn), are pivotal for cotton (Gossypium spp.) growth, reproductive success, and fiber quality. However, their critical roles are often overlooked in fertility programs focused primarily on macronutrients. This review synthesizes recent advances in the physiological, molecular, and agronomic understanding of B, Fe, Mn, and Zn in cotton production. The overarching goal is to elucidate their impact on cotton nutrient use efficiency (NUE). Drawing from the peer-reviewed literature, we highlight how these micronutrients regulate essential processes, including photosynthesis, cell wall integrity, hormone signaling, and stress remediation. These processes directly influence root development, boll retention, and fiber quality. As a result, deficiencies in these micronutrients contribute to significant yield gaps even when macronutrients are sufficiently supplied. Key genes, including Boron Transporter 1 (BOR1), Iron-Regulated Transporter 1 (IRT1), Natural Resistance-Associated Macrophage Protein 1 (NRAMP1), Zinc-Regulated Transporter/Iron-Regulated Transporter-like Protein (ZIP), and Gossypium hirsutum Zinc/Iron-regulated transporter-like Protein 3 (GhZIP3), are crucial for mediating micronutrient uptake and homeostasis. These genes can be leveraged in breeding for high-yielding, nutrient-efficient cotton varieties. In addition to molecular hacks, advanced phenotyping technologies, such as unmanned aerial vehicles (UAVs) and single-cell RNA sequencing (scRNA-seq; a technology that measures gene expression at single-cell level, enabling the high-resolution analysis of cellular diversity and the identification of rare cell types), provide novel avenues for identifying nutrient-efficient genotypes and elucidating regulatory networks. Future research directions should include leveraging microRNAs, CRISPR-based gene editing, and precision nutrient management to enhance the use efficiency of B, Fe, Mn, and Zn. These approaches are essential for addressing environmental challenges and closing persistent yield gaps within sustainable cotton production systems.

International Journal of Plant Biology doi: 10.3390/ijpb17010006

Authors: Weverton Gomes da Costa Hélcio Duarte Pereira Gabi Nunes Silva Aluizio Borém Eveline Teixeira Caixeta Antonio Carlos Baião de Oliveira Cosme Damião Cruz Moyses Nascimento

Genome-wide association studies (GWAS) are essential for identifying genomic regions associated with agronomic traits, but Linear Mixed Model (LMM)-based GWAS face challenges in capturing complex gene interactions. This study explores the potential of machine learning (ML) methodologies to enhance marker identification and association modeling in plant breeding. Unlike LMM-based GWAS, ML approaches do not require prior assumptions about marker–phenotype relationships, enabling the detection of epistatic effects and non-linear interactions. The research sought to assess and contrast approaches utilizing ML (Decision Tree—DT; Bagging—BA; Random Forest—RF; Boosting—BO; and Multivariate Adaptive Regression Splines—MARS) and LMM-based GWAS. A simulated F2 population comprising 1000 individuals was analyzed using 4010 SNP markers and ten traits modeled with epistatic interactions. The simulation included quantitative trait loci (QTL) counts varying between 8 and 240, with heritability levels set at 0.5 and 0.8. These characteristics simulate traits of candidate crops that represent a diverse range of agronomic species, including major cereal crops (e.g., maize and wheat) as well as leguminous crops (e.g., soybean), such as yield, with moderate heritability and a high number of QTLs, and plant height, with high heritability and an average number of QTLs, among others. To validate the simulation findings, the methodologies were further applied to a real Coffea arabica population (n = 195) to identify genomic regions associated with yield, a complex polygenic trait. Results demonstrated a fundamental trade-off between sensitivity and precision. Specifically, for the most complex trait evaluated (240 QTLs under epistatic control), Ensemble methods (Bagging and Random Forest) maintained a Detection Power (DP) exceeding 90%, significantly outperforming state-of-the-art GWAS methods (FarmCPU), which dropped to approximately 30%, and traditional Linear Mixed Models, which failed to detect signals (0%). However, this sensitivity resulted in lower precision for ensembles. In contrast, MARS (Degree 1) and BLINK achieved exceptional Specificity (>99%) and Precision (>90%), effectively minimizing false positives. The real data analysis corroborated these trends: while standard GWAS models failed to detect significant associations, the ML framework successfully prioritized consensus genomic regions harboring functional candidates, such as SWEET sugar transporters and NAC transcription factors. In conclusion, ML Ensembles are recommended for broad exploratory screening to recover missing heritability, while MARS and BLINK are the most effective methods for precise candidate gene validation.

International Journal of Plant Biology doi: 10.3390/ijpb17010005

Authors: Aleksandra Yu. Kroupina Pavel Yu. Kroupin Mariya N. Polyakova Malak Alkubesi Alana A. Ulyanova Daniil S. Ulyanov Natalya Yu. Svistunova Alina A. Kocheshkova Gennady I. Karlov Mikhail G. Divashuk

Speed breeding technologies offer a promising avenue for accelerating crop improvement, yet their application to biennial crops like sugar beet remains constrained by extended generation cycles. This study examined the effects of supplemental phosphorus-potassium (PK) nutrition on the development of two hybrids under a speed-breeding protocol. Plants received one of four nutritional regimes: PK supplementation, potassium (K) supplementation, standard Knop’s solution (KS), or nutrient deficiency (D). Digital phenotyping confirmed that adequate nutrition maintained photosynthetic health, as deficiency significantly reduced NDVI and increased PSRI by 75 days. The most notable, genotype-specific effects were observed in reproductive architecture. PK nutrition significantly increased the median number of flower stalks by 17% in Smart Iberia KWS (21.0 vs. 18.0) and substantially in Dubravka KWS (33.0 vs. 1.0). PK also supported root development, increasing mini-steckling weight by 45–183% under white light. In the generative phase, plants under PK nutrition consistently showed the highest progression to flowering and capsule formation. A consistent increase in median 1000-seed weight of 24–36% was associated with PK treatment. In conclusion, supplementing standard nutrition with phosphorus and potassium enhances key yield-related architectural traits and supports reproductive development in sugar beet under speed-breeding conditions, with the magnitude of response depending on genotype. This provides a practical basis for optimizing mineral nutrition to improve the efficiency of accelerated breeding protocols. This provides a practical basis for optimizing mineral nutrition to improve the efficiency of speed breeding protocols.

International Journal of Plant Biology doi: 10.3390/ijpb17010004

Authors: Nikolay Velkov Stanislava Grozeva

Systems favoring cross-pollination, such as male sterility and female flowering type, are of great importance in the development of new hybrid cultivars and their seed production. The advantages of male sterility are expressed in the production of cheaper and competitive seeds. The presence of this characteristic in watermelon is not common, and in some cases, it is accompanied by negative manifestations. A collection of 150 watermelon genotypes was tested at the Maritsa Vegetable Crops Research Institute, Bulgaria, over the past nine years to search for a genetic source of male sterility. The results revealed that two mutations were found. The first mutation was in a plant of the Asar variety, which formed completely degenerated structures in the place of male and female flowers that were completely sterile. The other mutation affected male flowers, female flowers, and leaf shape. Male flowers produced a small amount of pollen. Female flowers were formed, but they were sterile and aborted at an early stage. The genotype can be propagated by pollination of the normal plants, which in the next generation segregate into mutant—25% and normal—75%. The gene source is phenotyped according to the main characteristics of the fruits and the vegetation period. The mutation found cannot be directly used in a breeding program, but it is of interest for studying this important trait. The success of detecting flowers that are sterile depends on the number of watermelon plants, which, for the conditions of the experiment, amounted to a minimum of 4492 plants at a probability level of P3—0.95.

International Journal of Plant Biology doi: 10.3390/ijpb17010003

Authors: Gilberto Garcia Fernando Montero Maria Elena Torres Selwyn Alvarez Wildo Vasquez Abraham Villantoy Yoel Ruiz Fernando Escobal Hector Cántaro-Segura Omar Paitamala Daniel Matsusaka

Purple corn (Zea mays L.) is a nutraceutical crop of increasing economic importance in Peru, yet its productivity is highly influenced by genotype × environment (G × E) interactions across heterogeneous agro-ecological zones. Therefore, selecting suitable genotypes for specific environments is essential to optimize variety deployment and maximize site-specific yield. Five purple-maize genotypes (INIA-601, INIA-615, Canteño, PMV-581, and Sintético-MM) were evaluated in four contrasting Peruvian sites using a randomized complete-block design. Grain yield, field weight, anthesis–silking interval (ASI), plant height, and ear-rot incidence were analyzed with combined analysis of variance (ANOVA), the additive main effects and multiplicative interaction (AMMI), genotype and genotype-by-environment (GGE) biplots, Weighted Average of Absolute Scores (WAAS), weighted average of absolute scores and best yield index (WAASBY), and Y × WAAS indices. Environment accounted for 90.1% of field-weight variation (p < 0.0001) and 50.2% of grain-yield variation (p < 0.001), while significant G × E interactions (3.93% and 18.14%, respectively) justified bilinear modeling. AMMI1 and GGE “which-won-where” biplots identified INIA-615 and PMV-581 as broadly adapted, with INIA-615 achieving the highest WAASBY and positioning in quadrant IV of Y × WAAS (high yield, high stability). INIA-601 and Sintético-MM exhibited exceptional stability (low ASV) but moderate productivity; Canteño showed limited adaptability. Chumbibamba emerged as a key discriminating, high-productivity location. From an agronomic perspective, INIA-615 is recommended for high-productivity valleys such as Sulluscocha and Santa Rita, where its yield potential and stability are maximized. These findings underscore the potential of integrating multivariate stability metrics with physiological and disease-resistance traits to guide the selection of superior purple corn cultivars. Overall, INIA-615 represents a robust candidate for enhancing yield stability, supporting sustainable intensification, and expanding the nutraceutical value chain of purple corn in the Andean highlands.

International Journal of Plant Biology doi: 10.3390/ijpb17010002

Authors: Zhuldyz Batykova Aida Kistaubayeva Malika Abdulzhanova Gulina Doktyrbay Laila Saidullayeva Zhamila Baimirzayeva Moldir Turaliyeva Zhuldyz Ibraimova

The increasing environmental challenges facing modern agriculture necessitate development of sustainable, eco-friendly alternatives to chemical inputs. This study aimed to isolate and characterize rhizophilic bacterial strains from the rhizosphere of the maize hybrid Turan 480 SV (Zea mays L.), with a focus on their plant growth-promoting and biocontrol traits. A total of 23 bacterial isolates were obtained, including 15 Gram-negative and 8 Gram-positive strains. Among these, three strains—CR14, CR18 and CR22—were selected for detailed analysis. All three demonstrated significant indole-3-acetic acid (IAA) production, phosphate and zinc solubilization, nitrogen fixation and antifungal activity. CR14 synthesized 56.01 mg L−1 of IAA and demonstrated the highest zinc solubilization, while CR18 exhibited superior phosphate solubilization and protease activity. CR22 produced the highest IAA (61.46 mg L−1) and demonstrated strong cellulase and amylase activity. In antagonism tests, CR14 suppressed Alternaria alternata with an 80 mm inhibition zone, while CR18 and CR22 effectively inhibited both A. alternata and Fusarium graminearum. Phylogenetic analysis based on 16S rRNA sequencing identified CR18 as Serratia quinivorans, CR14 as Pantoea agglomerans and CR22 as Pantoea sp. The functional diversity of rhizobacteria holds promise as bioinoculants for enhancing maize growth and protecting against soil-borne pathogens in sustainable agriculture.

International Journal of Plant Biology doi: 10.3390/ijpb17010001

Authors: Isaac Theophile Ndjepel Yetnason Adrian Christopher Brennan Dorothy Tchatchoua Tchapda Chimene Abib Fanta

(1) Background: In the Sudano-Sahelian zone of Cameroon, which is affected by drought and forest decline, Vachellia nilotica leaves and seeds are fodder for livestock. (2) Methods: A provenance and progeny study on growth performance and heritability of V. nilotica was carried out to provide a reliable database for tree selection, improvement programs, and the creation of future forested areas in this region. Open-pollinated seeds from 120 mother trees (10 half-sib families per provenance) representing twelve provenances, 50–100 km apart, were used for a progeny trial near Maroua, the Far North region of Cameroon. The experimental design was a Fisher block. (3) Results: The results reveal significant differences among provenances only for the number of leaves, and the variability was marked by coefficients of variation ranging from 0.24−0.63. Narrow-sense heritability was measured, varying from 0.01 ± 0.009 to 0.74 ± 0.02, and genetic gain reached 21.83 at the selection intensity of 5% for the number of leaves per plant. The phenotypic coefficient of variation varied between 14% and 90%. Half-sib families were classified into three subgroups using hierarchical ascending classification, and provenances were grouped into five groups using principal component analysis. (4) Conclusions: These results could contribute to initiating tree selection, but more provenances, longer-term experiments, and molecular genetic testing are needed to complement these nursery-level observations.

International Journal of Plant Biology doi: 10.3390/ijpb16040142

Authors: Angel F. Huaman-Pilco Nicola Fiore Oscar P. Hurtado-Gonzales Larissa Carvalho Costa Xiaojun Hu Manuel Oliva-Cruz Jorge R. Díaz-Valderrama Alan Zamorano

Preserving Peruvian cacao germplasm requires preventing the spread of pathogens such as viruses, yet cacao viral diseases in Peru remain poorly studied. In this study, we characterized the viral sequences associated with native cacao trees from the department of Amazonas, northwestern Peru. Leaf samples from two symptomatic plants (mosaic, yellowing, leaf deformation) and one asymptomatic plant were collected from the cacao germplasm bank of the Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas. RNA high-throughput sequencing identified four RNA segments consistent with the genus Emaravirus: RNA1 (7142 nt; replicase P1), RNA2 (2225 nt; glycoprotein P2), RNA3 (1269 nt; nucleocapsid P3), and RNA4 (1286 nt; movement protein P4), sharing 32.6–45.9% amino acid identity with European mountain ash ringspot-associated emaravirus (EMARaV). Phylogenetic analysis of P1–P4 proteins placed this virus in a distinct lineage, confirming it as a novel species, Theobroma cacao emaravirus A (ThCEV-A). Specific RT-PCR detected ThCEV-A in 11 additional accessions, with symptoms including yellow mosaic and mottling. This study documents for the first time the presence of a novel Emaravirus in cacao, highlighting the need to assess its epidemiology, vector(s), and potential impact on cacao production in its center of origin.

International Journal of Plant Biology doi: 10.3390/ijpb16040141

Authors: Rongrong Tan Long Jiao Peizhuo Wu Danjuan Huang Hongjuan Wang Xun Chen Yingxin Mao

The tea plant (Camellia sinensis (L.) O. Kuntze) is a globally important crop, yet its cultivation is continually challenged by a range of viral pathogens that can compromise plant health and product quality. In this study, eighteen symptomatic leaves were collected from the Hubei Province Tea Germplasm Resources Nursery, China, representing multiple cultivars and diverse genetic backgrounds. The samples were pooled into three groups and subjected to ribodepleted transcriptome sequencing. Analyses revealed a complex virome, with Tea plant necrotic ring blotch virus (TPNRBV) dominating Pools A and B, whereas Badnavirus betacolocalasiae was the most prevalent in Pool C. Functional enrichment of viral genes indicated involvement in multiple biological processes, including replication, host interaction, and metabolism. Notably, two previously uncharacterized viruses were identified: Tea plant-associated ourmia-like virus 1 (TeaOLV1) and Tea plant-associated rhabdo-like virus 1 (TeaRLV1). Phylogenetic reconstruction positioned TeaOLV1 within the Penoulivirus genus, while TeaRLV1 formed a distinct clade among plant-associated rhabdoviruses. Conserved motif analysis revealed typical viral domains, accompanied by lineage-specific variations in tea plants. Collectively, these findings enhance our understanding of the viral diversity in tea plants, provide refined taxonomic placement for newly identified viruses, and offer molecular insights into their evolutionary relationships and potential functional roles.

International Journal of Plant Biology doi: 10.3390/ijpb16040140

Authors: Ryan Koeth Shahzad Hussain Shah Calvin Juel Rigney Changbin Chen

Zingeria biebersteiniana, a grass species with the lowest known chromosome number among angiosperms (2n = 2x = 4), offers a distinctive platform for cytogenetic and grass research. Despite its unique karyotype and potential for molecular and educational applications, no transformation system has previously been reported for this species. Here, we establish a reproducible Agrobacterium tumefaciens-mediated transformation protocol for Z. biebersteiniana, optimized through comparative evaluation of three tissue culture media. A modified Khromov medium with Plant Preservative Mixture supported robust callus induction and plant regeneration, enabling the successful introduction of a GFP–mouse talin1 fusion construct driven by the rice Actin-1 promoter. Transgenic lines were validated via PCR amplification of the hygromycin resistance gene, and GFP signals were observed in transformed individuals. However, the expression pattern was less specific than previously reported in rice, potentially due to species-specific differences in mouse Talin1 protein localization. Although actin filament visualization in mature pollen remained unspecific, the protocol provides a foundational tool for future molecular and functional genomics and genetics studies. This work represents the first documented genetic transformation of Z. biebersteiniana, expanding its utility as a model system in plant biology and genomics.

International Journal of Plant Biology doi: 10.3390/ijpb16040139

Authors: Jamille Cardeal da Silva Jailton de Jesus Silva Raquel Araujo Gomes Claudinéia Regina Pelancani Cruz Barbara França Dantas

Commiphora leptophloeos, a native Caatinga species with economic and medicinal potential, faces propagation challenges due to seed dormancy and extractive use. The germination test, the official method for seed quality assessment, is time-consuming, whereas the tetrazolium test (TZT) offers a rapid alternative for determining seed viability. This study aimed to establish and validate a TZT protocol for C. leptophloeos seeds. Seeds collected in 2025 were extracted after natural fruit drying and then stored in a cold chamber. The germination test was conducted with seeds without pyrenes at 30 °C. For the TZT, a completely randomized design was used in a 6 × 4 factorial scheme (six TZT concentrations × four immersion times), with adjustments in seed preparation and staining procedures. Higher concentrations (0.5% and 0.75%) combined with shorter immersion periods (2 h) provided the best results, especially 0.75% for 2 h, which yielded 89% viability. Very low concentrations combined with short periods resulted in little or no staining. Compared with the germination test (35%), the TZT showed greater sensitivity in detecting viable seeds. We conclude that the TZT is highly efficient for assessing the viability of C. leptophloeos seeds, with optimal responses at 0.5–0.75% TTC and 2–4 h immersion periods, and represents a strategic tool to support the conservation and sustainable use of this species.