Plants doi: 10.3390/plants15223540

Authors: Yan Guo Feng Huang

Phyllosphere microbial communities are critical for plant health and ecosystem functioning, yet the relative importance of host plant species and nitrogen deposition in shaping these communities remains unclear. In this study, a two-year greenhouse experiment was conducted with three native subtropical plant species representing distinct evolutionary lineages—Blechnum orientale (Bo, fern), Cryptocarya concinna (Cc, magnoliid), and Calamus simplicifolius (Cs, monocot)—subjected to four levels of anthropogenic nitrogen application (0, 35, 70, and 105 kg N ha−1 yr−1). The leaf physicochemical properties and the richness, diversity, community composition, and structure of phyllosphere fungal and bacterial communities were examined. Leaf total organic carbon (TOC), total nitrogen (TN), potassium (K), sodium (Na), and magnesium (Mg) contents differed significantly among the plant species (p < 0.05), and Bo exhibited the highest fungal and bacterial richness and diversity (e.g., fungal richness Bo: 1292, Cc: 661, Cs: 1224; bacterial richness Bo: 174, Cc: 82, Cs: 59). Principal coordinate analysis (PCoA) further revealed clear separation of fungal and bacterial communities among the three species, with plant species explaining 36.7% and 37.9% of the total variance, respectively (p < 0.01). In contrast, nitrogen application exerted only sporadic effects: it increased leaf TN content only in Cc, increased leaf total phosphorus (TP) content under low nitrogen in Bo, and decreased certain metal elements (Mg in Bo, K in Cs). Notably, bacterial richness in Bo significantly dropped from 174 to 77 under high nitrogen (p < 0.05), while significant shifts in microbial community composition were detected only in the fungal communities of Cs (p < 0.05). Across plant species, and most strongly in Cc under nitrogen application, leaf TP was positively correlated with bacterial richness, diversity, and Gammaproteobacteria (p < 0.001), but negatively with Alphaproteobacteria (p < 0.05). Taken together, these results demonstrate that host plant species is the primary driver shaping phyllosphere microbial communities, whereas short-term nitrogen addition exerts only sporadic and species-dependent (only for leaf bacterial communities) effects, reflecting the intrinsic resistance of phyllosphere microbial communities to nitrogen deposition over short time scales.

Plants doi: 10.3390/plants15223539

Authors: Aslıhan Cilingir Tutuncu Burhan Ozturk Esin Hazneci Koray Korkmaz Harun Ozer Coşkun Gülser

This two-year study evaluated the effects of rainbow trout (Oncorhynchus mykiss) waste-derived protein hydrolysates (HYDs), applied to the roots or foliage of soilless-grown tomato plants at 0.2 (HYD1), 0.4 (HYD2) and 0.8 g L−1 (HYD3), on fruit quality during 28 days of cold storage under modified-atmosphere packaging. Because year × treatment interactions were significant, each year was analyzed separately. In both years, HYD treatments reduced weight loss and respiration rate and maintained firmness compared with the control. Root-applied HYD1 was the only treatment that maintained higher firmness than the control from day 7 to day 28 in both years, and it had the highest overall quality index in a multi-criteria (TOPSIS) evaluation at the end of storage in both years. In contrast, the effects on soluble solids, titratable acidity, vitamin C, lycopene, anthocyanins, phenolics, flavonoids and antioxidant activity depended on the year, and vitamin C and lycopene responded in opposite directions in the two years. Fish-waste HYDs, particularly root-applied HYD1, are therefore promising for maintaining firmness and reducing postharvest losses of tomato, whereas their effects on bioactive compounds require further multi-season evaluation.

Plants doi: 10.3390/plants15223538

Authors: Jiayi Chen Qian Zhou Jingbo Zhang Pingping An Xiaoneng Wang Keying Wu Fenglan Liu Haixia Jiang Jing Guo Huixin Zhao

Late-spring cold stress disrupts vegetative physiology in apple (Malus domestica), a major horticultural crop in continental regions such as Xinjiang, China. Current cold-protection strategies, including physical heating and chemical sprays, are costly, labor-intensive, and environmentally unfriendly, while microbial metabolite-based elicitors have emerged as sustainable alternatives but remain underexplored in apple cold tolerance. This study aimed to develop and characterize a novel Bacillus subtilis BS-Z15 metabolite extract (B-MES)-based elicitor (E6) for priming cold tolerance in apple leaves, and to elucidate its molecular mechanisms through integrated transcriptome and metabolome analysis. E6 was optimized using an L18(37) orthogonal array design (18 experimental runs, 7 factors at 3 levels each) and validated across three biological scales (detached branches, potted seedlings, adult-tree-derived branches) in two apple cultivars. E6 pretreatment significantly increased SOD and POD activities and proline content while reducing MDA content and relative electrolyte leakage and preserving chlorophyll under 0 °C stress. Integrated transcriptome and metabolome analysis revealed concurrent activation of PTI–like signaling, phenylpropanoid metabolism, glutathione metabolism, and MAPK signaling pathways. A rosmarinic acid-laccase module showed sustained positive correlation during priming and recovery, which is indicative of a sustained priming response. Cross-cultivar validation in ‘Wanglin No. 1’ confirmed the generality of the protective mechanism. This study provides a novel candidate biostimulant and mechanistic insights into elicitor-induced cold priming in apple leaves; further validation at the reproductive tissue and orchard scale is warranted.

Plants doi: 10.3390/plants15223537

Authors: Ramandeep Kaur Hansaraj Pradhan Raman Dhariwal Vikrant Tyagi Vikas Kumar Ravat Vinod Kumar Mishra Arun Kumar Joshi Neeraj Kumar Vasistha

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a major foliar disease of bread wheat (Triticum aestivum L.). Genomic prediction (GP) can accelerate breeding for this trait, but conventional, machine-learning (ML), and deep-learning (DL) methods have rarely been compared side by side. Using the Wheat Association Mapping Initiative (WAMI) panel (n = 286), phenotyped for powdery mildew severity over three years (DS1–DS3) and genotyped with the Illumina 90K SNP array, we compared 30 GP models under per-year and genotype-grouped pooled cross-validation. Broad-sense heritability ranged from 0.65 to 0.89. In the pooled, leakage-safe analysis, six Bayesian-alphabet models clustered at the top (r = 0.291–0.299), too close to identify a single best model; partial least squares (PLS) regression was the strongest non-Bayesian model, and elastic net was the strongest ML model within each year. Within-year accuracy under random 5-fold cross-validation was higher (up to r ~ 0.70 in DS2) but may be optimistic, as folds were not stratified by genomic relatedness. Accuracy showed limited, non-monotonic sensitivity to marker density and increased, though not strictly monotonically, with training population size. Leave-one-year-out (LOYO) accuracy was near zero (mean r = 0.009); fitting year as a fixed effect did not improve it, and cross-year genetic correlations were low (0.08–0.15), indicating limited shared genetic control across years. DL results apply only to the architectures and sample size tested. With pooled accuracy near r = 0.30, these models suit within-season prediction in this panel, whereas reliable cross-season application is not supported by the present results and will require broader multi-environment training and/or season-specific recalibration.

Plants doi: 10.3390/plants15223536

Authors: Olivera Stajković-Srbinović Mila Pešić Biljana Sikirić Sonja Tošić Jojević Marina Jovković Nikola Koković Zoran Dinić Snežana Andjelković Uroš Buzurović Vesna Mrvić

Nickel (Ni) is an essential micronutrient for plants. It is of particular importance in legumes, but it can be toxic in high concentrations. In this research, the uptake and translocation of Ni and other potentially toxic elements (PTEs) were analyzed in alfalfa (Medicago sativa L.) inoculated with thirteen rhizobial strains (Ensifer spp.) grown in two soils that are naturally high in Ni (58 mg/kg and 195 mg/kg and other PTEs (Cr and Pb) over two years. Four cuttings were used in the pot experiment. The values of all tested elements (As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn) in alfalfa shoots mainly fit within the common concentrations for plants. Significantly higher concentrations of all elements were retained in the alfalfa roots, and the translocation factor (TF) was below 1, with the exception of Cr and Mn (for which the TFs were 1.56 and 1.14, respectively). The Ni concentrations in alfalfa shoot and root varied depending on the strain, soil and cut and ranged from 0.86 to 7.64 mg/kg in shoot dry weight, and from 3.57 to 62 mg/kg in root dry weight, while the TF values for Ni were 0.08–0.81. In the early phase of growth (the first cut in the first year) in both soils, all tested strains reduced the Ni concentrations in alfalfa shoot. Overall, for the whole experimental period, inoculation with strains 4193cs, G-nov, 224, 218, L4 and 4148ss decreased the Ni concentrations in shoots compared to uninoculated plants. The results indicate that rhizobia have the potential to improve alfalfa growth with preferential Ni accumulation in alfalfa roots and, potentially, low translocation in shoots in soils with elevated Ni and other PTE concentrations.

Plants doi: 10.3390/plants15223535

Authors: Ardas Kavaliauskas Lutz Weihermüller Renaldas Žydelis

This two-year study (2023–2024) evaluated maize (Zea mays L.) responses to mineral, organic, and integrated fertilization with hydrogel and silicon at two contrasting sites in Central Lithuania. Thirteen treatments were tested to assess grain yield, total aboveground biomass (TAB), N uptake, soil mineral N dynamics, and agronomic N use efficiency (AEN). AN2 (170 kg N ha−1) produced two-year mean grain yields of 7.33 and 5.73 t ha−1 at Akademija and Ruoščiai, respectively. AN1 + PCM (90 kg mineral N + 80 kg manure N ha−1) yielded 6.55 and 5.14 t ha−1. Both treatments received 170 kg N ha−1, and their grain yields did not differ significantly in the within site-year comparisons (Duncan’s test, p > 0.05), although their P and K inputs differed. The two-year mean AEN values for AN2 and AN1 + PCM were 12.41 and 7.86 kg grain kg−1 N at Akademija, and 9.71 and 6.27 at Ruoščiai, respectively. Organic-only fertilization resulted in lower mean grain yields than AN2, but the year-to-year responses varied, so the cumulative effects cannot be inferred. Across both seasons, TAB and grain yield were lower at Ruoščiai than Akademija by 23.1–26.0 and 18.9–25.8%, respectively; these site differences cannot be attributed to soil texture alone. More residual mineral N remained at Ruoščiai in 2023, but this does not directly measure crop N recovery. Hydrogel and silicon provided minor treatment- and year-dependent numerical yield increases, with no consistent advantage.

Plants doi: 10.3390/plants15223533

Authors: Jun Hu Xing-Yue Zhang Qing Liu Ya Huang Hong-Jing Zhang Si-Rong Yi

Euonymus zhongianus (Celastraceae), a new evergreen shrub from eastern Chongqing, China, is described and illustrated herein. Morphological comparisons provide the primary evidence for recognizing this taxon as an undescribed species. Molecular phylogenetic analyses based on nuclear ribosomal internal transcribed spacer (nrITS) and three plastid markers (matK, psbA-trnH, and trnL-F) clarify its phylogenetic affinity and placement among the sampled taxa. The new taxon is morphologically allied to E. duanensis, E. euscaphis, and E. viburnoides, but can be readily distinguished by its shorter stature, short petioles, thinly leathery leaf blades with fewer lateral veins, elliptically lobed capsules, and an orange-yellow aril enveloping more than two-thirds of the seed. Phylogenetically, E. zhongianus forms a well-supported monophyletic group with E. euscaphis. This species is currently known only from a single locality in Shizhu Tujia Autonomous County. Following the IUCN Red List Categories and Criteria (Version 15.1), it is assessed as Data Deficient (DD).

Plants doi: 10.3390/plants15223534

Authors: Konstantina V. Kocheva Kamen P. Kamenov Elisaveta B. Kirova Miroslav I. Karabaliev Stefka G. Taneva

Electrical impedance spectroscopy (EIS) is a non-destructive analytical technique that characterizes the electrical properties of materials across a broad frequency range. By simultaneously measuring the magnitude and phase of the impedance, EIS enables the determination of resistive, capacitive, and diffusive components, which correspond to intrinsic physical, chemical, and biological processes. EIS has gained increasing attention as a promising tool for soil monitoring, assessment of plant physiological status, and quality evaluation of fruits and vegetables. Plants are often exposed to hostile environments, which have negative impact on their growth and development. As a result, plants have evolved defense mechanisms, which help maintain their metabolism and preserve homeostasis. This review outlines EIS current applications in the investigation of plant responses to abiotic stress. The basic principles of EIS, the processes underlying impedance analysis, data interpretation, and modeling strategies are discussed, while addressing challenges such as environmental variability and heterogeneity of biological samples. By presenting a comprehensive overview, this article highlights the potential of EIS as a tool for monitoring stress-associated physiological changes thereby supporting the development of enhanced sustainable agricultural practices. Finally, future perspectives are considered, including the integration of EIS with smart agriculture technologies, Internet of Things (IoT) devices, and machine learning methods to enable real-time decision support.

Plants doi: 10.3390/plants15223532

Authors: Yibo Zheng Chunhui Mao Huiling Ma

To investigate the physiological and transcriptomic effects of exogenous melatonin (MT) in alleviating alkaline stress in alfalfa, seedlings of Medicago sativa L. cv. ‘Gannong No. 8′ were exposed to 150 mmol/L NaHCO3 and sprayed with 150 μmol/L MT. Growth, antioxidant capacity, membrane damage, photosynthetic pigments, and osmotic adjustment were measured, and transcriptome sequencing was performed to identify differentially expressed genes and pathways. Alkaline stress reduced growth and leaf relative water content (RWC), degraded chlorophyll, and altered antioxidant, membrane-damage, and osmolyte parameters. Under alkaline stress, MT increased catalase (CAT) activity at 24 h, reduced malondialdehyde (MDA) accumulation, maintained total chlorophyll, and improved root fresh weight and RWC measured at 10 d, and reduced leaf Na+ accumulation and Na+/K+ ratios in leaves and roots, although root Na+ remained higher than in A; REC was not reduced. At 12 h, the A-M versus A difference in superoxide dismutase (SOD) activity was not significant. At 24 h, CAT activity in the stress + MT group (A-M) was 5812.5 U g−1 FW, 2.94 times that in A (1980.0 U g−1 FW); MDA was 30.1 nmol g−1 FW and total chlorophyll was 3.03 mg g−1 FW. MT alone reduced shoot fresh weight at 10 d and increased MDA at 12 and 24 h in non-stressed plants, indicating that its effects were context-dependent. Transcriptome analysis identified transcriptional changes in candidate pathways related to glutathione metabolism, MAPK and JA/ABA signaling, phenylpropanoid and isoflavonoid metabolism, and osmotic adjustment, including MsGSTF9, MsJAZ6, MsLOX2.1, MsP5CS, MsHCT, and MsCYP81E1. Weighted gene co-expression network analysis (WGCNA) indicated that modules containing MsGSTF9 and MsJAZ6 were associated with osmotic and antioxidant traits. Overall, MT was associated with improved growth, water status, chlorophyll retention, and higher CAT activity under alkaline stress, but it did not restore membrane permeability, and the transcriptomic pathways remain candidate mechanisms rather than functionally validated processes.

Plants doi: 10.3390/plants15223531

Authors: Yi Song Nan Sun Yanan Chen Xiaoyan Liu Dong Li Bei Li Qingqing Gao Hongxia Zhang Yanfeng Liu Limin Wang

Salt stress severely limits rapeseed growth and productivity, with varietal differences in tolerance, yet the molecular responses remain poorly understood. In this study, we compared the physiological and transcriptomic responses of a salt-tolerant cultivar ‘ZY821’, and a salt-sensitive cultivar ‘HS3’ under salt stress. Physiological analysis confirmed that ‘ZY821’ exhibited significantly higher plant height, shoot and root biomass, chlorophyll content, and SOD and CAT activities, while accumulating lower level of H2O2 under salt stress compared to ‘HS3’. Transcriptome analysis revealed 1542 and 2348 differentially expressed genes (DEGs) in ‘ZY821’ and ‘HS3’ under salt stress, respectively. A formal genotype × treatment interaction analysis further identified 1011 genes with genotype-dependent salt responses. GO and KEGG enrichment analyses indicated that upregulated genes in ‘ZY821’ were primarily associated with environmental sensing, starch and sucrose metabolism, and phenylpropanoid biosynthesis, whereas downregulated genes involved lipid metabolism. In contrast, ‘HS3’ exhibited broad upregulation of ribosome-related genes and downregulation of secondary metabolism and glutathione-mediated redox pathways. Analysis of transcription factors (TFs) identified 569 differentially expressed TFs under salt stress. Basic characterization revealed that three candidate TFs (BnaERF106, BnaNFYA6, and BnaCOL13) localized to the nucleus, with BnaERF106 exhibiting transcriptional activation activity. BnaERF106 and BnaNFYA6 were downregulated in both cultivars under salt stress, while BnaCOL13 showed contrasting gene expression patterns. Heterologous expression of BnaERF106 and BnaCOL13 reduced yeast salt tolerance, consistent with a possible negative regulatory role that requires validation in plants. Collectively, these findings provide a candidate gene resource and testable hypotheses for future functional validation.

Plants doi: 10.3390/plants15223528

Authors: Feifei Zhang Habib Ullah Shah Faisal Iqbal Hussain Xin Pan

Plants must tolerate fluctuating and increasingly concurrent abiotic and biotic stresses while maintaining growth and reproduction. Understanding how plants integrate these environmental cues is therefore central to developing climate-resilient crops. Plant stress tolerance is not governed by isolated pathways but by a multilayered signaling network in which stress perception is translated into calcium (Ca2+), reactive oxygen species (ROS) and reactive nitrogen species (RNS) signals, integrated through phytohormone networks and mitogen-activated protein kinase (MAPK) cascades, and decoded by transcription factors, post-translational modifications and chromatin-based mechanisms. In this Review, we synthesize recent advances from stress perception and early signaling through hormone crosstalk, transcriptional and post-translational regulation, epigenetic control and stress memory, with particular emphasis on the integration of abiotic and biotic stress responses. We highlight how shared signaling components generate both convergence and stimulus-specific outputs, and why responses to combined stresses cannot be reliably predicted from single-stress experiments. We further examine how mechanistic knowledge is being translated into crop improvement through genome editing, synthetic biology, microbiome-based strategies, priming and data-driven breeding. Finally, we identify critical gaps in signal specificity, network integration, stress memory, genotype-by-environment interactions and field validation. We propose that future crop improvement should prioritize conditional, tissue-specific and quantitatively tuned interventions evaluated under realistic multifactorial stress environments. This integrative framework links molecular stress perception and signaling with memory and crop engineering, providing a mechanistic basis for developing crops that maintain productivity under increasingly complex environmental conditions.

Plants doi: 10.3390/plants15223530

Authors: Jia Miao Min Tang Ling Li Hai Tian Wan-Yuan Shi Jing-Jie Huang Ying-Feng Niu Xiao-Long Sun Ming-Chun Gui

Somatic embryogenesis is important for the clonal propagation of elite rubber tree (Hevea brasiliensis) germplasm, yet metabolic changes during embryo development, particularly in triploid cultivars, remain poorly understood. Here, LC–MS-based untargeted metabolomics was used to compare the diploid cultivar Yunyan73477 and the triploid cultivar Yunyan7346 across four stages: mature callus, heart-shaped somatic embryos, cotyledonary somatic embryos, and mature somatic embryos, with three biological replicates per cultivar-stage group. Differential metabolite accumulation analysis, stage-ordered abundance trend analysis, and KEGG pathway enrichment analysis were performed. The cultivars differed in the numbers and patterns of differentially accumulated metabolites (DAMs) during early transitions. During the cotyledonary-to-mature somatic embryo transition, 90 and 64 DAMs were identified in Yunyan73477 and Yunyan7346, respectively; 86 and 60 decreased in relative abundance, including 44 shared DAMs. In both cultivars, metabolites with overall decreasing trends were significantly enriched for isoflavonoid biosynthesis, whereas those with increasing trends were significantly enriched for linoleic acid and alpha-linolenic acid metabolism. Under the applied statistical thresholds, diterpenoid biosynthesis was significantly enriched only among metabolites assigned to the overall increasing STEM Profile 9 in Yunyan7346, including annotated gibberellin-related compounds, although this statistical enrichment does not by itself indicate enhanced diterpenoid pathway activity. These exploratory findings reveal shared stage-associated metabolic patterns and cultivar-associated differences between the two tested cultivars, Yunyan73477 and Yunyan7346, under the applied culture conditions, and provide candidate metabolites for targeted validation and subsequent functional investigation.

Plants doi: 10.3390/plants15223529

Authors: Elma Vuko Ivana Bočina Juraj Kamenjarin Sanja Radman Gabriela Radnić Petra Žilić Željana Fredotović

Pentanema spiraeifolium (formerly Inula spiraeifolia L.) is an aromatic Mediterranean Asteraceae species for which anatomical and chemical information remains limited. This study addresses these gaps by providing new data on leaf anatomy, micromorphology, ultrastructure, histochemistry, and volatile composition, together with a preliminary assessment of hydrolate activity against tobacco mosaic virus (TMV). Leaf structure was examined using light, scanning, and transmission electron microscopy, complemented by histochemical analyses. Volatile compounds in fresh and dried plant material, essential oils, and hydrolates were characterised by gas chromatography–mass spectrometry, and hydrolate activity against tobacco mosaic virus was evaluated using a local lesion assay on Datura stramonium. The leaves showed a dorsiventral organisation, abundant glandular and non-glandular trichomes, and cavity-like structures within the mesophyll showing positive NADI reactions. Dense marginal indumentum and distinctive finger-like epidermal wall projections were also observed. Histochemical analyses indicated lipophilic material in glandular trichomes and the mesophyll, with NADI reactions consistent with the presence of terpenoid compounds, while ultrastructural observations revealed abundant cytoplasmic lipid droplets. GC–MS revealed a chemically diverse, terpene-dominated volatile profile that differed markedly among plant material, essential oils, and hydrolates. In a preliminary TMV assay, hydrolate-treated plants showed 23% fewer local lesions than the water-treated, TMV-inoculated control group, although the difference was not statistically significant. Together, these findings provide new anatomical, micromorphological, ultrastructural, histochemical, and chemical data for P. spiraeifolium, alongside a preliminary biological observation that warrants further investigation.

Plants doi: 10.3390/plants15223525

Authors: Chenfeng Yang Zengcuo Nao Yanmei Wang Shuaishuai Huang Yonghong Zhou Ruojin Liu

Secondary metabolism contributes substantially to the medicinal value and environmental adaptation of plants, yet how natural microhabitat variation is associated with secondary metabolite accumulation remains poorly understood in high-altitude medicinal plants. Here, seven natural populations of Berberis hemsleyana Ahrendt from the northern and southern mountains of Lhasa, spanning an altitude of 3650–3850 m a.s.l., were investigated using soil physicochemical measurements, untargeted metabolomics, transcriptomics, and cross-omics association analyses. All fruits were collected at full maturity to minimize variation associated with developmental stage. At each sampling site, three fruits were pooled into a single sample for transcriptomic analysis, resulting in one RNA-seq library per site and seven libraries in total (n = 1 per site). Therefore, transcriptomic results were interpreted as descriptive site-associated expression patterns rather than statistically validated differential expression. Soil properties and fruit metabolic profiles exhibited pronounced spatial heterogeneity among sampling sites, although 2558 metabolites were shared across all sites, indicating a relatively stable core metabolome with site-associated differences mainly reflected in metabolite abundance variation. Candidate metabolites, including isoquinoline alkaloids and flavonoid/phenylpropanoid-related compounds such as berberine, palmatine, apigenin, and kaempferol, showed distinct accumulation patterns among microhabitats. Candidate metabolite features prioritized using VIP scores > 1 were used for exploratory comparisons, while VIP rankings alone were not considered sufficient to establish statistically significant differential abundance. The RDA model explained a limited proportion of metabolite variation (adjusted R2 = 0.033) and was not statistically significant based on the permutation test (p = 0.301). Exploratory cross-omics analyses suggested candidate associations among MYB and NAC transcription factors, metabolism-related structural genes, and representative metabolites. Overall, this study provides exploratory statistical associations between soil properties and fruit metabolic patterns across natural microhabitats. These associations do not establish causal effects or confirmed regulatory mechanisms and require further validation.

Plants doi: 10.3390/plants15223526

Authors: Bingliang Liu Chen Yang Fengfan Sun Qiang Li Weijun Chen Yang Tao Yujie Yang Xinhui Wang

Plant genotypes can shape rhizosphere microbial communities, yet the host genes linking plant developmental regulation with belowground ecological processes remain poorly understood. LAZ1 proteins are conserved DUF300 transmembrane proteins implicated in plant signaling and stress responses, but their roles in crop growth and rhizosphere interactions are largely unknown. Three independent maize ZmLAZ1-5 overexpression lines (L161, L162, and L163) were evaluated by phenotyping and transcriptome sequencing at the three-leaf stage. Rhizosphere soil properties and bacterial and fungal communities were assessed at a later developmental stage following transplantation into soil. ZmLAZ1-5 overexpression consistently promoted root elongation, with L162 showing a 50.5% increase in root length relative to B73. Transcriptome analysis identified 4439, 1917, and 2225 differentially expressed genes, respectively, including 1451 shared genes enriched in hormone signaling, MAPK signaling, and α-linolenic acid metabolism. In pot experiments, overexpression was associated with line-dependent changes in rhizosphere nutrient status, with AN, AP, and AK contents consistently higher in all three transgenic lines. Bacterial and fungal α-diversity changed little, whereas community composition and predicted functional profiles showed genotype-associated shifts. These findings identify ZmLAZ1-5 as a positive regulator of maize root elongation and extend the functional scope of DUF300 proteins to host genotype-associated rhizosphere ecological responses.

Plants doi: 10.3390/plants15223527

Authors: Nik Mahnič Barbara Jeršek Marjeta Mencin Urška Vrhovšek Matej Bernard Kobav Klemen Bohinc Rajko Vidrih

Blue light-emitting diode (LED) light can suppress the postharvest pathogen Penicillium expansum (P. expansum), but its effects on the phenolic response of infected apple skin remain poorly understood. In this study, ‘Idared’ apples were inoculated with P. expansum and incubated under continuous blue LED light irradiation or in darkness. Perilesional skin tissue was sampled at 2 days post-inoculation (dpi), while at 7 dpi, both perilesional tissue and an adjacent outer ring were sampled. Twenty phenolic compounds were quantified by UPLC-MS/MS and UPLC-PDA-QDa. A distinct red halo developed around inoculation sites only under blue LED light. At 2 dpi, the total concentration of quantified phenolics in apple skin was 22.6% higher under blue LED light than in darkness. Cyanidin 3-O-galactoside was 10.3-fold higher. Three minor cyanidin glycosides were detected only in irradiated tissue. At 7 dpi, cyanidin 3-O-galactoside was 5.4-fold higher in irradiated than in dark-incubated perilesional tissue. Chlorogenic acid was strongly concentrated in perilesional tissue relative to adjacent outer tissue, with concentrations 3.7-fold higher under blue LED light and 2.5-fold higher in darkness. In contrast, several flavan-3-ols, procyanidins, phloridzin, and naringenin 7-O-glucoside were more abundant in the adjacent outer tissue. Thus, total phenolic concentrations did not capture these contrasting spatial patterns. The red halo was associated with a localized anthocyanin signature within a broader, compound-specific phenolic response to P. expansum infection under blue LED light.

Plants doi: 10.3390/plants15223524

Authors: Jun Wang Hao Wang Simin Sheng Weiyi Li Pengfei Cheng Wenbiao Shen

Argon has recently been considered a potential positive regulator of alfalfa tolerance against cadmium (Cd) stress, and its response is dependent on nitric oxide (NO) production and signaling. However, whether or how argon-driven NO influences plant metabolism for Cd tolerance is largely unknown. To answer this scientific question, integrated transcriptomics and physiological analyses were adopted. In the presence of Cd, the transcriptomes showed that the differentially expressed genes (DEGs) controlled by argon-enriched solution (ARW + Cd) were classified, indicating that argon influences intensive cellular metabolic reprogramming. After the removal of endogenous NO using the NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), DEG-related photosynthesis, phenylpropanoid biosynthesis, and plant hormone signal transduction in the ARW + PTIO + Cd group were impaired. Biochemical experiments showed that photosynthetic pigments and the representative genes related to phenylpropanoid biosynthesis were stimulated by argon, but were sensitive to PTIO addition. Similar alterations were observed in the transcripts of several important gene-encoding enzymes involved in phenylpropanoid biosynthesis, including shikimate O-hydroxycinnamoyl transferase (HCT), caffeoyl-CoA O-methyltransferase (CCoAOMT), and cinnamoyl-CoA reductase (CCR). Contents of phenolic acids (cinnamic acid, p-coumaric acid, and caffeic acid) and some lignin monomers (5-hydroxy-guaiacyl lignin and syringyl lignin) were accordingly increased by argon-driven NO. In conclusion, integrated transcriptomics and biochemical analyses reveal that argon-driven NO contributes to the regulation of phenylpropanoid metabolism, thus leading to cadmium tolerance. This study extends our previous findings and provides a basis for further understanding of the argon-based biology in plants.

Plants doi: 10.3390/plants15223523

Authors: Ke Hu Jiaxing Song Shuyun Zhao Huajiang Liu Yuxiang Wang Jing Yang

Drought adaptation in desert C4 species requires coordinated physiological and transcriptional regulation, but the temporal progression of these responses remains poorly resolved in Kochia prostrata. In this study, progressive drought stress was imposed by withholding irrigation for 7, 14, and 21 d, and physiological traits, osmotic adjustment, oxidative status, antioxidant enzyme activities, and transcriptomic responses were systematically evaluated. Transcriptome profiling was performed using drought-treated and time-matched well-watered leaves with three biological replicates per group. Leaf relative water content and visible phenotypic changes remained relatively stable after 7 d but deteriorated markedly during prolonged drought. Soluble sugar accumulation increased from day 14, whereas MDA, O2−, and H2O2 accumulation became prominent mainly on day 21. Antioxidant enzymes exhibited distinct temporal patterns, with early activation of CAT, late reduction in POD activity, and delayed induction of SOD. Transcriptome analysis identified 18,550, 20,362, and 46,178 differentially expressed genes at 7, 14, and 21 d of drought treatment, respectively. A core set of 1530 genes responded consistently throughout drought progression, including 1253 upregulated and 277 downregulated genes. Shared upregulated genes were mainly associated with photosynthesis, carbon fixation, carbon metabolism, and DNA repair-related processes, while AP2/ERF, WRKY, and NAC were the predominant transcription factor families among the consistently upregulated genes. These findings reveal distinct temporal patterns of physiological and transcriptional responses in K. prostrata, characterized by changes in antioxidant enzyme activities, soluble sugar accumulation, and extensive transcriptional responses associated with oxidative stress under prolonged water withholding.

Plants doi: 10.3390/plants15223522

Authors: Xue Wang Xianhong Xu Jianqiang Huo Zhanmei Ma Rong Zhang

In the calcareous soils of Qinghai Province, phosphorus is easily fixed by calcium, restricting the growth and production of spring rapeseed. To clarify the root morphological characteristics of low-phosphorus-tolerant spring rapeseed in this area, a pot experiment was conducted using seven Qingza-series Brassica napus L. cultivars under P application (P1) and no-P (P0) treatments. Seedling biomass, phosphorus accumulation, root morphology, and vertical distribution were measured to evaluate the low-phosphorus tolerance of different varieties. Significant genotypic differences in low-phosphorus tolerance were observed among the varieties, with Qingza 12 (QZ12) showing the best tolerance, which was mainly achieved by maintaining large root surface area and intensive top-soil root aggregation for phosphorus capture under low-phosphorus conditions. Under low-phosphorus conditions, the relative aboveground biomass and total plant phosphorus accumulation were 47.31% and 46.07%, respectively, the root-crown ratio decreased but the total root length and root surface area remained maximal. More than 80% of the roots were concentrated in the 0–5 cm soil layer, with low-phosphorus conditions further enhancing surface aggregation. A low root-crown ratio, predominantly superficial roots, and strong surface aggregation can serve as seedling-stage identification criteria for low-phosphorus-tolerant spring rapeseed in Qinghai’s calcareous soils. QZ12 exhibits good low-phosphorus tolerance at the seedling stage and may act as a candidate germplasm resource for follow-up research in the Qinghai Province.

Plants doi: 10.3390/plants15223521

Authors: Lijun Qin Jinzhi Xu Xuemei Xian Lie Yu Xingyong Yang

The development of novel, efficient, and stable plant growth-promoting bacterial (PGPB) inoculants is crucial for mitigating environmental pollution and ensuring food security. Bacillus licheniformis GN02 has demonstrated significant plant growth-promoting effects. The final nitrogen content of the culture supernatant of GN02 grown in Ashby’s nitrogen-free medium reached 55.8 ± 1.92 mg/L. Genomic annotation of GN02 predicted genes for ammonium assimilation and nitrate reduction, but no canonical nifHDK nitrogenase genes. It also predicted plant growth-promoting potentials, including genes associated with siderophore, L-tryptophan, and polyamine biosynthesis, as well as motility and chemotaxis. Pot experiments on cotton seedlings were conducted with single and double inoculation of GN02 at 104 and 108 CFU/mL. The lower inoculation concentration (104 CFU/mL) exhibited a greater growth-promoting performance: it significantly increased indole-3-acetic acid, brassinosteroid, jasmonic acid, and salicylic acid concentrations and up-regulated YUCCA4, BES1, OPR3, ERF, and NPR1, alongside elevated rhizosphere urease and dehydrogenase activity and enriched root-associated genera such as Opitutus, Aquabacterium, and Hydrogenophaga. These findings demonstrate that GN02-mediated growth promotion is accompanied by root endosphere microbiome shifts and alterations in host hormone signaling. As a promising PGPB candidate, GN02 requires application strategies tailored to strain traits, with a suitable inoculation concentration and frequency.

Plants doi: 10.3390/plants15223520

Authors: Yulian Voynikov Teodor Marinov Paraskev Nedialkov

Phloroglucinol α-pyrones (PG α-pyr) are acylphloroglucinol–α-pyrone conjugates characteristic of Helichrysum and related genera. Helichrysum arenarium, Helichrysum stoechas and Achyrocline satureioides, three medicinal species not yet examined for the class, were analysed by UHPLC–Orbitrap–MS. The spectra were annotated by an automated R workflow that applies the fragmentation rules established for H. italicum to every MS2 spectrum and matches each against a dictionary of 527 expected compounds. The workflow annotated 175 PG α-pyr derivatives in the three samples (151 in H. arenarium, 53 in H. stoechas, and 32 in A. satureioides; 53 of them in more than one sample); with the 58 compounds published for H. italicum, the four species hold 189 PG α-pyr. Six were detected in all four: arzanol, 6-O-desmethylauricepyrone, 23-methyl-6-O-desmethylauricepyrone, 18,18-bis-desmethyl achyroclinopyrone A, a 3-non-alkylated pentanoyl derivative and helipyrone A. H. italicum and H. arenarium contain 153 and 91 µg/g of PG α-pyr (THAP equivalents), dominated by arzanol; H. stoechas (9.3 µg/g) and A. satureioides (3.2 µg/g) are each dominated by one compound, a dehydro-geranyl benzopyran and helipyrone A. Unsupervised clustering of the MS/MS spectra recovered the scaffold and the α-pyrone substituent. The class is common to the four samples; the dominant subclass and the preferred substituents distinguish them.

Plants doi: 10.3390/plants15223519

Authors: Desislava Mantovska Ana-Maria Nedelcheva Ginka Cholakova Alexandra Kapogianni Kalina Alipieva Svetlana Simova Zhenya Yordanova

Betonica bulgarica Degen & Nejceff is an endemic medicinal plant found only in Bulgaria. It is a valuable source of phenylethanoid glycosides and iridoids and is associated with antioxidant and anti-inflammatory activities. In the present study, we investigated whether phytochemical variations occurring during ex situ conservation of the species influence its immunomodulatory activity on pro-inflammatory response. Methanolic extracts obtained from in vitro-cultivated and ex vitro-adapted plants were evaluated for their effects on complement activity and on LPS-stimulated macrophages by assessing nitric oxide, TNF-α, IL-6, IL-12 production, and COX-2 expression. Comparative NMR metabolite profiling revealed the presence of verbascoside and the iridoids allobetonicoside and 8-O-acetylharpagide in in vitro- and ex vitro-adapted plants. Both variants exhibited anti-complement activity through binding to IgG-C1q complexes. The nitric oxide and TNF-α levels were reduced by extracts from in vitro-cultivated plants: 38.68 μM NO2 and 208.64 pg/mL, respectively, whereas extracts from ex vitro plants showed stronger suppression of IL-6 (168 pg/mL). None of the variants had an effect on IL-12 production. COX-2 expression was slightly reduced by the extract of in vitro-adapted plants. Overall, these findings demonstrate that phytochemical alterations associated with environmental adaptation during ex situ conservation influence the immunomodulatory activity of B. bulgarica, highlighting the dynamic relationship between plant metabolism and biological activity.

Plants doi: 10.3390/plants15223518

Authors: Myung Suk Ahn Dae-Won Ki Hyun Hwan Jung Jiwon Moon Yun-Hee Shin Bong-Sik Yun

Comparative information on the chemical and odor characteristics of absolutes from Aster species remains limited. This study compared the yield, volatile composition, and odor characteristics of absolutes prepared from the dried aerial parts of eleven Aster species native to Korea. Odor intensity and electronic-nose responses were evaluated, and volatile compounds were characterized by headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GC-MS) and multivariate analyses. Absolute yields ranged from 0.95% in A. ageratoides to 1.48% in A. danyangensis, whereas odor intensity was highest in A. indicus (1712 A.U. µL−1). A total of 150 volatile compounds were identified, and sesquiterpene hydrocarbons were the predominant chemical class in ten of the eleven species. Hierarchical clustering separated the species into four compositional groups, with A. koraiensis forming a distinct cluster characterized by high relative contents of β-sesquiphellandrene and β-bisabolene. Electronic-nose profiles differed among some species but did not distinguish A. koraiensis from A. pseudoglehnii and A. incisus. Woody, Spicy, Herbal, and Balsamic were the major database-derived odor classes. Overall, the absolutes differed among species in both volatile composition and odor characteristics, and these results provide a basis for comparing native Aster species as natural fragrance materials.

Plants doi: 10.3390/plants15223517

Authors: Chandrasekhar Manikala Phuwadol Bangrak Thanet Khomphet Siti Nur Aisyah

Understanding how soil nutrient availability translates into leaf nutritional status, fruit-water mineral composition and reproductive performance is important for improving nutrient management of aromatic coconut. This study evaluated these relationships in aromatic coconut across two contrasting locations in southern Thailand. Two coconut varieties were assessed for fruit-water composition, reproductive performance, soil nutrient availability, and leaf mineral composition. Fruit-water Ca concentration was significantly higher in Trang, while Mg concentration was significantly higher in Var1 than in Var2. Fruit-water K and Na showed no significant differences between locations or varieties. Fruit-water pH was also higher in Trang, whereas TSS did not differ significantly. Bunch number and number of inflorescences were higher in Trang, while Var2 produced more female flowers per bunch and more fruits per palm. Soil P, K, Ca, Mg, and Na were substantially higher in Nakhon Si Thammarat than in Trang, but these differences were not consistently reflected in leaf nutrient concentrations. Soil Ca and K showed significant but modest relationships with their corresponding leaf nutrient concentrations, whereas Mg and Na showed no significant predictive relationships. Leaf nutrients were strongly interrelated, but their relationships with fruit minerals, TSS and reproductive traits were generally weak after accounting for location. PCA clearly separated the two locations, with PC1 and PC2 explaining 29.0% and 17.8% of the total variance, respectively, while the first four PCs accounted for 67.2%. Soil Mg and Na and leaf K, Ca, and Mg were among the major contributors to this multivariate differentiation. Nutrient relationships in aromatic coconut were nutrient-specific and strongly environment-dependent, rather than representing simple soil-to-leaf-to-fruit transfer. Integrated soil and leaf nutritional assessment may therefore provide a more reliable basis for coconut nutrient management than soil analysis alone. The findings highlight the importance of considering both soil and leaf nutrient status when evaluating nutritional variation in aromatic coconuts, while validation across multiple seasons and environments is needed before broader nutritional or agronomic recommendations can be made.

Plants doi: 10.3390/plants15223516

Authors: Peipei Li Xinyue Han Fupeng Wei Caiwen Qi Xuetong Zhao Zimeng Wang Xiaoya Niu Tiantian Liu Enqi Yang Yemeng Zhang

Trehalose-6-phosphate phosphatase (TPP) is a key enzyme in trehalose biosynthesis and participates in endogenous sugar signaling regulation and abiotic stress adaptation in plants. We performed comprehensive bioinformatic analyses of the CqTPP gene family in Chenopodium quinoa, including physicochemical property evaluation, phylogenetic analysis, conserved motif identification, gene structure characterization, chromosomal localization, synteny analysis, and cis-acting elements prediction. In total, 12 CqTPP genes were identified and grouped into three distinct subgroups with obvious divergence in conserved motifs and gene structures. Segmental duplication dominated the expansion of the CqTPP gene family, and duplicated gene pairs underwent strong purifying selection. CqTPP genes shared high sequence homology with homologs from Beta vulgaris and Spinacia oleracea, and their promoters contained abundant stress-, hormone-, and light-responsive cis-acting elements. Transcriptome data revealed divergent salt-responsive expression patterns. Notably, CqTPP10 was persistently and significantly upregulated with prolonged salt treatment. Heterologous overexpression of CqTPP10 in Arabidopsis thaliana enhanced salt tolerance, improved seed germination, elevated SOD and POD activities, increased trehalose levels, reduced electrolyte leakage, reduced MDA and H2O2 accumulation, and downregulated KIN10. Collectively, this work provides comprehensive insight into the evolution and salt-response features of the quinoa CqTPP gene family and supplies promising candidate genetic resources for salt-tolerance improvement in crops.

Plants doi: 10.3390/plants15223514

Authors: Yu-Long Li Jia-Yi Chen Meiru Yang Qiong Yu

Glucosinolates (GSLs) are sulfur- and nitrogen-containing specialized metabolites characteristic of the Brassicales, especially Brassicaceae crops. The agricultural and nutritional significance of GSLs is not determined by total content alone. Rather, it reflects the combined effects of chemical diversity, organ-specific accumulation, and hydrolytic conversion. Biosynthesis shapes the classes and side-chain structures of GSLs, transport regulates their allocation to different tissues, and myrosinase-mediated hydrolysis generates bioactive products. These coordinated processes ultimately affect plant defense, crop flavor, nutritional quality, and potential toxicity. In this review, we systematically summarize the biosynthetic pathways, hydrolysis mechanisms, and transport regulation of GSLs, emphasizing how these mechanisms jointly determine their biological functions and application potential. We further discuss the roles of GSLs in crop defense, rapeseed feed quality, vegetable flavor formation, functional food development, and food safety evaluation. Rather than pursuing a simple increase or decrease in total GSL levels, future GSL regulation should focus on crop-, tissue-, compound-, and product-specific strategies. Multi-gene regulatory networks, tissue-specific allocation, and product-specific safety assessment will be essential for balancing plant defense, agricultural productivity, food quality, and human health benefits.

Plants doi: 10.3390/plants15223515

Authors: Qianyun Jin Rongwan Zhang Chenglei Zhong Luyi Zhang Yinuo Chen Yanan Jiang Hemin Yang Shuting Li Sushu Yang Zhenghao Fu Jiangmin Xu Song Yan Kongjian Wang Yuchun Rao

Rice (Oryza sativa L.) is a major staple crop in China, but high-temperature heat stress has become one of the key factors threatening national food security, causing symptoms such as leaf curling, chlorosis, and water loss-induced wilting. In this study, we used indica rice ‘HZ’ (Oryza sativa L. subsp. indica cv. ’HZ’), japonica rice ‘Nekken2’ (Oryza sativa L. subsp. japonica cv. ’Nekken2’), and their derived 120 recombinant inbred lines (RILs) as experimental materials. Phenotypic data including seedling survival rate, heat tolerance rating, and seedling growth rate were measured, and quantitative trait locis (QTLs) associated with rice seedling heat tolerance were mapped using a high-density genetic map. A total of 31 QTLs related to heat tolerance were detected, with logarithm of odds (LOD) values ranging from 2.06 to 5.09. Additionally, 12 candidate genes were identified within these QTL intervals. Real-time quantitative reverse transcription PCR (qRT-PCR) analysis revealed that LOC_Os03g45330, LOC_Os04g32560, and LOC_Os10g26720 associated with rice heat tolerance, whereas LOC_Os01g46080 and LOC_Os07g08340 may contribute to it. These findings provide important insights for further elucidating the molecular mechanisms underlying heat stress response in rice and for breeding new high-quality heat-tolerant rice varieties.

Plants doi: 10.3390/plants15223513

Authors: Jianwei Zhang Rong Ning Jiaxiang Luo Liwei Bai Fenglian Wu Qin Yang Jianwen He Huanxiu Li

Low temperature is a major abiotic stress that constrains the early-spring and late-season cultivation of pepper (Capsicum annuum L.). Basic helix–loop–helix (bHLH) transcription factors play important roles in plant stress responses; however, their functions in pepper cold tolerance remain poorly understood. In this study, we characterized CabHLH55 and investigated its role in cold stress through bioinformatic analysis, subcellular localization, quantitative reverse transcription PCR (RT-qPCR), heterologous overexpression in Arabidopsis, and virus-induced gene silencing (VIGS) in pepper. CabHLH55 encodes a 257-amino-acid nuclear protein containing a conserved bHLH domain. Its promoter contains multiple cis-acting elements associated with phytohormone and stress responses. Expression analysis showed that CabHLH55 responded to multiple abiotic stresses and was markedly downregulated under cold stress. Under cold conditions, CabHLH55-overexpressing Arabidopsis plants exhibited reduced chlorophyll content and increased relative electrolyte leakage and malondialdehyde (MDA) levels compared with WT plants, indicating enhanced cold-induced damage. Conversely, silencing CabHLH55 in pepper alleviated cold injury, reduced reactive oxygen species (ROS) accumulation and MDA levels, and increased peroxidase (POD) activity. Moreover, the expression of the cold-responsive genes CaCBF1a, CaKIN, and CaCOR47 was significantly increased in CabHLH55-silenced plants under cold stress. Collectively, these results indicate that CabHLH55 functions as a negative regulator of cold tolerance in pepper, potentially by affecting ROS homeostasis and cold-responsive gene expression. This study provides a basis for further elucidating bHLH-mediated cold stress responses in pepper and may facilitate the molecular breeding of cold-tolerant pepper cultivars.

Plants doi: 10.3390/plants15223509

Authors: Sumin Lv Penglei Yan Xuan Li Ziyi Yi Fu Zhou Xiao Zhang Nannan Zhang

Cotton Verticillium wilt and nitrogen deficiency can produce similar chlorotic symptoms and spectral responses, while variation among growth stages may further limit model transferability. We used leaf-level hyperspectral and SPAD data from FS, BS, and BO to classify three operational classes—healthy-reference (H), nitrogen-deficiency (N), and Verticillium-wilt (W)—and assess preprocessing, feature selection, classification, and cross-stage transfer. N and W were defined by reduced nitrogen supply and characteristic field symptoms, respectively, without independent tissue-nitrogen or pathogen confirmation. Random forest–recursive feature elimination (RF-RFE) and a mutual-information-based particle swarm optimization filter (Filter-PSO) each selected 20 spectral bands, and support vector machine (SVM), Light Gradient Boosting Machine (LightGBM), and Extreme Gradient Boosting (XGBoost) were integrated into an Opt-SLX probability-fusion ensemble. Cross-stage analyses compared direct transfer with correlation alignment (CORAL) and mean–variance alignment. Repeated cross-validation on the development set showed that SG-MSC increased mean overall accuracy (OA) relative to raw spectra by 28.58, 27.78, and 35.71 percentage points at FS, BS, and BO, respectively. Under the fixed 20-band setting, classifier and ensemble performance depended on both growth stage and feature-selection pipeline. RF-RFE–Opt-SLX achieved held-out test OA values of 0.9206, 0.9286, and 0.9127 at FS, BS, and BO, respectively, whereas RF-RFE–SVM achieved the highest OA at BS (0.9365). Direct cross-stage transfer declined markedly, with mean OA ranging from 0.4206 to 0.5556 under RF-RFE and from 0.3968 to 0.5159 under Filter-PSO; transfer performance showed no simple monotonic relationship with the calendar interval between stages. Domain adaptation increased performance in some transfer tasks, but the magnitude and direction of the gains depended on the specific transfer task and feature representation. These results demonstrate pronounced growth-stage dependence in spectral discrimination among the H, N, and W operational classes and show that high within-stage classification accuracy does not necessarily translate into robust cross-stage transferability. Independent validation incorporating tissue nitrogen measurements, pathogen confirmation, and multi-environment datasets is needed to establish model robustness and generalizability.

Plants doi: 10.3390/plants15223511

Authors: Irina Ielciu Ana Maria Vlase Daniela Hanganu Emoke Pall Andrei Mocan Mihai Babotă Oleg Frumuzachi Laurian Vlase Gianina Crișan Ramona Păltinean

Background/Objectives: Corydalis cava (L.) Schweigg. & Körte and Corydalis solida (L.) Clairv. are perennial geophytes belonging to the Papaveraceae family. The present study investigated the phytochemical composition, antimicrobial activity and cytotoxic effects of their hydroethanolic extracts. Methods: The phenolic profiles of aerial parts were characterized by spectrophotometric assays, and individual polyphenolic compounds were identified and quantified by LC-HRMS/MS. Antioxidant activity was assessed using the TEAC and EPR assays. The cytotoxic effects were evaluated on DLD-1 human colorectal adenocarcinoma cells and HFL human lung fibroblasts. The antimicrobial effects were assessed against a selection of Gram-positive and Gram-negative bacterial strains. Results: The results showed high amounts of flavonoids and polyphenols, but also of individual metabolites such as phenolic acids, flavonol aglycones and their glycosides. Cytotoxicity assays demonstrated good tolerability towards normal cells, indicating inhibitory effects only on tumor-derived cell lines. In addition, the tested samples exhibited strain-dependent antimicrobial activity. Conclusions: Collectively, the results obtained hereby suggest that C. cava and C. solida represent a potential source of bioactive metabolites, with promising antimicrobial, antioxidant and cytotoxic bioactivities.

Plants doi: 10.3390/plants15223512

Authors: Jie Wang Lijun Zhu Houji Liu Zhikun Yang Juntuan Zhai Yongfu Chai Zhijun Li

Dryland riparian forests depend on river-derived water in arid landscapes, but the relative associations of regional climate, local soil moisture, and biodiversity with ecosystem multifunctionality (EMF) remain uncertain. We analysed data from 516 plots in Populus euphratica forests across northwestern China. EMF was quantified from eight vegetation and soil attributes using averaging and multi-threshold approaches. We combined species richness, Rao’s quadratic entropy, community-weighted trait axes, climate principal components, and soil principal components in mixed-effects models, hierarchical partitioning and partial least-squares path models. EMF declined from near- to far-riparian plots, whereas soil water content did not differ significantly among distance bands. Hydroclimatic stress was negatively associated with EMF, especially in far-riparian plots and at higher thresholds, while soil moisture was positively associated mainly at low-to-intermediate thresholds. The relative explanatory contributions of biodiversity varied across riparian positions and predefined functional thresholds: RaoQ contributed most near rivers at lower thresholds, species richness was more important at intermediate and far positions, and community-weighted traits became prominent under selected high-threshold conditions. Species richness and community-weighted trait axes also changed with riparian distance, whereas RaoQ was comparatively stable. In the PLS-PMs, modelled direct abiotic associations were generally stronger than indirect associations involving biodiversity. Given the observational study design and correlational analyses, these findings indicate context-dependent associations among climate, soil moisture, biodiversity and ecosystem multifunctionality rather than causal relationships.

Plants doi: 10.3390/plants15223510

Authors: Zlatko Liber Zlatko Šatović Boštjan Surina Toni Nikolić Filip Varga Martina Grdiša

Isolated mountain ecosystems provide an opportunity to study how long-term natural fragmentation shapes population persistence. Unlike species fragmented by human activities, many mountain endemics have evolved under naturally discontinuous conditions over long timescales, allowing historical and contemporary processes to be disentangled. Degenia velebitica (Brassicaceae) represents a suitable model for such studies. We analysed nuclear microsatellites, chloroplast DNA, fine-scale spatial genetic structure, effective population size, and demographic characteristics across all currently known populations. Despite pronounced genetic differentiation, restricted contemporary gene flow, small effective population sizes, and marked demographic differences, populations maintained moderate nuclear genetic diversity. Chloroplast DNA revealed low haplotype variation and strong spatial structuring of maternal lineages, consistent with limited historical seed-mediated dispersal. Importantly, demographic and genetic indicators were not always concordant: in one population, an apparent recent decline in census size and low effective population size were accompanied by relatively well-preserved nuclear genetic diversity. These findings demonstrate that neutral genetic diversity alone may provide an incomplete picture of population status and evolutionary potential, highlighting the importance of integrating genetic, demographic, and ecological information and maintaining existing population structure and habitat integrity in conservation planning.

Plants doi: 10.3390/plants15223506

Authors: Li Cao Liping Tang Gilles Colinet Lin Chen Yumo Duan Yongmei He Na Jiang Xinran Liang Yanqun Zu Zuran Li

Elucidating the transfer and accumulation mechanisms of Cd in wild rice resources is crucial for breeding low-Cd rice varieties. This study investigated the physiological responses and Cd distribution characteristics of Oryza meyeriana Baill. under 5 and 15 mg/kg Cd treatments. A pot experiment was conducted using O. meyeriana, along with the low-Cd and high-Cd cultivar for comparison. Scanning electron microscopy–energy-dispersive X-ray spectroscopy (SEM-EDX) and non-invasive micro-test technology (NMT) were employed to investigate Cd transfer in roots. Under Cd stress, O. meyeriana adopted morphological adaptations, including decreased root hair density and length, alongside a thickened root endodermis. NMT revealed a net Cd2+ influx at the root surface that intensified with increases in external Cd concentration, with negligible root efflux. Combined with the tissue Cd distribution gradient (root > stem > leaf), these physiological kinetics demonstrate that O. meyeriana has a prominent root Cd retention strategy, with roots sequestering 52–59% of total plant Cd. O. meyeriana exhibited lower Cd translocation in both the xylem and phloem. Furthermore, Cd excretion was detected on the leaf surface of O. meyeriana, functioning as an additional detoxification pathway, with excretion amounts reaching 13.77 μg/kg under 15 mg/kg Cd treatment. O. meyeriana exhibits physiological sensitivity to Cd but copes with the stress through distinct morphological adaptations. Collectively, O. meyeriana responds to Cd stress through a coordinated regulatory pathway of root retention—restricted stem to leaf translocation—and leaf excretion, thereby exhibiting strong low Cd accumulation capacity and highlighting its valuable application potential in low-Cd rice breeding.

Plants doi: 10.3390/plants15223508

Authors: Tatiana Leonova Elena Tsvetkova Anastasia Aslanova Anastasia Orlova Svetlana Silinskaia Alisa Sokolova Ludger A. Wessjohann Andrej Frolov Anastasia Kurina

St. John’s wort is a medicinal plant of significant pharmaceutical importance, yet its metabolic composition and biological activity vary considerably depending on genetic and environmental factors. Understanding the interplay between genotype and growing conditions is essential for ensuring reproducible therapeutic properties of herbal raw materials. This study aimed to comprehensively evaluate the effects of genotype and growing location on the metabolite profiles and antibacterial/antioxidant activities of H. perforatum. Seven accessions from the VIR collection were grown in two climatically distinct regions of Russia: the temperate zone (Pushkin, Leningrad Region) and a warm region (Maykop, Republic of Adygea). Methanolic extracts were analyzed using LC-MS/MS, yielding 89 annotated metabolites belonging to 8 major compound classes. Antibacterial activity was assessed against Gram-positive (Staphylococcus aureus, Micrococcus luteus) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) strains, while antioxidant capacity was evaluated using DPPH and TEAC assays. PCA identified cultivation location (36.7%) and genotype (14.8%) as the major sources of metabolic variation, with a significant location × cultivar interaction detected for 70.2% of all analysed features. Naphthodianthrones and procyanidins consistently accumulated in the shoots of H. perforatum plants grown in the temperate region, while flavonoids and acylphloroglucinols exhibited genotype-dependent responses. Extracts from the plants grown in Pushkin showed more pronounced antibacterial activity, particularly against M. luteus and E. coli, and higher antioxidant capacity compared to the Maykop-grown plants, correlating with elevated levels of bioactive compounds. Genotype k-14 displayed the most pronounced metabolic plasticity. The metabolite profiles and bioactivity of H. perforatum are associated with complex genotype-environment interactions. These findings highlight the importance of considering both genetic background and cultivation location for the production of standardized herbal materials with more consistent pharmacological properties.

Plants doi: 10.3390/plants15223507

Authors: Longfei Li Lijuan Gao Jintao Xu Huan Liu Yue Yao Gang Niu Xinfang Zhang Minghui Ji Baofeng Hao

Evaluation of a selected fruit cultivar requires identifying its distinctive combination of traits relative to relevant breeding materials. We evaluated the pear cultivar Suyu (SY) against its parents Mili (ML) and Dangshansuli (DS) using fruit quality measurements, descriptive sensory scores, texture profile analysis, electronic nose responses and headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME-GC-MS) volatile signals. Parent-referenced trait contrasts were combined with blockwise multivariate comparisons and sensitivity analyses of scaling and feature inclusion. In the sampled batches, SY had a mean soluble solids content of 14.10%, exceeding ML and DS by 20.4% and 11.2%, respectively, and had greater measured hardness than both parents. Its soluble sugar content was lower than ML, showing that the observed contrast was trait-dependent. Under unit-variance scaling, SY–DS to SY–ML centroid-distance ratios were 0.623 for fruit quality, 0.269 for texture, 0.837 for electronic nose responses and 0.821 for volatile profiles. Within this single-site, single-year dataset, the direction of closer resemblance to DS persisted under range scaling and single-feature omission, and after excluding specified ratio or composite traits. Of 656 distinct numerical volatile profiles, 58 met the exploratory screening criteria for SY versus ML and 40 for SY versus DS, with 17 shared. Under the examined analytical choices, descriptive resemblance to DS coexisted with distinctive soluble-solids, mechanical and volatile-signal attributes. This parent-referenced evaluation identifies traits and signals for validation in larger cultivar sets and replicated field trials; it does not estimate parental genetic contributions or establish consumer preference. Maturity was judged qualitatively, and storage duration varied with harvest date. The contrasts therefore describe the sampled fruit batches, with additional uncertainty from tree clustering and the small number of pools.

Plants doi: 10.3390/plants15223505

Authors: Junling Li Mengxia Li Xian Xue Xiaofei Wang Haotian Ye Zhihong Zhang

Flowering-stage heat and water limitation are important environmental constraints for summer maize, but their co-occurrence and remotely sensed expression vary with the phenological timing and the indicators used. We divided the Huang–Huai–Hai Plain into six phenological regions with fixed representative flowering windows and integrated ERA5-Land air temperature and 0–20 cm soil moisture for 1980–2025, MODIS variables for 2002–2025, a 2025 field survey, and long-term agricultural station records. The hourly heat exposure was calculated within each region-specific 10-day window after converting ERA5-Land timestamps from UTC to China Standard Time. Two-way fixed-effects models with 1° spatial-block clustered standard errors were used for the ERA5–MODIS panel. The mean flowering exposures were 22.09 h above 32 °C and 2.71 h above 35 °C; both the H32 and H35 increased significantly, whereas the soil moisture deficit indices showed no significant trend. Concurrent flowering heat and soil moisture deficit accounted for 13.56% of weighted grid-years, compared with 6.45% expected under independence from the pooled marginal frequencies. Antecedent soil moisture deficit occurred in 60.50% of concurrent grid-years, compared with 43.14% of flowering-deficit-only grid-years. In the state fixed-effects models, concurrent stress was associated with higher land surface temperature and lower NDVI, LAI, and evapotranspiration values. However, joint models containing heat, soil moisture deficit, and their interaction provided little evidence of a general multiplicative interaction. In the full 2025 survey, the flowering soil moisture deficit, concurrent index, and daytime land surface temperature were negatively associated with kernel number. None of these associations remained FDR-significant in the Region 2 (R2)-only sensitivity analysis, indicating that the full-survey associations could not be clearly separated from regional differences. Long-term station models did not detect FDR-significant associations between the environmental-stress indices and yield components, but seven flowering or post-flowering MODIS indicators were associated with theoretical-yield anomalies after FDR correction. These findings support phenology-aligned regional monitoring, while the reported relationships remain observational.

Plants doi: 10.3390/plants15223504

Authors: Madira C. Manganyi Olawole E. Aina Jeffrey Lebepe Nana M. D. Buthelezi Christ Donald Kaptchouang Tchatchouang

Climate change, soil degradation, water scarcity and increasing food demand are intensifying the need for sustainable strategies that improve crop productivity and resilience while reducing chemical inputs. Beneficial bacterial and fungal endophytes associated with plant roots are promising next-generation plant biostimulants because they colonize internal plant tissues and modulate nutrient acquisition, phytohormone signalling, metabolism and stress responses. This review focuses specifically on beneficial bacterial and fungal endophytes, particularly root-associated endophytes, and examines their roles in plant growth promotion, nutrient mobilisation, and tolerance to abiotic and biotic stresses. Emphasis is placed on microbial colonisation, molecular signalling and emerging multi-omics approaches for elucidating endophyte–plant interactions. Pathogenic endophytes and seed-specific endophyte systems are outside the primary scope of this review. Despite promising mechanistic evidence, translation to field applications remains constrained by host specificity, environmental variability, microbial compatibility, formulation and delivery, persistence, biosafety and regulatory requirements. Future development should therefore move beyond the selection of individual strains towards predictive, mechanism-based and microbiome-informed strategies supported by multi-omics and rigorous field validation. Such approaches could enable reliable, context-adapted endophyte-based biostimulants for sustainable and climate-resilient agriculture.

Plants doi: 10.3390/plants15223503

Authors: Martina Orlando-Bonaca Ana Lokovšek Valentina Pitacco

Canopy-forming macroalgae, particularly Cystoseira sensu lato species, are key habitat-forming organisms and important indicators of ecological status (ES) in Mediterranean coastal ecosystems. We assessed long-term changes in infralittoral macroalgal assemblages along the Slovenian coastline (northern Adriatic) using data from the national monitoring programme. Spring and autumn surveys conducted between 2007 and 2025 at 13 stations across four coastal water bodies were analysed using the Ecological Evaluation Index continuous formula (EEI-c) and complementary structural and functional indicators. Temporal trends were tested with Spearman rank correlations. Total macrophyte coverage, the relative coverage and taxonomic proportion of canopy-forming taxa, and the relative coverage of Ecological State Group I (ESG I) taxa declined significantly in both seasons. In contrast, the proportion of ESG I taxa in total taxonomic richness showed no significant temporal trend. Autumn assemblages generally had higher total coverage and relative ESG I coverage, whereas the relative canopy contribution was generally higher in spring. Seasonal differences therefore accompanied a decline detectable in both seasonal series. Stable ESG I taxonomic ratios did not imply stable coverage or habitat structure. These results support seasonally replicated monitoring and conservation of remaining macroalgal forests. Identifying the drivers of decline requires environmental measurements alongside biological surveys.

Plants doi: 10.3390/plants15223501

Authors: Muhammad Nadeem Abbas Mengqin Chen Saima Kausar Huixue Chen Hongjuan Cui

Phytosterols are plant-derived sterols that are structurally similar to cholesterol and have been extensively studied for their biological roles in lipid metabolism. Emerging evidence also suggests that they may have an effect on inflammatory responses. Here, we focus on the membrane-related functions of phytosterols, with specific emphasis on how structural differences modify membrane organization and physical properties, and thereby influence inflammatory activities. In model membranes, stigmasterol, campesterol and β-sitosterol exhibit different effects on lipid packing, membrane order and physical properties in a structure-dependent manner. However, there is limited evidence that these changes are associated with inflammatory signalling. The key molecular evidence implies a relationship exists between p67 phox recruitment, NADPH oxidase activity, reactive oxygen species production and NLRP3-related responses, and PS-associated modifications in membrane lipid packing. Other cellular studies demonstrate alterations in prostaglandin release and inflammatory signalling, but lack biophysical properties of the membrane. Furthermore, phytosterol effects are not invariably anti-inflammatory and depend on the kind of sterol and the experimental conditions. Phytosterols may also modulate intracellular pathways linking inflammatory responses with cellular lipid homeostasis, such as LXR/ABCA1 and MAPK signalling. The extent to which membrane remodelling contributes to these effects is unknown, and further investigation is required to determine whether the proposed mechanism applies to other phytosterols and experimental models. Further studies are needed, particularly with the incorporation of phytosterols into immune-cell membranes in vivo and the relative contributions of membrane effects and downstream metabolic processes.

Plants doi: 10.3390/plants15223502

Authors: Fahadul Islam Rafiuddin Khan Labib Mehrukh Zehravi Mashia Subha Lami Rajib Das Laliteshwar Pratap Singh Jithendar Reddy Mandhadi P. Balan Jishan Khan Sharuk L. Khan Firzan Nainu Mohamed H. Nafady Safia Obaidur Rab Talha Bin Emran Polrat Wilairatana

In the published publication [...]

Plants doi: 10.3390/plants15223500

Authors: Xi Sun Changbin Niu Zhenrong Qiu Honglin Huang Shangfeng Zhou Haona Yang Ke Chen Di Wu Long Wang Chunhui Zhang Feng Yu Lifeng Wang Changsheng Li

Allelopathy mediates below-ground competitive interactions between rice and weeds in paddy ecosystems. Diverse phytotoxic allelochemicals complicate their simultaneous quantification in root exudates. This study aimed to develop a targeted LC-MS/MS method for profiling fourteen allelochemicals spanning five classes. Multiple reaction monitoring (MRM) transitions were optimized for reference standards, and method performance was validated for linearity, sensitivity, precision, recovery, and matrix effects. All analytes exhibited good linearity (R2 > 0.99), with limits of detection (LODs) of 0.005–0.641 ng/mL and limits of quantification (LOQs) of 0.017–2.136 ng/mL. Intra- and inter-day precision ranged from 0.27–4.90% and 0.45–4.51%, respectively, while matrix effects were 86.21–113.99%. In the root exudates of rice, Echinochloa crus-galli, Leptochloa chinensis, and sorghum, caffeic acid, p-hydroxybenzoic acid, syringic acid, vanillic acid, loliolide, and DMBQ were detected in all species, whereas the other analytes exhibited species-specific distribution patterns. Notably, momilactone A was only detected in Nipponbare (NIP) rice and E. crus-galli root exudates. Principal component analysis (PCA) based on 11 detectable allelochemicals revealed distinct profiles among the four hydroponically cultured species. This validated LC-MS/MS platform enables characterization of allelochemical compositions in paddy root exudates.

Plants doi: 10.3390/plants15223499

Authors: Waqas Liaqat Adil Bakoğlu Muhammed İkbal Çatal Hamza-Ali Khan Muhammad Tanveer Altaf

Optimizing harvest timing and genotype selection is essential for forage quality in grass pea (Lathyrus sativus L.). Twelve genotypes were evaluated at five weekly harvest timings in a one-site, one-season (2019) field trial in Rize, on the humid Black Sea coast of Türkiye. Crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), acid detergent protein (ADP), P, K, Ca, and Mg were predicted by near-infrared reflectance spectroscopy (NIRS) on one sample per genotype × harvest combination, and digestibility and energy indices were calculated from ADF and NDF. Harvest timing affected all variables (p < 0.01). With later harvests, ADF and NDF generally increased, CP declined after peaking at the third harvest (29.07%), and calculated digestible dry matter and relative feed value decreased. The first three harvest timings (vegetative growth to early-to-mid flowering) combined higher CP with lower fiber. Genotypes differed in CP, P, K, Ca, and mineral ratios (p < 0.05) but not in fiber fractions or calculated indices. Because the genotype × harvest interaction could not be tested and biomass yield and β-ODAP were not measured, genotype comparisons are provisional. Fiber accumulation was associated with declining forage quality, highlighting the importance of early harvest timing.

Plants doi: 10.3390/plants15223498

Authors: Shehnaz Kousar Sheraz Ahmad Johar Jamil Sumaira Salahuddin Lodhi Satyabrata Nanda Rahat Sharif Yuzhou Zhang Lixin Zhang

Alfalfa (Medicago sativa L.) is a major perennial forage legume, but its cultivation is increasingly limited by soil salinization and low temperature. YABBY proteins are plant-specific transcription factors best known for lateral organ development, although their role in abiotic stress adaptation remains unclear. In this study, eight MsYABBY members, encoded by seven loci, were identified in the alfalfa genome; seven were anchored to four chromosomes (Chr2, Chr4, Chr5 and Chr8) and one to an unanchored scaffold. Phylogenetic analysis with Arabidopsis and rice YABBY proteins classified the MsYABBY members into the five canonical YABBY subfamilies (FIL/YAB3, YAB2, YAB5, INO and CRC). Transcriptome and qRT-PCR profiling further revealed stress-dependent expression, among which MsYABBY8 was most strongly induced under salt and cold stress. Subcellular localization confirmed that the MsYABBY8 protein was localized to the nucleus. To further validate its function, MsYABBY8 was heterologously overexpressed in Arabidopsis, resulting in contrasting phenotypes under the two stresses. Under salt stress, the transgenic lines were more sensitive than the wild type, with reduced chlorophyll, survival, and relative water content, but increased thiobarbituric acid-reactive substances (TBARS), electrolyte leakage, and the Na+/K+ ratio. In contrast, the same lines performed better under cold stress, with higher chlorophyll content, photosynthetic rate, and antioxidant enzyme activities, and elevated AtCBF1 and AtCOR15A expression. Overall, these results obtained in a heterologous system suggest that MsYABBY8 may act as a negative regulator of salt tolerance but a positive regulator of cold tolerance, which remains to be confirmed in alfalfa.

Plants doi: 10.3390/plants15213497

Authors: Xiao Zhou Xingge Huang Shengyu Guo Yunzhu Chen Peiwang Li Qiang Liu Changzhu Li Mei Chen Wenbin Zeng Yanju Song Jingzhen Chen Yan Yang

Bud-bearing stem segments of citral-rich Camphora officinarum were used to optimize in vitro shoot proliferation and adventitious root induction by screening basal media and plant growth regulator types and concentrations. Physiological and endogenous hormonal responses were compared between the two culture stages. At each stage, plantlets cultured on the selected medium and a control medium without plant growth regulators were sampled on days 0, 7, 14, 21, and 28. Soluble sugar, soluble protein, and total phenolic contents; superoxide dismutase (SOD) and peroxidase (POD) levels; and zeatin riboside (ZR), gibberellic acid (GA3), indole-3-acetic acid (IAA), and abscisic acid (ABA) contents were determined. These parameters were also compared among medium formulations. Murashige and Skoog (MS) medium supplemented with 2.00 mg/L 6-benzyladenine (6-BA), 0.20 mg/L 1-naphthaleneacetic acid (NAA), and 0.05 mg/L kinetin (KT) yielded a proliferation coefficient of 8.16. Within the concentration ranges tested, 6-BA had a greater effect on proliferation than KT or NAA. Half-strength MS medium (1/2MS) was suitable for rooting, and 0.50 mg/L indole-3-butyric acid (IBA) produced a rooting rate of 92.22%. Under the selected culture conditions, soluble sugar content generally increased during shoot proliferation, soluble protein content peaked on day 14, and SOD and POD levels peaked on day 21. During rooting, soluble sugar content declined after day 7, whereas soluble protein content and SOD levels reached high values on day 14. During the middle and later culture periods, treated plantlets generally had higher IAA and lower ABA contents than the controls at both stages. Comparisons among formulations showed that the treatment favorable for proliferation had relatively high soluble sugar and ZR contents, whereas the treatment favorable for rooting had relatively low contents of both. This study identified suitable proliferation and rooting conditions for the genotype represented by the single donor tree examined and revealed differences in sugar dynamics and endogenous hormonal responses between the two culture stages. The selected growth regulator concentrations were suitable within the tested ranges for this genotype. Their applicability to other C. officinarum genotypes requires further validation.

Plants doi: 10.3390/plants15213496

Authors: Daju Chen Wenqing Zhang Tuo Zeng Caiyun Wang

Tanacetum coccineum is an ornamental plant capable of producing pyrethrins, natural insecticidal metabolites. Salinity restricts plant growth, but its effects on stress responses and pyrethrin-related metabolism remain poorly understood in this species. In this study, seedlings were treated with 0, 100, or 200 mM NaCl for 7 days, followed by physiological measurements and transcriptome analysis of leaf and root. Salt stress caused progressively more severe leaf injury and distinct antioxidant responses in the two tissues. Transcriptome analysis revealed extensive tissue-dependent transcriptional changes, including differential expression of genes associated with antioxidant defense, ABA biosynthesis, and pyrethrin metabolism. Several pyrethrin biosynthesis-related genes were upregulated under moderate salinity, whereas their expression patterns became more variable at higher NaCl concentrations. Meanwhile, spectrophotometrically estimated leaf pyrethrin content increased with NaCl concentration. WGCNA and PPI network analyses identified candidate hub genes associated with ion homeostasis, oxidative defense and carbon metabolism. Further analysis identified 19 candidate transcription factors and significant co-expression associations with ABA- and pyrethrin-related genes, suggesting potential transcriptional links between salt-stress responses and specialized metabolism. Collectively, these findings reveal tissue-dependent responses to salinity and provide insights into the transcriptional complexity of pyrethrin-related metabolism in T. coccineum.

Plants doi: 10.3390/plants15213495

Authors: Raires Irlenizia da Silva Freire Agda Malany Forte de Oliveira Renata Ramayane Torquato Oliveira David Emanoel Gomes da Silva Matheus Augusto Silva Daise Feitoza da Rocha Joilma Maria de Souza Elisandra Sampaio de Freitas Anderson Rodrigo Luciano da Silva Jéssica Crhistie de Castro Granjeiro Oliveira Lucas Rodrigues Bezerra da Silva Adriano do Nascimento Simões Fred Augusto Lourêdo de Brito Francisco Vanies da Silva Sá Vander Mendonça

High air temperatures and intense solar radiation in semi-arid regions may impose thermal and photooxidative challenges on cacti. This study evaluated the physiological responses of white-fleshed (Hylocereus undatus) and red-fleshed (Hylocereus costaricensis) pitaya to foliar application of calcium sources and sunscreens. The experiment used a randomized block design (4 × 2 factorial) and tested four strategies (control, SunGard®, calcium silicate, and Bordeaux mixture) on the two pitaya species. In red-fleshed pitaya, Bordeaux mixture was associated with the highest contents of chlorophyll a, chlorophyll b, total chlorophyll, and carotenoids, with increases of approximately 101.3%, 114.4%, 108.3%, and 99.4%, respectively, relative to the control, while reducing energy dissipation (DI0/RC = 0.42) and increasing the photosynthetic performance index (PIabs) by 62.0%. In contrast, white pitaya showed a marked increase in the quantum yield of dissipation (φD0) and DI0/RC (1.53 with Bordeaux mixture), indicating treatment-dependent changes in excitation-energy partitioning. Maximum photochemical efficiency (Fv/Fm) remained high in both species (approx. 0.81–0.84) under calcium silicate and SunGard® treatments. Foliar mineral formulations modified photosynthetic pigment contents and fluorescence responses in a species-dependent manner under semi-arid field conditions; the observed changes do not, by themselves, establish increased thermotolerance. Overall, the foliar formulations elicited distinct species-dependent responses in pigment accumulation, energy dissipation, and PSII photochemical performance under the recorded semi-arid field conditions.

Plants doi: 10.3390/plants15213494

Authors: Lili Guo Gongfu Shi Yao Wang Rui Xie Shuli Wei Xinhui Huang Huimin Shi Jing Fang Ying Liu Jie Ma Xiaoqing Zhao

Drought stress severely constrains flax (Linum usitatissimum L.) production, yet the transcriptional logic underlying genotypic variation in root drought tolerance remains unclear. We subjected 20 flax cultivars to drought at peak flowering, constructed a continuous Drought Tolerance Index (DTI) based on biomass maintenance, and generated 120 root transcriptome profiles. Integrative analyses combining PCA/UMAP, β–variational autoencoder (β–VAE) latent space modeling, gene co-expression-network analysis (WGCNA), Random Forest–SHAP (SHapley Additive exPlanations), and virtual knockout were performed. We found that drought-tolerant and drought-sensitive cultivars underwent transcriptional remodeling of similar magnitude, but diverged in direction along a latent strategy axis, corresponding to “water-conserving” and “growth-maintenance” strategies, respectively. The water-conserving strategy was characterized by upregulation of genes involved in stress protection, protein folding, and ABA signaling, whereas the growth-maintenance strategy showed elevated expression of oxidative and cell wall-related genes. Drought strengthened co-expression coordination within a core regulatory module—enriched for transcription factors and hormone signaling—despite its overall expression decline. A small set of leverage genes were identified that accurately predicted DTI; these genes were not network hubs, yet their virtual knockout induced substantial drift in the latent strategy space, confirming their non-redundant predictive information. This framework uncovers transcriptional strategy divergence underlying flax root drought tolerance and offers candidate targets for molecular breeding.

Plants doi: 10.3390/plants15213493

Authors: Ouli Xiao Bin Ma Xiaofeng Dai Jieyin Chen Zhiqiang Kong

Knowledge regarding the structure and distribution patterns of rhizosphere microbial communities of Astragalus mongholicus (A. mongholicus) under wild-simulated cultivation remains limited. This study investigated how cultivation duration affects soil properties, microbial communities, and bioactive compound accumulation, as well as the relationships among these factors. High-throughput sequencing of the 16S rRNA and ITS regions was performed to profile bacterial and fungal communities in rhizocompartments of 1–4-year-old A. mongholicus. Meanwhile, soil physicochemical properties and bioactive compounds were quantified. Fungal community diversity in the rhizosphere of A. mongholicus varied across cultivation years, whereas bacterial community diversity showed a comparatively weaker response. The relative abundances of the potentially beneficial taxa Mortierellomycota and Mortierella in rhizosphere soil gradually declined with increasing cultivation age. Soil nutrient-related variables (OM, OC, P, and NH4+-N) were the main drivers shaping microbial community composition. Bioactive compound contents also fluctuated across cultivation years. Microbial genera including Mortierella, Pseudomonas, Bacillus, and the Allorhizobium–Neorhizobium–Pararhizobium–Rhizobium clade may be closely associated with the accumulation of bioactive compounds in A. mongholicus. These findings suggest that cultivation duration reshaped rhizosphere communities and bioactive compound accumulation in A. mongholicus. Soil nutrient availability and beneficial microbes were closely associated with these changes, providing insights for microbiome-informed quality improvement.

Plants doi: 10.3390/plants15213492

Authors: Zihao Mao Yazhi Zhao Fen Xiao Chenyu Qi Aoqing Bian Xinyu Dou Juyang Liao Jianan Chen Liqun Huang Caixian Liu Xiaohong Wang

Hibiscus syriacus L. is a landscape plant with ornamental, economic, and ecological value. However, the low natural fruit-setting rate observed in most cultivars significantly hinders its crossbreeding, germplasm innovation, and large-scale application. We analyzed 20 cultivars to identify the key floral traits associated with H. syriacus fertility. Fifty floral phenotypic and pollen morphological indicators were systematically quantified, fruit-setting and pollen vitality rates were recorded, and insect foraging behaviors were observed to assess the diversity of floral characteristics and their correlation with fertility. The results showed that notable variations in floral phenotype and pollen morphology exist among the studied cultivars. Pollen grains were characterized by spinous ornamentation, with notable differences in grain size, spine morphology, and surface wrinkling. Correlation analysis identified nine core floral and palynological indicators that are highly associated with H. syriacus fertility. These comprise five floral phenotypic variables, such as pistil length (PL2) and heart length (HL), and four microscopic palynological features, including thorn curvature (TC) and spine number (TN). We built a phenotypic and palynological framework for analyzing fruit setting in H. syriacus. These nine key traits can act as morphological markers to quickly assess hybrid parents, supporting its genetic improvement, breeding and industrial promotion.

Plants doi: 10.3390/plants15213491

Authors: Qing Xu Dong Han Tao Li Xin Liu Jinchi Zhang

Afforestation of saline–alkali tidal flats is a key ecological engineering measure to mitigate soil degradation; however, the pathways through which different plantation types drive changes in soil multifunctionality remain unclear. We selected 15-year-old stands of 10 plantation types and a bare tidal flat (CK) in the Dafeng Forest Farm of northern Jiangsu and measured 20 soil physicochemical and biological indicators in three layers (0–20, 20–40, and 40–60 cm). Z-score standardization was used to construct carbon sequestration, nutrient cycling, enzyme activity, and comprehensive multifunctionality indices, which were analyzed via two-way ANOVA, SHAP, and PLS-PM. After afforestation, topsoil electrical conductivity decreased from 4563 to 83–202 µS cm−1 and mean weight diameter increased from 0.17 to 0.27–0.56 mm (both p < 0.001). The enzyme activity Z-score differed significantly among plantation types, and the comprehensive Z-score declined significantly with soil depth. SHAP identified R0.25, MWD, and bulk density as the dominant drivers of the comprehensive Z-score. PLS-PM (GOF = 0.615) showed that plantation types enhanced multifunctionality mainly by reducing salinity–alkalinity (path coefficient 0.96) and improving aggregate structure, whereas soil depth exerted the largest total negative effect (−0.84). U. thomasii and P. tomentosa showed preliminary potential for improving multifunctionality in this study, which requires verification across additional sites, stand ages, and seasons.

Plants doi: 10.3390/plants15213490

Authors: Fang Xu Keer Wang Yi Li Peng Zhang

In this study, dynamic headspace sampling combined with automated thermal desorption–gas chromatography/mass spectrometry (ATD–GC/MS) was employed to profile the volatile emissions from fresh flowers of 30 Chinese herbaceous peony cultivars (Paeonia lactiflora Lactiflora Group). Relative odor activity values (ROAV) were calculated to pinpoint key aroma-active constituents, and principal component analysis (PCA) was applied to examine chemometric relationships among the cultivars. The volatile blends were predominantly composed of terpenoids, benzenoids, and alcohols. Ten compounds—namely ocimene, β-terpinene, terpinolene, 1,4-dimethoxybenzene, phenethyl alcohol, 1-pentanol, eucalyptol, linalool, citronellol, and cis-3-hexenyl acetate—exhibited ROAV > 10 and were thus recognized as major odor contributors, jointly conferring rosy, citrusy, sweet, woody, and herbaceous–green nuances. Cultivar-to-cultivar variations in the dominant odorants, such as ocimene in ‘Jinzanciyu’, citronellol in ‘Chifen’, and linalool in ‘Dongfanghong’, gave rise to distinctive sensory signatures. Furthermore, PCA score plots revealed a clustering trend among cultivars with similar petal colors, suggesting a possible linkage between floral pigmentation and volatile composition. Collectively, these results offer a chemical foundation for fragrance typology and serve as a useful reference for scent-oriented breeding programs in herbaceous peonies.

Plants doi: 10.3390/plants15213489

Authors: Zhe Hao Tao Lin Shengchuan Tian Lan Du Yugang Wang Bin Zhang Xinjun Zheng

Groundwater-table decline is an important environmental driver of desert vegetation degradation in arid regions, yet population structure and growth potential of co-dominant species along groundwater-depth gradients remain insufficiently compared. To examine differences in population dynamics between Haloxylon ammodendron and H. persicum, five sites with groundwater depths of 4, 7, 10, 13, and 16 m were selected along the southern margin of the Gurbantunggut Desert. Population stage and size-class structure, seedling survival, and reproductive characteristics were surveyed, and population dynamics were evaluated using life tables, population structural dynamic indices, demographic parameters, and Leslie matrix projections. Both species had abundant seedlings but relatively few non-current-year individuals, and seedling mortality approached 98%. The density of H. ammodendron individuals in stages III–IX was relatively similar across the 4–13 m sites but substantially lower at 16 m. In contrast, the density of H. persicum was lower across the 7–16 m sites than at 4 m. With increasing groundwater depth, the intrinsic rate of increase and finite rate of increase in H. ammodendron declined significantly, whereas its population doubling time increased significantly; none of the demographic parameters of H. persicum showed a significant relationship with groundwater depth. Under constant model parameters, Leslie matrix projections indicated that H. ammodendron populations generally declined at groundwater depths of 10–16 m, whereas H. persicum populations remained relatively stable or increased. The two species showed contrasting population patterns across the groundwater-depth gradient, but the spatial design does not establish groundwater decline as their sole cause, and differences in response strength between species require direct statistical testing. The projections describe conditional trajectories rather than observed temporal changes. The findings provide a basis for population maintenance of dominant desert shrubs and vegetation conservation under continued groundwater-table decline in the Gurbantunggut Desert.

Plants doi: 10.3390/plants15213488

Authors: Chongsen Ma Xiao Liu Aimin Luo

Background: Accurate recognition of cotton growth stages in complex field environments is crucial for optimizing crop management and harvest scheduling. Methods: In this study, a lightweight multiscale heterogeneous fusion detection network, RT-HEM-Li Fusion, was constructed based on the RT-DETR framework to identify five cotton growth stages, namely cotton bud, cotton blossom, early boll, matured cotton boll, and split cotton boll. The network was evaluated on a publicly available cotton growth dataset. To enhance feature representation capability and small-object detection performance, an Inverted Residual Mobile Block was introduced into the ResNet-18 backbone, an adaptive dual-path aggregation pyramid with trident kernels was designed to preserve cross-scale details, and the baseline GIoU loss was replaced with WIoU-Focaler-NWD to enhance small-object localization and class discrimination. Results: RT-HEM-Li Fusion achieved the highest overall mAP@0.5 of 0.782 among the compared models, ahead of TridentNet (0.751) and RT-DETR (0.722) on average, while requiring only 13.905 M parameters and 40.2 GFLOPs, approximately one twentieth of the computational cost of TridentNet and fewer parameters than YOLOv5 (25.048 M), YOLOv8 (25.843 M), and DINO (47.548 M). Conclusions: RT-HEM-Li Fusion provides an effective image-based tool for monitoring cotton growth stages, demonstrating stable detection performance under the complex field conditions represented in the evaluated dataset, with a favorable balance between accuracy and model complexity.

Plants doi: 10.3390/plants15213487

Authors: Xitlalick García-Nava Miguel Fuentes-Figueroa Rosario Redonda-Martínez Laura Sepúlveda-Cuellar Joyce Trujillo Elihú Bautista

In Mexico, Asteraceae is the most diverse plant family in terms of number of species, with an occurrence of 3127 species, of which 3063 are considered native and 1996 are endemic. For this reason, the country is considered as a center of distribution and endemism. Mexican Asteraceae plants are characterized by their edible and medicinal usages for the treatment of infectious and chronic degenerative diseases, as well as for their agrochemical potential. Phytochemicals from these species have been of interest for decades and are still being considered as current sources of natural products with high structural diversity and bioactivity. The chemical constituents derived from these plants include alkaloids, flavonoids, phenolics, and terpenoids. The reported biological activities were selected according to predefined eligibility criteria. Of these, the terpenoids represent the most relevance due to the number of chemical structures described and their bioactivity as anti-inflammatory, antiprotozoal, antitumor, cytotoxic and phytotoxic agents. In this scoping review, we present the results of a bibliographic revision, and its subsequent analysis to illustrate the state of the art in natural product discovery and pharmacological prospecting with Asteraceae species native to Mexico over the last 30 years. This review highlights the chemical structural diversity of the Asteraceae family members and provides a selective overview of the agrochemical and pharmacological potential of their specialized metabolites, identifying gaps in the subject matter and trends for future research.

Plants doi: 10.3390/plants15213486

Authors: Arijit Garai Biswajit Pramanick Dhirendra K. Roy Santosh Kumar Singh Nilanjaya Vipin Kumar Md. Abbas Ahmad R. K. Ranjan

Weed management has emerged as the main constraint to the successful adoption of climate-resilient dry direct-seeded rice (DDSR) in India. Entire-crop failure has been reported in many direct-seeded rice cases, which is the main constraint to adopting this climate-resilient technology among Indian farmers. A two-year field experiment was conducted during the rainy seasons of 2023 and 2024 to identify effective herbicide combinations for the control of diverse weed flora in DDSR. The conventionally recommended herbicides such as bispyribac-sodium, pyrazosulfuron-ethyl, etc., and recently introduced herbicides like florpyrauxifen-benzyl, triafamone, etc., which have not been explored extensively in the DDSR production system in India, were evaluated. It was found that weed management practices considerably influenced the weed density, weed biomass, weed control efficiency, and grain yield in DDSR. Among the herbicide combinations evaluated, pendimethalin at 1000 g/ha as pre-emergence followed by florpyrauxifen-benzyl (1.31% w/w) at 15.3 g/ha + penoxsulam (2.1% w/w) at 20 g/ha as post-emergence at 25 days after sowing provided the most effective broad-spectrum weed control by recording the lowest overall weed density and biomass throughout the crop growth period. Whereas pendimethalin followed by bispyribac-sodium + pyrazosulfuron-ethyl was the most effective against Cyperus spp., pendimethalin followed by triafamone + ethoxysulfuron recorded the lowest density and biomass of Cynodon dactylon. Treatment T4 recorded weed control efficiencies of 88.0, 88.6, and 85.0% at 30, 60, and 90 days after sowing, respectively, and produced a grain yield of 4.67 Mg/ha, which was statistically at par with the weed-free treatment (4.80 Mg/ha). The sequential application of pendimethalin followed by florpyrauxifen-benzyl + penoxsulam provided effective broad-spectrum weed control and maintained rice productivity comparable to the weed-free treatment. Therefore, this herbicide combination is recommended for weed management in dry direct-seeded rice (DDSR) in India.

Plants doi: 10.3390/plants15213485

Authors: Giada Trebaiocchi Eleonora Spinozzi Roman Pavela Loredana Cappellacci Filippo Maggi Riccardo Petrelli

The recent outbreak of vector-borne diseases has highlighted the crucial importance of effective mosquito control programs. Nevertheless, mosquito management still represents a challenge due to the rapid onset of resistance to conventional insecticides. In this regard, there is an increasing research interest in botanicals as attracting valuable sources of insecticidal compounds. Smyrnium olusatrum L. (Apiaceae), an aromatic plant of great scientific relevance, owes its bioactivity to furanosesquiterpenes, whose major constituents include isofuranodiene (IFD), curzerene, 1-β-acetoxyfuranoeudesm-4(15)-ene, and atractylon. Since these compounds have demonstrated promising larvicidal activity against different mosquito species, in this study they were tested individually and in blends against Culex quinquefasciatus Say larvae, demonstrating the occurrence of synergistic interactions leading to enhanced larvicidal activity. In detail, the blend (1:1) consisting of curzerene and atractylon displayed the highest synergistic action (61.25% mortality), double with respect to that expected (33.75%). Moreover, the pure compounds were tested at their LC50 concentrations against the non-target species Daphnia magna Straus, Chaoborus flavicans Meigen, and Tubifex tubifex Müller, with mortalities from 7.5 to 33.7%. Overall, these findings demonstrate that synergistic furanosesquiterpene blends represent a promising botanical strategy for mosquito control, while minimizing the environmental impact associated with conventional insecticides.

Plants doi: 10.3390/plants15213484

Authors: Leila Bernart Vilela Sirlene Lopes Oliveira Amanda Ferraresi Roberto Laura Tarrafel da Silva Amanda Ribeiro Peres Portugal Letusa Momesso Carlos Alexandre Costa Crusciol

Maize production in Brazil is concentrated in the off-season, after soybean harvest. Reduced water availability can compromise maize growth and productivity. Foliar fertilization may improve crop development and yield, although evidence on simultaneous application of macronutrients remains limited. This study evaluated the effects of foliar nitrogen–phosphorus–potassium (NPK) formulations application on off-season maize physiology and grain yield. We compared foliar NPK application with an untreated control over two growing seasons. Leaf nutritional status, photosynthetic enzymes (ribulose-1,5-bisphosphate carboxylase/oxygenase [Rubisco] and phosphoenolpyruvate carboxylase [PEPcase]), gas exchange, oxidative stress indicators (H2O2 and MDA), antioxidant enzymes (superoxide dismutase [SOD] and catalase [CAT]), protein and sugar concentrations, yield components, and grain yield were evaluated. Foliar NPK application improved maize nutritional status and photosynthetic performance Rubisco activity, gas exchange, water use efficiency (WUE) in 2020, and protein and sugar concentrations. Foliar NPK reduced hydrogen peroxide (H2O2) accumulation and lipid peroxidation, measured as malondialdehyde (MDA) in both seasons, accompanied by lower antioxidant enzyme activities, suggesting reduced oxidative pressure and antioxidant demand. Grain yield increased by 0.72 Mg ha−1 (11.5%) and 0.64 Mg ha−1 (12.0%) in the first and second seasons, respectively, compared with untreated maize. NPK improved physiological performance and grain yield of off-season maize, supporting its use as a complementary practice.

Plants doi: 10.3390/plants15213483

Authors: Adam R. Herdman Ernesto B. Mújica Lawrence W. Zettler Elaine H. González Alejandro Camejo Vergara Mark W. Danaher Michael E. Kane Héctor E. Pérez Carrie Reinhardt Adams

The ghost orchid, Dendrophylax lindenii, is a rare leafless epiphyte restricted primarily to southern Florida and western Cuba. Despite a decade of conservation efforts, the understanding of in situ long-term stressors remain understudied. Building on previous demographic studies, we extended annual monitoring of D. lindenii at the Florida Panther National Wildlife Refuge from 2015 to 2024 and at Guanahacabibes National Park, Cuba, from 2006 to 2023, to provide a direct long-term comparison of population trajectories across two major portions of the species range. We evaluated total trends and regional differences, changes in demographic size structure, and associations between annual population growth and regional climate conditions and disturbance. Both populations declined through time, but the decline was greater in Florida, where totals decreased by 8.8% every year from a maximum of 529 individuals in 2017 to 237 in 2024, compared with a decline of 2.5% from 360 individuals in 2006 to 195 in 2023 in Cuba. Plants were grouped by mean living root length C1 (0–8.0 cm), C2 (8.1–11.3 cm), C3 (11.31–15.3 cm), and C4 (>15.3 cm). In Florida, C1 and C3 declined significantly, while losses of large C4 plants also occurred. In Cuba, C1, C2, and C3 declined while C4 increased, retaining a greater number of large reproducing plants. Flowering fluctuated in Florida but increased in Cuba, while fruit production remained low in both regions. Regional temperatures increased during the study period, but annual population size was not statistically associated with temperature, precipitation, or tropical cyclone occurrence. Class-based projections indicated continued declines in both regions, with a stronger projected decrease in Florida. These declines indicate that conservation actions should be tailored to local population structure. Protection of existing large reproductive plants, supplementation of depleted smaller-size classes, and coordinated management across remaining populations may provide the strongest basis for sustaining D. lindenii in Florida and Cuba.

Plants doi: 10.3390/plants15213482

Authors: Juthaporn Saengprajak Pimradawan Kulchotchaiwong Aphidech Sangdee

Aromatic rice is highly valued worldwide for its distinctive fragrance and superior grain quality. Despite the importance of genetic diversity and aroma-related metabolites for germplasm conservation and breeding, studies integrating molecular markers and volatile profiling in Thai rice remain limited. This study integrated ISSR, SCoT, and SRAP molecular markers with static headspace gas chromatography-mass spectrometry (HS-GC-MS) to characterize the genetic diversity and volatile profiles of Thai rice germplasm. Twenty-nine aromatic and non-aromatic cultivars were evaluated using the three molecular marker systems. All marker systems revealed substantial polymorphism, with SRAP showing the highest overall discriminatory performance, including the highest percentage of polymorphic bands (79.19%) and the highest mean PIC, RP, MI, and He. The combined marker dataset provided a broader representation of molecular variation, with UPGMA clustering and PCoA differentiating the rice accessions into distinct genetic groups. Four representative cultivars (KDML105, PTT1, RD43, and RD79) were subsequently selected from the 29 rice cultivars for comparative volatile profiling based on their genetic relationships. Six volatile compounds were tentatively identified, comprising four aldehydes, one carboxylic acid, and one ester. Hexanal was the predominant volatile compound across all four cultivars, whereas acetic acid, 4-methylbenzaldehyde, and methyl salicylate differed among the analyzed samples. Multivariate analysis distinguished KDML105 from the other three cultivars based on their volatile profiles. Integrating multilocus DNA fingerprinting with volatile profiling provides complementary insights into the genetic and biochemical diversity of Thai rice germplasm, supporting cultivar characterization, germplasm management, and rice breeding.

Plants doi: 10.3390/plants15213481

Authors: Claudia Cruz Paulina Villamahua Martín Urgilés-Recalde Omar Malagón Vladimir Morocho Yunierkis Perez-Castillo

The leaf essential oil of Croton chimboracensis, a species native to Ecuador, was investigated using an integrated chemical, biological, and computational approach. Volatile constituents were identified by gas chromatography–mass spectrometry (GC–MS), and their relative proportions were determined by gas chromatography–flame ionization detection (GC–FID) peak-area normalization. Forty-seven compounds were identified, accounting for 95.17% of the total GC–FID peak area. The oil was dominated by hydrocarbon sesquiterpenes (72.75%), with trans-muurola-4(14),5-diene (34.06%), longifolene (15.34%), bicyclogermacrene (7.65%), caryophyllene oxide (5.53%), α-humulene (4.38%), and γ-cadinene (4.02%) as the principal constituents. The essential oil exhibited an assay-dependent radical-scavenging response, with an SC50 value of 15.20 µg/mL in the 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay (95% confidence interval CI: 8.97–19.83 µg/mL) and 2983 µg/mL in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay (95% CI: 2719–3378 µg/mL). The essential oil also inhibited acetylcholinesterase (AChE), with an IC50 value of 43.78 µg/mL (95% confidence interval: 40.68–47.05 µg/mL). Molecular docking and absolute binding free-energy calculations were performed for all 47 identified constituents. These calculations prioritized spirolepechinene, β-phellandrene, β-pinene, alloaromadendrene epoxide, and (2E,4E)-nona-2,4-dien-1-ol as potential AChE-binding constituents, with predicted binding free energies of −20.22 ± 0.93, −16.59 ± 0.87, −16.33 ± 0.91, −15.81 ± 2.53, and −15.29 ± 1.91 kcal/mol, respectively (ΔG ± SEM). These findings identify candidates for subsequent experimental validation and provide molecular hypotheses concerning their possible contributions to the AChE-inhibitory activity of the complete oil. Overall, this study provides an integrated characterization of C. chimboracensis essential oil and establishes a basis for further investigation of its antioxidant and acetylcholinesterase-inhibitory constituents.

Plants doi: 10.3390/plants15213480

Authors: María Cecilia Castro Alejandro Diaz Guillermo Schinella Flavio Francini

Background: Obesity, type 2 diabetes mellitus (T2DM) and cardiovascular diseases (CVDs) are growing public health problems that affect an increasing number of people worldwide because of their prevalence, costs, and health effects. Medicinal plants have preventive effects against chronic diseases, and the trend towards seeking natural products for their treatment and prevention is growing. Yerba mate (YM) is an herbal tea prepared from the dried and roasted leaves of a native South American tree, Ilex paraguariensis St. Hil. It is known for its high content of polyphenols, methylxanthines, and saponins. Methods: Searches were conducted in PubMed/MEDLINE, Scopus, and the Cochrane Central Register of Controlled Trials; Google Scholar was used as a supplementary source for citation tracking and identification of potentially relevant records. For clinical trials, articles published between 2011 and 2026 were selected. Results: Several studies found improvements in serum parameters, weight gain and reduced inflammation and oxidative stress in animal models. In humans, few clinical trials have been performed. They suggest positive effects on human health, decreasing waist-to-hip ratio, BMI, and improving fasting glucose, HbA1c, lipid and antioxidant serum levels, and reducing the risk of developing coronary disease. Conclusions: This review integrates preclinical and clinical evidence on Ilex paraguariensis in obesity, T2DM, and CVD, with particular emphasis on the common roles of oxidative stress, inflammation, insulin resistance, and lipid dysregulation and the limitations that currently hinder clinical translation. Although YM represents a promising source of bioactive compounds and a potentially useful functional beverage, current human evidence remains insufficient to establish it as a routine treatment or preventive intervention for these cardiometabolic entities. Standardized preparations, adequately powered randomized trials, clinically relevant endpoints, and systematic long-term safety assessment are needed. By bringing these outcomes together, the review aims to distinguish mechanistic evidence from preliminary human findings and to identify priorities for standardized preparations and adequately powered clinical trials.

Plants doi: 10.3390/plants15213478

Authors: Jue Zhao Shuo Zhang Si He Qinxu Song Nicheng Wu Hucheng Xing

Agronomic regulation is vital for improving the phytoextraction efficiency of economic crops in cadmium (Cd)-contaminated soils. Here, a two-season pot experiment evaluated the effects of foliar versus soil KH2PO4 applications on ramie (Boehmeria nivea L.) under Cd stress. Growth and phytoextraction performance peaked during the second harvest season. Foliar application of 0.2% KH2PO4 increased ramet number by 118.2% and alleviated oxidative damage by sequentially activating SOD, POD, and CAT. Subcellular analysis indicated that foliar KH2PO4 enhanced Cd sequestration within root cell walls, restricting excessive Cd translocation to photosynthetic tissues. In contrast, high-dose soil application exacerbated toxicity and inhibited biomass accumulation. Structural equation modeling showed that shoot biomass primarily drove total Cd extraction (β = 0.925), whereas stem cell wall Cd retention served as the major limiting factor (β = −0.471). Overall, low-dose foliar KH2PO4 provides a promising agronomic approach to enhance Cd phytoextraction by maximizing shoot biomass while limiting tissue toxicity.

Plants doi: 10.3390/plants15213479

Authors: Marijenka Tabakovic Vojka Babic Vesna Peric Vesna Dragicevic Anika Kovincic Violeta Oro Rade Stanisavljevic Ivana Zivkovic Violeta Mickovski Stefanovic Natalija Kravic

During the transition from seed quiescence to active seedling development—where early metabolic repair and structural biomass deposition are tightly regulated by genotypic identity—hydropriming efficacy exhibits a pronounced temperature- and immersion duration-dependent trade-off. This study evaluates the capacity of hydropriming to regulate this morpho-physiological and metabolic pattern in investigated maize landraces by contrasting the unprimed baseline (T0) against treatments at 20 °C (T1–T3) and 30 °C (T4–T6) across 8-h, 16-h, and 24-h immersion periods. Under optimal eustress (during the 24-h immersion at 20 °C under T3), a high germination capacity of 94% expressed by genotype L3 relies upon active phytate phosphorus (Pphy) breakdown and robust glutathione buffering to increase chlorophyll b up to ~96%, whereas thermal strain (induced by 8-h to 24-h exposures under T4–T6) shifts metabolic priority downward into root-zone defences at the expense of chlorophyll accumulation (which declines up to ~88%). Under thermal strain, Pphy retention triggers a massive compensatory upregulation of redox status, expanding root glutathione synthesis up to ~282% and stimulating secondary phenylpropanoid pathways to drive root yellow pigment accumulation up to ~887%, significantly reinforcing integrated radical scavenging activity to mitigate oxidative damage. Principal component analysis resolves these responses into parallel, tissue-specific functional domains: optimal growth, thermal adjustment, metabolic latency, and suppressed vigour. Ultimately, the artificial neural network modelling establishes genetic identity as the predominant driver of overall system variance, verifying that the 24-h immersion at 20 °C triggers the physiological threshold required to maintain chlorophyll homeostasis for successful autotrophic establishment, providing a targeted pre-sowing protocol for routine maize germplasm regeneration.

Plants doi: 10.3390/plants15213476

Authors: Shahzad Hussain Shah Kristi Spieleder Ryan Koeth Changbin Chen

Silicon deposition in grasses produces silica bodies with characteristic morphologies and distributions, but our understanding of their organ-specific diversity and developmental silicification remains limited beyond extensively studied crop species. Here, we investigated silica body formation in Zingeria biebersteiniana, a diploid grass with 2n = 4 chromosomes, by combining organ-level morphological characterization with a post-transplant developmental analysis. Dry-ash preparations were examined by fluorescence microscopy and quantified using digital image analysis. Mature soil-grown plants exhibited organ-associated silica body morphotypes in leaf blades, leaf sheaths, stems, glumes, lemmas, and paleae, with marked differences in morphology, distribution, and epidermal coverage. Plants maintained in vitro on Murashige and Skoog (MS) medium without intentionally added Si showed little to no detectable leaf silicification. Following transfer to soil, silica bodies were readily detectable by Day 7 in both Leaf 3, which was fully expanded at transplantation, and the younger, developing Leaf 2. Descriptive analysis showed broad silica-body area distributions that shifted during the post-transplant period. Spatial imaging further revealed interplant heterogeneity: three plants showed more distal and marginal silicification, whereas nine showed relatively uniform blade-wide patterns. These results provide an anatomical and developmental framework for investigating silicon biology in Z. biebersteiniana and demonstrate the utility and limitations of fluorescence-based dry-ash imaging for descriptive analysis of silicification in grasses.

Plants doi: 10.3390/plants15213477

Authors: Yinli Shan Nan Zheng Yupeng He Enguang Nie Haiyan Wang

Pharmaceutical and Personal Care Products (PPCPs) are emerging aquatic contaminants. Existing studies investigating the uptake of PPCPs by aquatic plants under hydroponic conditions have mostly been performed in non-competitive systems. Few reports are available regarding plant uptake and translocation of PPCPs under multi-species coexistence and interspecific competition. In this study, competitive hydroponic experiments were conducted in a greenhouse using 14C-triclosan to explore the uptake and distribution of triclosan in six hydrophytes: water spinach, purple perilla, cress, penny grass, cane shoot, and rice. The results showed that the triclosan uptake capacity of aquatic plants in the competitive hydroponic system followed the order: cress > water spinach > cane shoot > rice > penny grass > purple perilla. The root 14C-triclosan concentrations in water spinach (222.15–564.54 μg g−1) and cress (198.66–569.25 μg g−1) were significantly higher than those of the other species. Triclosan mainly accumulated in roots (90.36–99.04% of total plant uptake), with shoot levels 2–3 orders of magnitude below roots. Compared with non-competitive hydroponic conditions, interspecific competition exerted no significant effects on the uptake, distribution and translocation of triclosan in plants. This work advances the understanding of triclosan accumulation-translocation in hydrophytes under competitive conditions and supports risk assessment for triclosan.

Plants doi: 10.3390/plants15213475

Authors: Jianzhong Sang Xiaofang Ding Jiquan Zhang Bingyan Liu Xiaoming Lu Zhuanhong Liang Tingting Fan Lijun Liu Yuanyuan Pu Junyan Wu Li Ma Wangtian Wang Gang Yang Wancang Sun

Winter turnip rape (Brassica rapa L.) is an important winter oilseed crop in the arid and cold regions of northern China due to its strong cold tolerance and wide adaptability. However, its prevalent self-incompatibility readily causes inbreeding depression, leading to declines in yield and quality and creating difficulties for cultivar breeding and pure-line propagation. To clarify how self-compatibility affects growth, yield, quality, and physiological traits, highly self-compatible (HSC), self-compatible (SC), and self-incompatible (SI) lines were compared under self-pollination and open pollination, and leaf antioxidant enzyme activities and endogenous hormone contents were measured. The results showed that the effects of selfing on seedling vegetative growth, agronomic traits, quality, and biomass were far smaller in HSC and SC lines than in SI lines, with inbreeding depression markedly alleviated. The mean inbreeding depression rates (IDRs) of major traits in HSC, SC, and SI lines were 4.32%, 8.14%, and 20.05%, respectively, the IDRs of effective silique number per plant were 3.03%, 5.22%, and 20.53%, and those of seed yield were 4.69%, 9.39%, and 27.90%. Line-level ANOVA and Tukey tests (three lines per group) confirmed that the IDRs of SI lines were significantly higher than those of HSC lines for all six core traits, with SC lines intermediate. Physiological and biochemical analyses indicated that HSC lines maintained higher SOD activity, and the contents of growth-promoting hormones, such as gibberellin (GA), cytokinin (CTK), and auxin (IAA), were generally higher than in SI lines, whereas CAT activity did not show a consistent advantage: in HSC lines CAT did not differ significantly between SP and OP (the two shared a significance group), and under open pollination SI-OP was grouped with HSC-OP and SC-OP. In contrast, after selfing, SI lines accumulated more abscisic acid (ABA), whereas the ethylene (ETH) differences were not consistently significant and the coordination of their antioxidant enzymes was altered rather than disrupted. Correlation analysis showed that the contents of growth-promoting hormones (GA, CTK, IAA, and BR) were significantly or highly significantly positively correlated with yield traits, such as seed yield per plant, effective silique number per plant, and thousand-seed weight, whereas ABA and ETH contents were highly significantly negatively correlated. A structural equation model, based on composite scores of hormones, enzymes, growth, and yield and quality, indicated that the hormone composite was strongly associated with the growth composite (standardized path coefficients of hormone → growth and growth → yield and quality were 0.816 and 0.535, respectively), whereas the enzyme composite was directly associated with yield and quality (0.291). These paths reflect statistical associations among the composite scores rather than confirming the specific underlying mechanisms, and vegetative growth was the main mediator linking the hormone composite to yield and quality. To our knowledge, self-compatibility in winter turnip rape can maintain relatively stable plant growth and development as well as yield and quality by alleviating inbreeding depression and maintaining physiological metabolism and hormone homeostasis. This study provides a theoretical basis for the screening and utilization of self-compatible germplasm and for high-yield breeding of winter turnip rape; because stress tolerance was not directly measured, any extrapolation to stress-resistant breeding requires dedicated stress-tolerance experiments in future work.

Plants doi: 10.3390/plants15213474

Authors: Guanglong Zhu Chenghao Hu Qingqing Geng Qiwei Yang Sizhuo Chen Yuewen Ding Hussam Ahmad Tiankang Zhu Wen Sun Irshad Ahmad Guisheng Zhou

Arable land is diminishing rapidly as a result of environmental degradation and urbanization. The development and utilization of saline–alkali soils therefore represent an important strategy for ensuring global food security. Rice (Oryza sativa L.) is a staple food for over half of the global population, yet its productivity is severely constrained by saline–alkali stress. Salinity (mainly NaCl) and alkalinity (high pH caused by Na2CO3/NaHCO3) markedly impair both yield formation and grain quality. This review establishes diagnostic thresholds for salinity (ECe) and alkalinity (pH) to mitigate their adverse effects. The effects of saline–alkali stress on rice germination, seedling establishment, plant growth, reproductive development, and grain filling are summarized. And the underlying physiological and molecular mechanisms are elucidated, including ion homeostasis (mediated by OsHKT1;5, OsSOS1, and OsNHX1), osmotic adjustment, as well as alkalinity-specific processes such as rhizosphere acidification, organic acid exudation, and Fe/Zn acquisition under high pH, endogenous hormone crosstalk, antioxidant defense, starch/protein biosynthesis, and related key genes and quantitative trait loci (QTLs). Furthermore, recent advances of stress-resistant cultivation and regulation strategies are highlighted. On this basis, future research directions are prospected. This review provides a comprehensive understanding of the physiological and molecular mechanisms underlying rice saline–alkali tolerance, with important implications for achieving high yield and quality in rice production on saline–alkali soil.

Plants doi: 10.3390/plants15213473

Authors: Md Minarul Islam Yunseon Hwang Yongsoo Choi Ki Sung Kang

The rhizosphere microbiome plays a crucial role in the development, health, and therapeutic properties of Panax ginseng. Therefore, we evaluated the spatial trends in bacterial communities using a comprehensive reanalysis of 359 sequenced rhizosphere samples from seven NCBI Sequence Read Archive BioProjects. The SILVA reference database was used to compare the Chinese and Korean sequencing datasets, focusing on quality filtering, denoising, amplicon sequence variants, and taxonomy assignment. The bacterial community structure was characterized, and differentially abundant taxa were identified using the Kruskal–Wallis test with false discovery rate correction. In addition, based on the detected bacterial taxa, potential ecological traits were contextually interpreted to provide insight into their potential ecological significance. Despite regional and technical variations, 37 bacterial taxa consistently emerged as conserved core microbiome members. Regional analyses revealed distinct bacterial drivers, such as Ligilactobacillus, Romboutsia, Nitrospira, and Bradyrhizobium in the Korean samples and Holophaga and Arachidicoccus in the Chinese samples. Putative functional annotations showed that plant-associated defense mechanisms, nutrient mobilization, and carbon cycling varied geographically. Overall, these findings indicate that although a conserved core community exists, regional differences may influence the composition, functional potential, and ecological relationships of the P. ginseng rhizosphere.

Plants doi: 10.3390/plants15213472

Authors: Min-Kyu Park Jong-Du Lee Gun-Myung Lee Yeong-Jun Park Xiaofei Dang Yuxin He Hyuk-Joon Kwon Jae-Ho Shin

Wild-simulated and forest-grown Panax ginseng commands substantially higher prices than field-cultivated ginseng, largely on the assumption of a superior ginsenoside profile. Following PRISMA-ScR, we searched PubMed and Crossref (14 August 2026) and hand-searched Korean-language journals; 270 of 300 records were retained, and 4199 ginsenoside values were extracted from 44 studies, with the study as the unit of synthesis. Of 18 within-study contrasts in P. ginseng, eight reported higher major-ginsenoside abundances in forest material, but 13 of the 18 contrasts were not age-matched. The five age-matched comparisons were mixed: higher relative abundance of 19 constituents in one untargeted study, higher Re only in another, higher totals but lower Rg1 and Rf in a third, and rare-ginsenoside differences in two, with major ginsenosides either not consistently different or not reported. Truly wild ginseng is represented by fewer than ten quantified roots. Three conclusions follow: (i) forest-grown roots are chemically distinguishable, but consistent enrichment of major ginsenosides has not been shown once age is considered; (ii) largely uncontrolled age, site, plant part, and germplasm produce variation comparable to the reported system differences; and (iii) age-matched, germplasm-controlled studies with harmonized reporting are needed before profile-based quality claims are justified.

Plants doi: 10.3390/plants15213471

Authors: Alessandro Borrelli Farzaneh Zamani Marco Esposito Roberta Paradiso

According to the list of Traditional Agri-food Products (TAPs) of the Italian Ministry of Agriculture, Food Sovereignty and Forestry, 1636 plant-based TAPs are currently registered in Italy. This study aimed at identifying strengths and constraints associated with the production, commercialization, and conservation status of plant-based TAPs selected in Campania. Data on all 246 TAPs recorded in the region were collected and analysed to assess their distribution in the 5 provinces. The conservation status was assessed by cross-referencing the TAPs list with the Regional Register of Plant Genetic Resources at Risk of Extinction and the National Register of Conservation Varieties. The study was conducted on a sample of 13 products, selected by edible organ (leaf, fruit, inflorescence, seed, and tuber/rhizome). Questionnaires concerning production and commercialization were distributed to two farmers per product. Most farms showed limited surfaces (<1 ha in 69.2%), with low yearly production levels (<10 q in 27% and 10–50 q in 38%). Cultivation mainly consists of organic farming (58%), reflecting the widespread adoption of low-input and environmentally sustainable practices, but entails higher production costs and operational constraints, affecting farm economic sustainability and competitiveness in broader markets. All farms distribute products at the local level, while 15% and 23% also reach national and European markets, respectively. Our results highlight the potential role of TAPs in supporting the sustainable utilisation of local plant genetic resources, territorial identity and rural food tourism. Strengthening the economic viability of plant-based TAPs may contribute to reducing the risk of genetic erosion and preserving Campania’s plant biodiversity and cultural heritage.

Plants doi: 10.3390/plants15213470

Authors: Chang’An Zhou Chenxiao Wang Xiumin Guan Shanxing Zhao Jiarui Yang Laichen Zheng Jing Zhang Xiaoying Ning Keyan Peng Jiutao Yang Yuan Yao Lanping Luo Xin Liu

Photosynthetic source strength is a major determinant of soybean yield formation through its regulation of reproductive sink development. However, high-throughput quantification of sink traits under contrasting photosynthetic source conditions remains a major bottleneck for field-based source–sink physiology research. In this study, three soybean cultivars were subjected to graded artificial defoliation (0%, 25%, and 50%) at the R2 flowering stage to establish different levels of defoliation-induced source limitation. Biological observations showed that mature pod number consistently declined with increasing defoliation intensity across all cultivars, supporting the critical role of flowering-stage source limitation in determining sink formation. To enable efficient quantification of these sink responses, Unmanned Aerial Vehicle (UAV)videos acquired at maturity were used to construct a field soybean pod dataset, and an integrated detection–tracking framework, PodCountNet, was developed by combining an improved SPDM-YOLO detector with the OC-SORT tracking algorithm. The proposed SPDM-YOLO achieved a precision of 88.2% and an mAP@0.5 of 94.5% under diverse canopy conditions. Although single-view counting accuracy varied with canopy architecture and illumination, dual-view fusion effectively reduced systematic errors and, achieving MAE = 66.3 pods and RMSE = 88.0 pods at the individual plot level (n = 27), substantially outperforming single-view front-lit (MAE = 114.2) and back-lit counting (MAE = 172.9). The proposed framework enables accurate, non-destructive, and high-throughput quantification of soybean sink responses under contrasting photosynthetic source conditions, providing an effective phenotyping tool for investigating source–sink coordination and supporting photosynthesis-oriented soybean breeding.

Plants doi: 10.3390/plants15213469

Authors: Wei Chu Hezhi Lin Jianjuan Li Miao Jia Shenghang Wang Mingzhe Li Sansan Liu Zeyan Wu Wenxiong Lin

Plant growth-promoting bacteria can influence host metabolism, but live-inoculation systems confound chemical signaling with colonization, nutrient exchange, and continuous metabolite release. Here, Paenibacillus cellulositrophicus LB46 was co-cultured with sterile Casuarina equisetifolia seedlings for 5 d. After 0.22 μm filtration, the interaction-conditioned filtrate was applied to new sterile seedlings; no culturable LB46 was detected in the filtrate under the plating conditions used. Untargeted LC–QTOF-MS profiled roots after 3 and 5 d and shoots after 5 d. The filtrate induced broad metabolic remodeling, with stronger responses in roots than in shoots. A total of 473, 437, and 125 metabolites met the differential-abundance criterion (BH-FDR < 0.05) in 3 d roots, 5 d roots, and 5 d shoots, respectively; 273 root metabolites showed a significant treatment × time interaction (BH-FDR < 0.05). Temporal classification revealed pronounced turnover among early-transient, late-induced, and sustained responses. At 5 d, only 11 metabolites met the differential-abundance criterion in both roots and shoots, indicating predominantly tissue-differentiated remodeling. These results define temporally and spatially differentiated metabolic responses to the interaction-conditioned environment. Because bulk physicochemical properties of the filtrates were not characterized, the observed responses cannot be attributed exclusively to specific chemical signals.

Plants doi: 10.3390/plants15213466

Authors: Tian Gao Lei He Yuan Wang Shaofeng Su

Astragaloside IV is the best-characterized cycloartane-type triterpenoid saponin and the pharmacopoeial quality marker of Astragali Radix, with documented cardioprotective, antitumor, and immunomodulatory effects. Supply is constrained by long cultivation cycles that vary with production practice, replant disease, and phytochemical variability. Six representative genome assemblies are summarized for the genus, and they show that triterpenoid tailoring genes are partly, but not universally, clustered: a ~4 Mb biosynthetic gene cluster houses the oxidative and glycosylating enzymes, whereas the backbone synthase gene AmOSC3 lies outside it. The core route from 2,3-oxidosqualene to astragaloside IV has been reconstituted enzyme by enzyme; the in planta downstream routing, the acetylation steps forming astragalosides I and II, and the regulation, compartmentalization, transport, and storage of these saponins remain unresolved. Heterologous production is at the proof-of-concept stage: 1.04 mg/L cycloastragenol in yeast (versus the 25 g/L artemisinic acid precedent), 2.224 mg astragaloside IV/g dry weight in Nicotiana benthamiana, and a 4.2-fold total-astragaloside increase in AmUGT15-overexpressing hairy roots. This review consolidates genome mining, pathway biochemistry, and synthetic-biology engineering; benchmarks current platforms against pharmacopoeial quality requirements; and maps the bottlenecks between proof of concept and sustainable biomanufacturing.

Plants doi: 10.3390/plants15213468

Authors: Luis Francisco Fernández-Pozo María Angeles Rodríguez-González

Understanding how management and seasonality are associated with the soil microbiome is relevant for the sustainability of Mediterranean viticulture. This observational study compared the rhizosphere microbiome of rainfed and irrigated ‘Pardina’ vineyards on Calcaric Cambisols (Tierra de Barros, Extremadura, Spain) in five sampling campaigns (spring 2024–spring 2026) using 16S rRNA gene and ITS1 amplicon metabarcoding. The 30 soil samples were aggregated into ten community profiles per marker (five campaigns × two management), so inference is exploratory and limited by the absence of plot-level replication. Fungal communities in irrigated vineyards showed lower observed genus richness (median 180 vs. 297), Shannon diversity (3.05 vs. 3.84) and evenness than rainfed ones, with the same direction in all five campaigns, and a tendency toward higher dominance of a few genera (Berger–Parker 19.3% vs. 11.9%; not significant after correction), whereas prokaryotic richness and Shannon diversity did not differ consistently. Bray–Curtis PERMANOVA indicated a modest association between management and prokaryotic composition (R2 = 0.237, p = 0.048), which was not robust to permutations restricted within campaigns, and no detectable association for fungi (R2 = 0.151, p = 0.269); both results were sensitive to the inclusion of the spring 2026 campaign. The season and its interaction with management were not significant. Soil pH, available phosphorus and organic carbon differed between managements and planting density differed by design, so the patterns cannot be attributed to irrigation itself. The bacteria-to-fungi ratio, a sequencing-derived index, is reported descriptively only. These results identify candidate taxa and hypotheses to be tested in replicated experiments that include soil moisture and functional measurements.

Plants doi: 10.3390/plants15213465

Authors: Jing Guo Jingyi Xu Yin Zhu Qiang Cai

Cistanche deserticola is a holoparasitic medicinal plant that has lost the ability to photosynthesize and grows entirely underground, so its reduced genome might suggest weak tissue-level metabolic partitioning. Using untargeted metabolomics of 132 samples from 22 plants, with each stem divided by tissue type (outer fraction vs. internal tissues) and longitudinal position (upper, middle, lower), we found a clear two-axis organization. Tissue type accounted for the smallest proportion of variance (15.2%) yet yielded the clearest discrimination: phenylethanoid glycosides (PhGs) and flavonoids were enriched in the outer fraction, whereas iridoids and primary metabolites predominated in internal tissues; along the stem, the upper part was an active metabolic zone and the lower part a sugar sink. The decline in PhG abundance with plant size occurred predominantly within tissues, with anatomical dilution contributing only 18.6%. Pathway-resolved mapping showed that the downstream steps of PhG assembly (acylation, rhamnosylation, hydroxylation, glucosylation) are progressively enriched in the outer fraction, so the discarded outer fraction is consistent with a possible site of biosynthesis, rather than merely a storage site, for acteoside and echinacoside. This stable organization likely reflects developmental canalization, indicating that gene loss does not eliminate tissue-level metabolic specialization.

Plants doi: 10.3390/plants15213467

Authors: Qi Xu Changkun Ma Quanjiu Wang Qiuping Fu

How nitrogen (N) supply should be matched to increased planting density in drip-fertigated maize in arid regions remains unclear. A two-year split-plot field experiment (2024–2025) at Shihezi, northern Xinjiang, China, tested two planting densities (D1, 60,000; D2, 90,000 plants ha−1) and two N rates (N1, 180; N2, 240 kg N ha−1) in film-mulched, drip-irrigated spring maize. D2 increased grain yield by 13.9% over D1, because a 46.8% increase in ear number outweighed decreases in kernels per ear and kernel weight. The yield response to N depended on density (D × N, p < 0.01): N2 increased yield by 13.8% under D2 but by only 3.6% (not significant) under D1. N2 increased N accumulation at silking similarly at both densities, whereas its post-silking effects were much larger under D2, where it reduced the post-silking decline in leaf area index from 67.3% to 51.2% and increased post-silking dry matter accumulation and apparent N uptake by 22.0% and 44.3%. D2N2 produced the highest yield (11.51 t ha−1), whereas D2N1 achieved the highest partial factor productivity of N (56.2 kg kg−1). At this single site, dense planting benefited more from the higher N rate, most evidently after silking; validation across sites, hybrids, and N rates is needed.

Plants doi: 10.3390/plants15213464

Authors: Zilin Wu Xueting Li Rui Liu Qiaoying Zeng Fangyin Pan Nannan Zhang Jiayun Wu Qinnan Wang

Sugarcane is an essential global industrial cash crop for sugar and bioenergy production, and breeding superior varieties is critical to improving industrial benefits and adapting to changing climatic environments. This review synthesizes the published breeding history, agronomic trial records, juice-quality evaluations, and transcriptomic analyses of the elite sugarcane variety YT93-159, and uses this variety as a model case to discuss the achievements and challenges of sugarcane breeding in South China. YT93-159 was bred through artificial hybridization using YN73-204 as the female parent and CP72-1210 as the male parent. Data compiled from variety-registration archives, peer-reviewed studies, and the authors’ previously published transcriptomic analyses indicate that YT93-159 integrates excellent parental advantages, with prominent early sugar accumulation, high yield, stable stress resistance, and wide adaptability. On the basis of differentially expressed genes identified in published transcriptomic datasets, its superior comprehensive traits may be co-regulated by candidate pathways including carbon fixation, sucrose metabolism, and plant hormone signal transduction; however, several pathway members showed the opposite expression changes, and these mechanisms should be regarded as hypotheses requiring further functional validation. This variety has achieved large-scale popularization in China and possesses excellent functional nutritional values. By clarifying the phenotypic and proposed molecular basis of YT93-159′s superior traits, this review provides a valuable germplasm resource profile and theoretical reference for sugarcane genetic improvement and sustainable industrial development.

Plants doi: 10.3390/plants15213463

Authors: Zhihuan Ge Mengyi Yu Yinge Xie Haoran Wang Shaojun Ma Yijian Liu Shanshan Zhou Xueliang Sun Feifei Yu Mingxia Wang

The purple leaves of Brassica juncea are abundant in anthocyanins that confer stress tolerance, nutritional benefits, and ornamental value. In this study, we constructed F2 and BC1 segregating populations by crossing the purple leaf inbred line (ZY) with the green leaf inbred line (LY) to investigate the genetic basis of leaf pigmentation. Bulked segregant analysis sequencing (BSA-Seq) and map-based cloning localized the BjZY locus to a 162.08 kb interval on chromosome BB_Chr05. Based on sequence alignment and homology analysis, an R2R3 MYB transcription factor, BjMYB113 (BjuVB05G51160), was identified as a strong candidate gene, with 3 SNPs and 25 bp insert variants in the promoter region. The expression of BjMYB113 in purple leaves was significantly higher than that in green leaves, leading to anthocyanin biosynthesis and accumulation. The transient expression of BjMYB113 in tobacco leaf confirmed that BjMYB113 positively regulates anthocyanin biosynthesis. RNA-seq analysis revealed that BjMYB113 was involved in flavonoid and anthocyanin biosynthesis pathways and regulated the expression of anthocyanin-related genes, such as BjCHS, BjDFR, and BjANS. Yeast one-hybrid assays showed that BjMYB113 activated the expression of BjDFR by directly binding to its promoter. These results provide further understanding of the genetics of purple leaf and potential applications for marker-assisted breeding in B. juncea.

Plants doi: 10.3390/plants15213462

Authors: Stefanos K. Soultatos Maria Megariti Nikolaos E. Krasagakis George Papadakis Stella Chatziioannidou Despoina Kourougkiaouri Electra Gizeli Emmanouil A. Markakis

In the present study, a novel method was developed and validated for the early, rapid and on-site detection of Botrytis cinerea in crude tomato samples using a loop-mediated isothermal amplification (LAMP) assay adapted for the portable device PEBBLE. Previously reported LAMP primers (targeting the IGS region of rDNA gene) were utilized and optimized in terms of concentration for this workflow, and the sensitivity, specificity and accuracy of the developed method were determined (97.96%, 100% and 98.70%, respectively). The LAMP assay, combined with the portable device, enabled real-time quantitative colorimetric (qcLAMP) detection of B. cinerea in less than 30 min. The limit of detection (LoD) of the qcLAMP assay was 1 pg genomic DNA (≈20 genome copies) per reaction, whereas conventional PCR (utilized as the gold standard method) detected no less than 100 pg of genomic DNA per reaction. In terms of B. cinerea infective propagules, qcLAMP could detect as few as 102 spores in both pure and crude DNA samples extracted from artificially inoculated tomato leaves, whereas PCR detected no fewer than 104 spores in pure DNA samples, failing to achieve detection in crude extracts. In in vivo bioassays, qcLAMP successfully detected B. cinerea immediately following spore deposition on leaf tissues (0 h post-inoculation) in both pure and crude DNA extracts. In contrast, conventional PCR was successful only in pure DNA samples from leaves collected 2 days post-inoculation (dpi) and in crude extracts at 3 dpi, when symptoms were already evident. Evaluations of in-field samples demonstrated that qcLAMP detected B. cinerea in 100% of crude extracts from symptomatic tissues and in 45% of asymptomatic (latently infected) samples. Conversely, conventional PCR failed to detect the pathogen in crude extracts, regardless of the disease status (symptomatic or asymptomatic). The developed method can serve as a rapid, reliable, and user-friendly tool for the early detection of B. cinerea in the field, enabling prompt assessment of inoculum pressure, thereby facilitating timely management interventions.

Plants doi: 10.3390/plants15213460

Authors: Raffaella Brigante Laura Marconi Luca Regni

Accurate, non-destructive quantification of shoot development in plant micropropagation is essential for optimising culture protocols and improving throughput. This study evaluated the performance of four RGB vegetation indices, Excess Green (ExG), Green Leaf Index (GLI), Modified Green Red Vegetation Index (MGRVI), and Visible Atmospherically Resistant Index (VARI), for the automated segmentation of Rubus spp. shoots in close-range orthophotos acquired with a smartphone camera. Twelve vessels, each containing fifteen blackberry shoots, grown at four sucrose concentrations (0, 7.5, 15 and 30 g L−1) were analysed at both pixel and ROI (Region Of Interest) levels. Classification thresholds were determined by maximising the Youden index on the ROC curve, both as a single global threshold and as concentration-specific thresholds. GLI and ExG substantially outperformed MGRVI and VARI in all discriminability metrics (AUC-ROC = 0.991 and 0.988, respectively), with stable classification performance across all sucrose concentrations. Projected shoot cluster area estimates derived from the two best-performing indices were benchmarked against a geometrically explicit reference obtained from a colour-filtered 3D point cloud reconstructed by Structure-from-Motion (SfM) photogrammetry from the same image set. Strong agreement was observed between RGB-based and point-cloud-derived area estimates (R2=0.990 for GLI), with Intersection over Union (IoU) ≥0.84 for all vessels. Concentration-specific thresholds provided a modest improvement in in-sample regression fit over the single global threshold without substantially affecting geometric agreement. A separate external dataset of six vessels, with independently annotated masks, was subsequently used to assess threshold transferability (mean IoU 0.908 for GLI and 0.861 for ExG). The results indicate that, under the tested conditions, GLI and ExG applied to smartphone-acquired orthophotos provide a relatively low-cost, non-destructive approach for projected shoot cluster area extraction in Rubus spp. in vitro cultures. The study is primarily methodological: the relationship between the image-derived measurements and independent biological descriptors of shoot development or propagation performance was not assessed.

Plants doi: 10.3390/plants15213461

Authors: Zhiying Li Yin Min Htwe Dapeng Zhang Peng Shi Qun Yu Xiangman He Yong Wang

Coconut zygotic embryo germination supports seedling establishment through coordinated embryo growth, tissue differentiation, and nutrient acquisition. A distinctive feature is the cotyledonary haustorium, which develops within the endosperm and transfers stored reserves to the growing seedling. However, transcriptional changes linking early germination to haustorium maturation and their relationships with phytohormone dynamics remain poorly understood. Here, we show that integrated time-resolved RNA sequencing, transcription factor (TF) and co-expression analyses, functional enrichment, and phytohormone profiling across 1–8 weeks after germination (WAG), including shoot/root (SR) and haustorium (Ha) tissues, reveal distinct developmental programs. We identified 792 core differentially expressed genes (DEGs), which formed four expression clusters. Cluster 4 was associated with early embryo development and included transcription factor (TF) modules involving VP1, ABI4, ARF1, BBM2, TCP4, ERF003, GRF2, and GRF6. In contrast, Cluster 1 increased during haustorium development and was enriched in hormone signaling, flavonoid biosynthesis, and cutin, suberine, and wax biosynthesis. MIF1, HOX16, and CPC showed high co-expression network connectivity. Hormone profiling showed relatively high auxin- and cytokinin-related metabolites early, progressive abscisic acid (ABA) increases, elevated ABA, indole-3-acetic acid (IAA), jasmonic acid (JA), jasmonoyl-isoleucine (JA-Ile), and salicylic acid (SA) in 6WAG-Ha, and increased methyl jasmonate (MeJA) and salicylic acid glucoside (SAG) in 8WAG-Ha. These findings reveal transcription factors and phytohormones during zygotic embryo germination, providing new understanding and reference to genetic transformation and somatic embryogenesis in coconut and other palm crops.

Plants doi: 10.3390/plants15213459

Authors: Libing Gao Jianghao Wang Xinzhi Zhang Xiayang Wu Jianning Chang Bing Wang Shaoqing Guo

Biochar performance in alkaline, low-fertility soil depends on feedstock, pyrolysis temperature, and application rate, yet the effects of mining-affected Artemisia biomass remain poorly characterized. Artemisia (SBC) and maize straw (BC) biochars produced at 400, 500, and 600 °C were applied at 1% or 5% (w/w) in a 40-day ryegrass pot experiment. We measured plant growth, chlorophyll, antioxidant enzyme activities, plant tissue Fe, Mn, Cu, and Zn, and soil pH, organic carbon, enzyme activities, and diethylenetriaminepentaacetic acid (DTPA)-extractable micronutrients. At 1%, SBC600 increased plant height and root length by 60.98% and 43.10%, respectively, versus the unamended control; SBC500 increased total chlorophyll by 145.71%. Under L-SBC500, DTPA-extractable Fe, Mn, Cu, and Zn were 100.26%, 80.37%, 153.01%, and 342.86% higher than in the control. The 5% rate raised soil pH to 9.53 and attenuated several plant and soil responses. Converting this locally available mining-area weed into biochar provides a waste-to-resource strategy that simultaneously manages metal-bearing biomass and reshapes the soil micronutrient profile: the Artemisia biochar was intrinsically richer in Fe, Mn, and Cu but lower in Zn than the conventional maize straw biochar, revealing a feedstock-specific nutrient signature. These short-term, concentration-based results demonstrate clear treatment-dependent benefits, although they do not yet quantify the source or transfer of elements or establish long-term environmental safety.

Plants doi: 10.3390/plants15213458

Authors: Sajida Hamdy Kashtoh Tensangmu Lama Tamang Kwang-Hyun Baek

Various abiotic challenges, including severe drought, extreme temperatures, salinity, and heavy metal toxicity, substantially limit plant growth and agricultural sustainability on a global scale. Sustainable approaches that strengthen crop resilience while minimizing environmental impacts are therefore urgently needed. Plant growth-promoting rhizobacteria (PGPR), beneficial fungi, and nanoparticles have individually demonstrated considerable potential in improving plant growth, nutrient acquisition, antioxidant defense, and stress tolerance. PGPR enhance plant performance through phytohormone production, nutrient solubilization, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, and activation of systemic defense responses. Beneficial fungi augment water uptake, nutrient absorption, osmotic adjustment, and stress-responsive signaling pathways, whereas nanoparticles facilitate efficient nutrient delivery, enhance photosynthetic performance, and support plant adaptation under adverse environmental conditions. Recent studies have further highlighted the synergistic benefits of PGPR–fungi, fungi–nanoparticle, and PGPR–nanoparticle interactions in mitigating abiotic stress. However, integrated tripartite interactions among PGPR, beneficial fungi, and nanoparticles remain insufficiently explored. This review summarizes current knowledge regarding their individual and combined roles, discusses the underlying physiological and molecular mechanisms, identifies existing research gaps, and proposes future perspectives for developing climate-resilient sustainable agricultural systems.

Plants doi: 10.3390/plants15213454

Authors: Titi Thongkamngam Sukritta Anutrakunchai Bancha Wiangsamut

Damping-off disease threatens durian seedling production and can facilitate pathogen dissemination during transplanting. This study evaluated the biocontrol potential of Trichoderma sp. THR3 against Sclerotium rolfsii associated with damping-off of durian seedlings. Three isolates, SCO1, SCO2, and SCO3, were obtained from symptomatic seedlings and identified using morphological characteristics and internal transcribed spacer sequence analysis. In pathogenicity tests conducted on two-month-old durian seedlings, SCO1 was the most aggressive isolate and produced 100% disease incidence. In dual culture, Trichoderma sp. THR3 inhibited the mycelial growth of SCO1 by 88.7% at 7 days after inoculation. In the in planta experiment using one-year-old grafted durian plants, treatment with Trichoderma sp. THR3 resulted in a 50% numerical reduction in final disease severity caused by S. rolfsii SCO1 relative to the pathogen-inoculated control at 9 days after inoculation (DAI). Disease incidence was numerically lower in THR3-treated plants (60%) than in the pathogen-inoculated control (100%); however, this difference was not statistically significant (p > 0.05). These findings indicate the potential of Trichoderma sp. THR3 as a candidate biological control agent against S. rolfsii SCO1 in grafted durian nursery plants, although further replicated experiments are needed to confirm its efficacy under nursery and field conditions.

Plants doi: 10.3390/plants15213457

Authors: Riwen Fei Jiayi Liu Minghang Guo Hongmei Zhu Siyang Duan Jiansong You Xiaoyu Li

Exogenous abscisic acid (ABA) enhances freezing tolerance, yet its dose-specificity and metabolic trade-offs in ornamental woody species remain poorly characterized. Here, we investigated the effects of exogenous ABA (0, 10, 30, 50 mg·L−1) on Rhododendron obtusum ‘Fen Zhuanglou’ seedlings exposed to −4 °C for 5 days. Based on phenotypic screening, 10 mg·L−1 ABA was identified as the hormetic optimum, whereas 50 mg·L−1 failed to alleviate injury. Under freezing stress, optimal ABA treatment (10 mg·L−1) reduced malondialdehyde (MDA) content by 23.5% compared to untreated controls, while partially rescuing superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities by 40.0%, 29.2%, and 35.2%, respectively. Notably, ABA triggered a metabolic trade-off: soluble protein declined by 22.98% while soluble sugar increased by 6.05%, suggesting a reallocation of metabolic resources. Furthermore, pigment profiling suggested impaired chlorophyll catabolism in unprotected leaves—chlorophyll a and xanthophylls accumulated to 7-fold and 43-fold the levels of ABA-treated leaves—whereas ABA-treated leaves retained a higher carotenoid fraction (7-fold). These findings establish that ABA-mediated freezing tolerance operates through a metabolic trade-off rather than universal preservation, providing an evidence-based protocol (10 mg·L−1) for ornamental rhododendron production in cold regions.

Plants doi: 10.3390/plants15213456

Authors: Hui Hu Nana Li Yanbing Yan Yongsheng Cui Xiaolong Hou Pengfei Wu

In the context of global warming and declining forest ecosystem services, this study explores underplanting effects on Pinus massoniana stand biomass allocation and soil C–N–P stoichiometry to support sustainable plantation management in southern China. Two underplanting treatments were established: P. massoniana underplanted with Schima superba (denoted by PS), and P. massoniana underplanted with S. superba + Michelia macclurei + Castanopsis hystrix (denoted by PSMC). A pure P. massoniana stand was set as the control (CK). The tree height, DBH and soil C–N–P stoichiometric characteristics were measured, and we compared biomass distribution among the tree layer, understory vegetation layer and litter layer. The results showed that compared with CK and PS, the PSMC significantly promoted the height and DBH growth of P. massoniana and substantially increased the biomass of all tree (leaves, branches, stems, and roots), sustaining higher stand productivity. In contrast, underplanted S. superba inhibited the growth of P. massoniana. Both underplanting treatments significantly reduced understory vegetation biomass, with herbs dominating the understory layer in both treatments. Litter biomass was the lowest under the PS treatment and was significantly lower than others. Compared with CK, both the PS and PSMC treatments significantly increased soil C, N, and P contents as well as soil C:P and N:P ratios in the 0–20 cm and 20–40 cm soil layers. The DBH of P. massoniana was positively correlated with soil C, N, and P concentrations in the 0–20 cm and 20–40 cm soil layers, and it exhibited an extremely significant positive correlation with soil N concentration and soil N:P ratio in the 40–60 cm soil layer. Tree-layer biomass was significantly increased with soil P concentration and the N:P ratio in the 0–20 cm soil layer, and it showed an extremely significant positive correlation with soil C and N concentrations in the 0–20 cm and 20–40 cm soil layers, as well as with soil N concentration and the N:P ratio in the 40–60 cm soil layer. In addition, litter-layer biomass was significantly and positively correlated with soil P concentration in the 20–40 cm soil layer. This association occurs because mixed underplanted broadleaves capture phosphorus released by litter decomposition via their well-developed roots and redistribute P to subsoil, and subsoil P availability controls litter production by regulating tree nutrient status. In conclusion, compared with CK, PSMC significantly improved P. massoniana individual growth and tree-layer biomass, while decreasing understory vegetation biomass and raising the 40–60 cm soil N:P ratio. Deep-soil N:P ratio was positively correlated with tree-layer biomass, implying that PSMC supports high stand productivity.

Plants doi: 10.3390/plants15213455

Authors: Yu-Xuan Tian Syuan-You Lin

Glycine betaine (GB) is used as an abiotic-stress biostimulant, but the performance of repeated foliar applications in perennial fruit crops under commercial conditions remains poorly defined. We evaluated four application strategies delivering cumulative GB inputs of 0, 42.5, 59.8, or 99.7 kg·ha−1 to greenhouse-grown ‘Tainung No. 1’ passion fruit (Passiflora edulis × P. edulis f. flavicarpa) during the cool season. Greenhouse temperature, vegetative growth, leaf greenness, visible reproductive development, marketable yield, and fruit quality were assessed. During the treatment period, 59 h below 10 °C were recorded, predominantly at night (83.1%). The 42.5 and 59.8 kg·ha−1 strategies did not consistently improve crop performance relative to the control. The 99.7 kg ha−1 strategy was associated with lower main-vine length during crop development, although final main-vine length at 144 DAT did not differ significantly from the control. The first visible flower bud appeared at a significantly higher node under 99.7 kg·ha−1 than in the control (27.1 vs. 20.8). Its appearance was significantly later under 99.7 than under 42.5 kg·ha−1 (29.9 vs. 19.6 DAT), but did not differ significantly from the control (21.0 DAT). The 99.7 kg ha−1 strategy also reduced marketable fruit number and yield by 14.6% and 13.8%, respectively. Fruit dimensions, individual fruit mass, and most quality traits were not consistently affected by GB, whereas harvest date more consistently influenced fruit quality. Under the recorded fluctuating thermal conditions, the intensive GB strategy was unfavorable, supporting crop-specific validation before repeated use.

Plants doi: 10.3390/plants15213453

Authors: Rongchun Ye Jianming Luo Ziqing Sun Xinjie Long Hongyan Fu Naying Gao Chenxing Hao Ling Sheng Yunlin Cao Youhan Kong Xianfeng Ma Yan Jin Jian Han

Citrus canker, caused by Xanthomonas citri subsp. citri (Xcc), is commonly managed with copper-based formulations, yet the performance of individual products may differ between laboratory and plant assays. We compared 16 commercial copper-containing formulations in plate-diffusion and root-drench assays, and inhibition zones varied among products. Thiosen Copper, however, produced no visible inhibition zone on agar but suppressed Xcc growth after root application. In sweet orange, this suppression appeared earlier than with Bordeaux Mixture at the same nominal total Cu concentrations. Thiosen Copper raised the leaf chlorophyll index without a significant increase in total leaf Cu, whereas Bordeaux Mixture led to greater leaf Cu accumulation. In separate experiments with Poncirus trifoliata, the two formulations produced different growth and tissue Cu responses, and root treatment altered PtCOPT transcript levels. Transient overexpression of CsCOPT4 in sweet orange leaves was accompanied by lower Xcc populations. These results reveal formulation-specific responses and nominate CsCOPT4 as a candidate for further study.

Plants doi: 10.3390/plants15213452

Authors: Jianhong Xie Jing Wang Juan Xu Lei Cheng Yifan Zeng Qingjun Shu Yaping Wang Baozhong Duan

Polygonatum species are medicinal and edible plants whose polysaccharides (PSPs) have attracted interest for their potential anti-aging effects. This review synthesizes evidence on the structural characteristics, pharmacological activities, molecular mechanisms, and translational prospects of PSPs. Relevant literature was retrieved from Web of Science and the China National Knowledge Infrastructure using terms related to Polygonatum polysaccharides, aging, structural characterization, and bioactivity. PSPs vary in molecular weight, monosaccharide composition, glycosidic linkages, and branching patterns. Studies using cells, Caenorhabditis elegans, and mammals report aging-related benefits associated with reduced oxidative stress and inflammation, modulation of gut microbiota and immune homeostasis, improved mitochondrial function, and metabolic regulation. Reported molecular changes were associated with Nrf2/HO-1, SIRT1/SIRT3, AMPK/PGC-1α, ERK/NF-κB, and DAF-16-related signaling. Although comparisons among PSP fractions suggest associations between structural features and biological activity, causal structure–activity relationships remain unconfirmed. Moreover, most evidence is preclinical, and differences in botanical sources, extraction methods, and polysaccharide characterization limit cross-study comparisons. Future research should prioritize standardized, structurally defined preparations, direct structure–activity testing, pharmacokinetics, long-term safety, and clinical validation to clarify the potential of PSPs for healthy aging.

Plants doi: 10.3390/plants15213451

Authors: Hua Yin Mingyu Zeng Ziwei Song Yinglong Wang Ziyi Liao Yebao Wu Li Li Juan Liu Ke Wang Anping Xiong

Accurate detection of emerged seedlings is essential for intelligent management in the context of industrialized pepper seeding cultivation. Manual assessment of seedling emergence rates is inefficient and subjective. Meanwhile, dense seedling distributions and complex substrate backgrounds in plug trays often cause false positives and missed detections in vision-based systems. To address these issues, Adaptive-Dynamic-Spatial YOLO (ADS-YOLO), a compact detection model, was built based upon YOLO11n. First, an Adaptive Scale Encoding (ASE) module was introduced into the neck to preserve fine-grained spatial information through parallel variable-scale processing and fusion. Second, a Dynamic Scale Sequence Fusion (DYSF) module combined content-aware DySample upsampling with a scale-indexed three-dimensional feature representation to improve multiscale alignment and fusion. Third, a Spatial-Channel Position Attention (SCPA) module was used to emphasize seedling-related responses and suppress interference from substrate particles, water stains, and reflections. On the pepper seedling validation set obtained from a fixed 8:1:1 image-level data split, ADS-YOLO achieved a precision of 79.0%, a recall of 75.5%, and an mAP@0.5 of 78.1%, representing improvements of 3.3, 3.2, and 2.9 percentage points over YOLO11n. The model contained approximately 3.04 M parameters and required 8.1 GFLOPs. Furthermore, a prototype based on offline video-frame analysis was developed to display seedling detections and frame-level counts. The prototype achieved an offline processing throughput of 24.5 FPS and a per-frame end-to-end processing time of 40.8 ms. These measurement results indicate that the method proposed in this paper can be used in practical scenarios, and also lay the foundation for future real-time tracking and counting.

Plants doi: 10.3390/plants15213449

Authors: Xiaodan Xu Zihan Zhang Ningning Sun Sian Peng Tangyi Chen Yanmin Qie Lige Geng Shihui Wang Weina Sun Yongxia Guo Jidao Du

Adzuki bean (Vigna angularis) is an important pulse crop, and rust disease severely restricts its production. Identifying resistance genes is essential for accelerating the development of rust-resistant cultivars through marker-assisted breeding. In this study, genotyping-by-sequencing (GBS) was performed on 178 adzuki bean accessions, generating 124.87 Gb of sequencing data and yielding 12,153 high-quality single-nucleotide polymorphism (SNP) markers with an average density of 26.56 SNPs/Mb across the genome. Principal component analysis (PCA) and K-means clustering divided the 178 accessions into three distinct genetic clusters, with the first five principal components explaining 31.90% of the total genetic variation. Population structure analysis based on these SNPs revealed that the cross-entropy criterion reached its minimum at K = 7, partitioning the panel into seven genetic subpopulations (C1–C7). A neighbor-joining phylogenetic tree further resolved the accessions into five major groups. Rust resistance was assessed at the seedling stage under greenhouse conditions with artificial inoculation of Uromyces vignae isolate ZXL01, and at the adult stage under natural field infection at a single experimental site. Genome-wide association analysis (GWAS) was conducted separately for the two assessments and identified fifteen SNPs significantly associated with rust resistance. Ten SNPs were identified at the seedling stage on chromosomes 1 and 3, while the remaining five SNPs were detected at the adult stage on chromosome 9. The SNPs associated with adult-stage rust resistance were converted into Kompetitive Allele-Specific PCR (KASP) markers, which were subsequently validated in an independent panel. The validation confirmed that the resistance-associated alleles were predominantly detected in highly resistant accessions, suggesting that these markers have potential utility for the preliminary screening of rust-resistant germplasm, although further breeding validation is required.

Plants doi: 10.3390/plants15213450

Authors: Hanlin Liu Xiaoshuo Jiang Chang Lu Yizeng Lu Dan Liu Xin Han Limin Sun

Quercus acutissima is an important tree species of coastal shelterbelts in eastern China; however, soil salinization severely constrains its growth and ecological benefits. Funneliformis mosseae can enhance plant stress resistance, yet the mechanisms by which it regulates salt tolerance in Quercus acutissima remain unclear. In this study, a two-factor pot experiment was conducted on half-sib families of Quercus acutissima with four salinity gradients (0‰, 2‰, 4‰, and 6‰, corresponding to 0%, 0.2%, 0.4%, and 0.6% (w/w) NaCl, respectively) and four inoculant doses (0, 10, 20, and 30 g), with three biological replicates per treatment, and the regulatory pathways were dissected by combining physiological measurements with transcriptome sequencing of functional leaf tissues collected from four selected treatment groups (the uninoculated control, mycorrhizal inoculation alone, salt stress alone, and salt stress combined with mycorrhizal inoculation). The results showed that inoculation with Funneliformis mosseae under salt-free conditions induced a slight stress-like response in Quercus acutissima; inoculation enhanced salt tolerance only under low-salt conditions (≤2‰ NaCl, mass concentration), with the 30 g high-dose inoculant being the most effective; salt stress promoted excessive root lignification and xylem embolism, whereas symbiosis with Funneliformis mosseae alleviated this damage. Successful root colonization by Funneliformis mosseae was established in all inoculated treatments, confirming the formation of a functional mycorrhizal symbiosis under both salt-free and salt-stressed conditions. The pentose phosphate pathway, the inositol phosphate–Ca2+ signaling pathway, and multiple hormone-related pathways coordinately regulated salt tolerance. This study provides a theoretical basis for cultivating Quercus acutissima in salinized soils, offers practical support for afforestation of Quercus acutissima on saline lands in the Yellow River Delta, and contributes to improving the quality and efficiency of coastal shelterbelts.

Plants doi: 10.3390/plants15213448

Authors: Muhammet Öner Cuma Akıncı

Water scarcity is a recurrent constraint to durum wheat production in semi-arid Mediterranean-type environments, yet genotype performance can change sharply with the timing and severity of seasonal drought. This study quantified phenological, chlorophyll-related, yield-component, and grain-yield responses of 45 durum wheat genotypes grown during 2021–2022 and 2022–2023 under rainfed and supplementary-irrigated field conditions in Diyarbakır, southeastern Türkiye. The experiment comprised equal-sized plots with four replications under each water regime. Supplementary irrigation was delivered by drip irrigation in five applications beginning at stem elongation. Precipitation from sowing to heading was 187.0 mm in 2021–2022 and 285.5 mm in 2022–2023, and April was warmer in the first season (17.9 °C versus 14.7 °C). Grain yield was significantly affected by growing season, water regime, genotype, and all two- and three-way interactions (p < 0.01), although diagnostic tests indicated heterogeneous residual variance among environments and the combined fixed-effect ANOVA is therefore interpreted cautiously. Mean grain yield under rainfed conditions was 466.5 and 3254.3 kg ha−1 in the first and second seasons, respectively, compared with 4208.1 and 5560.9 kg ha−1 under supplementary irrigation. The exceptionally low first-season rainfed yield represents a near-crop-failure environment; stand-count data were not available to separate establishment losses from subsequent drought effects. Pooled drought indices descriptively ranked Fırat-93 highest for STI (0.638) and GMP (3901.4 kg ha−1), whereas Şahinbey had the highest pooled YSI (0.518); these index differences were not subjected to inferential significance testing and should not be interpreted as statistically distinct tolerance classes. Grain yield was associated with SPAD and yield components in pooled genotype means, but these associations were environment dependent. The results characterize genotype responses across four site-year × water-regime combinations and provide hypotheses for validation in broader multi-location and multi-year trials.

Plants doi: 10.3390/plants15213447

Authors: Dan Jackson Carmen Mata Nadine Field Lorena Pizarro Adi Avni Stephen C. Whisson Keith Miller Stanley W. Botchway Charlotte Pain Verena Kriechbaumer

The endoplasmic reticulum (ER) is a dynamic membrane network that plays essential roles in protein secretion, lipid biosynthesis and cellular signalling. Its architecture is maintained by conserved ER-shaping proteins, including the recently identified Lunapark (LNP) family, which have primarily been characterised in Arabidopsis thaliana. Whether these mechanisms are conserved in crop species remains unknown. Here, we identify and characterise two tomato (Solanum lycopersicum) LNP proteins, SlyLNP1 and SlyLNP2, and investigate their roles in ER organisation and function. Both proteins localised to the ER but displayed distinct distribution patterns, with SlyLNP1 preferentially associated with cisternae and SlyLNP2 localising to both cisternae and tubules. Overexpression of either protein promoted cisternal expansion and altered ER network organisation. FRET-FLIM analyses demonstrated that both SlyLNP proteins interact with themselves, each other, the ER fusogen RHD3 and the membrane-curvature protein RTN1, indicating conservation of the ER-shaping machinery. Furthermore, SlyLNP overexpression reduced ER-to-Golgi trafficking, linking ER remodelling with secretory pathway function. In contrast to their Arabidopsis counterparts, however, expression of tomato LNP proteins in Nicotiana benthamiana did not reduce infection by Phytophthora infestans. Together, our findings demonstrate that the molecular mechanisms governing ER architecture are highly conserved in tomato while revealing that the physiological consequences of Lunapark-mediated ER remodelling have diverged between plant species.

Plants doi: 10.3390/plants15213446

Authors: Carlos Frey José Luis Acebes

Many of the processes studied in plant physiology, including stomatal regulation, responses to drought, formation of graft unions, interactions with pathogens, floral thermogenesis and freezing, are dynamic but difficult for students to observe directly. Consequently, teaching these topics often relies on indirect measurements or theoretical explanations, which makes it difficult for students to understand the link between physiological concepts and observable plant behaviour. Infrared thermography offers a rapid, non-invasive way to visualise temperature patterns that reflect underlying physiological processes. This article presents ten inquiry-based laboratory and field activities that integrate infrared thermography into undergraduate plant physiology teaching. Rather than focusing on the technology itself, the activities are organised around fundamental physiological concepts, challenging students to formulate hypotheses, interpret thermal images and relate temperature patterns to plant function. The activities cover topics such as water relations and transpiration, graft union formation, plant–pathogen interactions, drought detection, host–parasitic plant relationships, floral thermogenesis, floral cooling, ice nucleation, seed vigour assessment and responses to abiotic and biotic stresses. Beyond illustrating individual physiological phenomena, the activities emphasise the unifying principle that infrared thermography does not simply measure temperature but provides an integrated window into plant physiology by revealing processes that would otherwise be invisible. At the same time, students become familiar with a research tool that is increasingly used in plant science, precision agriculture and crop phenotyping. As infrared cameras become more affordable and accessible, thermography represents a powerful and realistic approach to enriching inquiry-based learning and strengthening the connection between undergraduate teaching and contemporary plant research.

Plants doi: 10.3390/plants15213445

Authors: Adeel Abbas Rashida Hameed Sidra Balooch Shimeng Yang Aitezaz Ali Shahani Usman Zulfiqar Mohammed Zari

This study investigated the effects of rice straw biochar produced at 450 °C (RBC450) and 700 °C (RBC700) on Pb immobilization, soil properties, and the physiological and molecular responses of Wedelia trilobata (L.) grown in Pb-contaminated soil. Biochar was applied at 1%, 2.5%, and 5% (w/w) to soil contaminated with 150 mg kg−1 Pb. Changes in soil physicochemical properties, Pb availability and accumulation in plant tissues, physiological performance, oxidative stress indicators, antioxidant enzyme activities, and antioxidant-related gene expression were evaluated. Both biochars reduced soil-available Pb and Pb accumulation in roots and shoots, with RBC700 showing greater Pb immobilization. Biochar application also improved soil pH, electrical conductivity, organic carbon, nitrate nitrogen, and available phosphorus. In plants, biochar enhanced catalase and peroxidase activities while reducing malondialdehyde accumulation and reactive oxygen species production, indicating alleviation of Pb-induced oxidative stress. qRT-PCR analysis further showed modulation of SOD-, POD-, and CAT-related antioxidant responses under Pb stress, with RBC450 treatment strengthening antioxidant defense at the transcriptional level. Overall, rice straw biochar reduced soil-extractable Pb and modified soil physicochemical properties and antioxidant responses in W. trilobata, with RBC700 showing greater effectiveness in decreasing Pb extractability. However, these changes did not consistently restore all physiological parameters under Pb stress.

Plants doi: 10.3390/plants15213444

Authors: Hanjian Hu Lingchao He Rujie Ye Kang Xu

Multiple morphological factors of desert plants were employed to construct aboveground biomass (AGB) estimation models in the Gobi Desert, located in the northwestern part of Gansu Province, China, where gravel surfaces account for more than half the area. At the individual scale, estimation models incorporating variables related to canopy cover accurately estimated AGB (R2 = 0.623~0.848). At the quadrat scale, the cover-allometric model effectively estimated multi-species AGB (AGB = 0.302 × Ct0.900, R2 = 0.725). At the regional scale, the model incorporating relative canopy cover was effective for estimating aboveground biomass density (AGBD) and could be combined with regional area estimates to determine the AGB of desert plants at any spatial scale (AGBD = 20.114 × Cr1.043, R2 = 0.8313). AGB estimation models based on canopy cover not only overcome scale limitations but also exhibit a tendency toward improved model fit with increasing scales. However, when applied to remote-sensing data, the model exhibits relatively low fitting accuracy (AGBD = 4.398 × Cr0.538, R2 = 0.3656). It demonstrates good stability when applied to Gobi Desert habitats similar to the study area, but its applicability in other arid regions still requires further validation.

Plants doi: 10.3390/plants15213443

Authors: Klaudija Carović-Stanko Boris Lazarević Tomislav Javornik Monika Vidak Kristina Kljak

Intercropping can affect nutrient availability and plant biochemical composition, but information on these effects in medicinal and aromatic plants is still limited. This study evaluated the effects of intercropping with common bean (Phaseolus vulgaris L.) on mineral composition, carotenoids, and tocopherols in two basil (Ocimum basilicum L.) cultivars, ‘Genovese’ and ‘Sweet basil’, grown at different basil–bean ratios. Intercropping significantly affected all analyzed minerals, although the response differed between cultivars for several elements. ‘Genovese’ showed increases in Zn, Mn, and Cu, while Zn content in ‘Sweet basil’ remained relatively stable. In contrast, N, P, and Fe concentrations decreased in some treatments. Total carotenoids and total lutein decreased under intercropping in both cultivars, whereas β-carotene responses were less consistent and depended on cultivar and intercropping ratio. In contrast, total tocopherols and α-tocopherol generally increased under intercropping. Zn, Mn, and Cu were negatively correlated with several carotenoids but positively correlated with tocopherols. PCA separated monocropped from most intercropped samples and showed that both mineral and antioxidant variables contributed to sample differentiation. These findings show that the compositional response of basil to common bean intercropping depends on cultivar, intercropping ratio, and the group of compounds considered.

Plants doi: 10.3390/plants15213442

Authors: Changli Mao Xia Zeng Tian Yang Chenwanli Li Cheng Zheng Junming Ai Xiaodong Liu Jing Zhang Xiaoping Zhu Haoqi Shi Yu Wu Jin Liu

Natural rubber from Hevea brasiliensis serves as an irreplaceable industrial raw material. Whether rubber-yield potential is constrained by sucrose-sink sucrose partitioning remains incompletely understood. Here, we integrated transcriptomics and targeted sugar metabolomics across leaves, bark and latex from three H. brasiliensis clones with contrasting yields (designated H, M, and L). Transcriptomic profiling revealed tissue-specific functional differentiation: leaves were enriched in photosynthetic carbon fixation, bark in substance transport, and latex in defence and regeneration. The H clone did not show enhanced transcriptional enrichment of photosynthetic pathways relative to the L clone. Instead, it displayed a higher transcript abundance of invertase and sucrose synthase genes alongside elevated hexose levels, pointing to putative genotypic differences in sucrose hydrolysis and hexose turnover. Metabolomic profiling revealed greater sucrose accumulation in leaves of the L clone, accompanied by relatively low sucrose concentrations in latex. WGCNA analysis identified the MEpink module as strongly correlated with leaf sucrose content. This module is enriched in thiamine and cyanoamino acid metabolism and contains candidate transcription factors WRKY40, MYC2 and NAC002. Our results suggest that yield variation among these clones correlates with distinct sucrose distribution and utilization profiles. These multi-omics signatures provide a molecular resource to support high-yield rubber tree breeding.

Plants doi: 10.3390/plants15213441

Authors: Emre Yazar Bülent Akgün Emre Babur Sinan Kordon Erdi Ekren

Türkiye holds the world’s largest natural stands of Lebanon cedar (Cedrus libani A. Rich.) and extensive Crimean juniper forest (Juniperus excelsa M. Bieb.), together comprising 884,361 ha of closed, drought-exposed forest. Both assimilate at needle water potentials near −3.0 MPa, yet lose part of their potential assimilation to photorespiration and to stomatal closure under water deficit. A literature-parameterized scenario model relaxes both constraints against a radiation-limited ceiling. The zonal photorespiratory and stomatal input fractions are structured expert estimates, not measurements, which are: 8–16% and 14–28% of potential gross primary production (GPP), or 13–23% and 11–23% after coupling. Together they reduce weighted-mean net primary productivity from 3.29 to 2.17 Mg C ha−1 yr−1, a 34.1% reduction in potential GPP (Monte Carlo interval 31.0–36.4%, conditional on the elicitation). Of that loss, 39–46% is obligate photorespiration beyond management reach, depending on how the jointly caused interaction is assigned; the obligate share falls along the aridity gradient. Radial growth in these species occupies a narrow climatic window that does not coincide with peak assimilation, so forgone wood is reported as a function of the source-control coefficient φ, with φ = 1 as an upper bound. At φ = 1, the estate forgoes up to 1.5 million m3 yr−1 of stemwood, and the 3.6 million Mg yr−1 of unfixed carbon (as CO2-equivalent, not net sequestration) is worth, together with that wood, about US$ 0.20 billion; at φ = 0.5 the wood and the value halve, and at φ = 0 no wood is forgone.