Plants, Volume 12, Issue 11 (June-1 2023) – 165 articles

Sardinia and Corsica are two Mediterranean islands where the genus Armeria is represented by 11 taxa, 10 out of which are endemic. An integrative approach, using molecular phylogeny, karyology, and seed and plant morphometry was used to resolve the complex taxonomy and systematics in this group. We found that several taxa are no longer supported by newly produced data. Accordingly, we describe a new taxonomic hypothesis that only considers five species: Armeria leucocephala and A. soleirolii, endemic to Corsica, and A. morisii, A. sardoa, and A. sulcitana, endemic to Sardinia. Full article

Despite advances in vaccine development, influenza remains a persistent global health threat and the search for a broad-spectrum recombinant vaccine against influenza continues. The extracellular domain of the transmembrane protein M2 (M2e) of the influenza A virus is highly conserved and can be used to develop a universal vaccine. M2e is a poor immunogen by itself, but it becomes highly immunogenic when linked to an appropriate carrier. Here, we report the transient expression of a recombinant protein comprising four tandem copies of M2e fused to an artificial self-assembling peptide (SAP) in plants. The hybrid protein was efficiently expressed in Nicotiana benthamiana using the self-replicating potato virus X-based vector pEff. The protein was purified using metal affinity chromatography under denaturing conditions. The hybrid protein was capable of self-assembly in vitro into spherical particles 15–30 nm in size. The subcutaneous immunization of mice with M2e-carrying nanoparticles induced high levels of M2e-specific IgG antibodies in serum and mucosal secretions. Immunization provided mice with protection against a lethal influenza A virus challenge. SAP-based nanoparticles displaying M2e peptides can be further used to develop a recombinant “universal” vaccine against influenza A produced in plants. Full article

Alfalfa (Medicago satiua L.) is a major forage legume in semi-arid regions such as North China Plain and is the material foundation for the development of herbivorous animal husbandry. How to improve the yield of alfalfa per unit area from a technical perspective and achieve high-yield cultivation of alfalfa is the focus of research by scientific researchers and producers. To evaluate the effects of irrigation and P fertilization as well as the P residual effect on alfalfa yield, we conducted a six-year (2008–2013) field experiment in loamy sand soil. There were four irrigation levels (W0: 0 mm, W1: 25 mm, W2: 50 mm, W3: 75 mm per time, four times a year) and three P fertilization levels (F0: 0 kg P₂O₅ ha⁻¹, F1: 52.5 kg P₂O₅ ha⁻¹, F2: 105 kg P₂O₅ ha⁻¹ per time, twice a year). The highest dry matter yield (DMY) was obtained in the W2F2 treatment, with an annual mean of 13,961.1 kg ha⁻¹. During 2009–2013, the DMY of first and second-cut alfalfa increased significantly with increasing irrigation levels, whereas the opposite pattern was observed in fourth-cut alfalfa. Regression analysis revealed that the optimal amount of water supply (sum of seasonal irrigation and rainfall during the growing season) to obtain maximum DMY was between 725 and 755 mm. Increasing P fertilization contributed to significantly higher DMY in each cut of alfalfa during 2010–2013 but not in the first two growing seasons. The mean annual DMY of W0F2, W1F2, W2F2, and W3F2 treatments was 19.7%, 25.6%, 30.7%, and 24.1% higher than that of W0F0 treatment, respectively. When no P fertilizer was applied in F2 plots in 2013, soil availability and total P concentrations, annual alfalfa DMY, and plant nutrient contents did not differ significantly compared with those in fertilized F2 plots. Results of this study suggest that moderate irrigation with lower annual P fertilization is a more environmentally sound management practice while maintaining alfalfa productivity in the semi-arid study area. Full article

Throughout evolution, plants have developed a highly complex defense system against different threats, including phytopathogens. Plant defense depends on constitutive and induced factors combined as defense mechanisms. These mechanisms involve a complex signaling network linking structural and biochemical defense. Antimicrobial and pathogenesis-related (PR) proteins are examples of this mechanism, which can accumulate extra- and intracellular space after infection. However, despite their name, some PR proteins are present at low levels even in healthy plant tissues. When they face a pathogen, these PRs can increase in abundance, acting as the first line of plant defense. Thus, PRs play a key role in early defense events, which can reduce the damage and mortality caused by pathogens. In this context, the present review will discuss defense response proteins, which have been identified as PRs, with enzymatic action, including constitutive enzymes, β-1,3 glucanase, chitinase, peroxidase and ribonucleases. From the technological perspective, we discuss the advances of the last decade applied to the study of these enzymes, which are important in the early events of higher plant defense against phytopathogens. Full article

Rice is a crucial food crop, but it is frequently affected by diseases during its growth process. Some of the most common diseases include rice blast, flax leaf spot, and bacterial blight. These diseases are widespread, highly infectious, and cause significant damage, posing a major challenge to agricultural development. The main problems in rice disease classification are as follows: (1) The images of rice diseases that were collected contain noise and blurred edges, which can hinder the network’s ability to accurately extract features of the diseases. (2) The classification of disease images is a challenging task due to the high intra-class diversity and inter-class similarity of rice leaf diseases. This paper proposes the Candy algorithm, an image enhancement technique that utilizes improved Canny operator filtering (the gravitational edge detection algorithm) to emphasize the edge features of rice images and minimize the noise present in the images. Additionally, a new neural network (ICAI-V4) is designed based on the Inception-V4 backbone structure, with a coordinate attention mechanism added to enhance feature capture and overall model performance. The INCV backbone structure incorporates Inception-iv and Reduction-iv structures, with the addition of involution to enhance the network’s feature extraction capabilities from a channel perspective. This enables the network to better classify similar images of rice diseases. To address the issue of neuron death caused by the ReLU activation function and improve model robustness, Leaky ReLU is utilized. Our experiments, conducted using the 10-fold cross-validation method and 10,241 images, show that ICAI-V4 has an average classification accuracy of 95.57%. These results indicate the method’s strong performance and feasibility for rice disease classification in real-life scenarios. Full article

Using in situ near-surface observations of solar-induced chlorophyll fluorescence (SIF) and gross primary productivity (GPP) of a subtropical evergreen coniferous forest in southern China, this study analyzed the dynamics of SIF, GPP and their environmental responses, and explored the potential of SIF in characterizing the variation of GPP. The results showed that SIF and GPP have similar diurnal and seasonal variation and both reach the highest value in summer, indicating that the SIF can be applied to indicate the seasonal variation of GPP for the subtropical evergreen co-niferous. With the increase in temporal scale, the correlation between SIF and GPP becomes more linear. The diurnal variations of both SIF and GPP were characterized by photosynthetically active radiation (PAR), the seasonal variations of SIF and GPP were influenced by air temperature (Ta) and PAR. Probably due to the absent of drought stress during the study period, no significant correlation was detected between soil water content (SWC) and either SIF or GPP. With the in-crease in Ta, PAR or SWC, the linear correlation between the SIF and GPP gradually decreased, and when Ta or PAR was relatively higher, the correlation between SIF and GPP become weakly. Further research is still needed to illustrate the relationship between SIF and GPP under drought condition which occurred frequently in this region based on longer observation. Full article

A study of the distribution of orchid species in Puglia, based on an analysis of 2084 bibliographic reports from 2000 to 2022, was carried out with the aim of revising and updating the information on the consistency of the Orchidaceae family in Puglia, with a special focus on assessing threatened species occurring inside and outside protected areas. The work presents a checklist of the Orchidaceae taxa (genera, species, and subspecies) found in the region, including observations on genera and species that present taxonomic challenges. A total of 113 taxa (i.e., species and subspecies), distributed across 16 genera, are listed in alphabetical order. The most representative genera were Ophrys (51 taxa), Serapias (15 taxa), and Epipactis (11 taxa). Additionally, 49 taxa (43.4%) were found to be endemic to Italy, with 21 of these, mostly belonging to the Ophrys genus, being exclusive to Puglia. Our study notes two different trends of distribution: a predominantly coastal distribution for orchid records located in southern Puglia (the Salento peninsula) and a more widespread distribution for the other provinces. Our study also shows that the greatest number of records locate orchids in protected areas with a positive correlation between their presence and habitats cited in Directive 92/43/EEC. Full article

Notorious invasive Bohemian knotweed Reynoutria × bohemica Chrtek et Chrtková is a hybrid of two species, Reynoutria japonica Houtt. and Reynoutria sachalinensis (F. S. Petrop.) Nakai in T. Mori which spontaneously developed in Europe, outside the natural distribution of its parental species. Its success could potentially lie in its allelopathic activity, which was confirmed in a number of experiments conducted with the leaf and root exudates, testing their effect on the germination and growth of various test plants. Here, we tested its allelopathic potential using different concentrations of leaf exudates on two test plants, Triticum aestivum L. and Sinapis alba L., in Petri dishes and pots with soil and by growing test plants in the soil sampled in knotweed stands on the edges of stands and outside of stands. Tests in Petri dishes and pots with soil to which leaf exudates were added have shown a decrease in germination and growth in comparison to the control, hence confirming the allelopathic effect. However, this was not confirmed in a test with in situ soil samples, where no statistically significant differences were observed, neither in the growth of test plants nor in the chemical characteristics (pH, soil organic matter, humus content) of the soil. Therefore, the persistence of Bohemian knotweed at already invaded sites could be attributed to its efficient use of resources (light and nutrients) through which it outcompetes native plants. Full article

Water deficit is a significant environmental stress that has a negative impact on plant growth and yield. In this research, the positive significance of kaolin and SiO₂ nanoparticles in moderating the detrimental effects of water deficit on maize plant growth and yield is investigated. The foliar application of kaolin (3 and 6%) and SiO₂ NPs (1.5 and 3 mM) solutions increased the growth and yield variables of maize plants grown under normal conditions (100% available water) and drought stress conditions (80 and 60% available water (AW)). In addition, plants treated with SiO₂ NPs (3 mM) demonstrated increased levels of important osmolytes, such as proline and phenol, and maintained more of their photosynthetic pigments (net photosynthetic rate (PN), stomatal conductance (gs), intercellular CO₂ concentration (Ci), and transpiration rate (E)) than with other applied treatments under either stress or non-stress conditions. Furthermore, the exogenous foliar application of kaolin and SiO₂ NPs also reduced the amounts of hydroxyl radicals (OH), superoxide anions (O₂), hydrogen peroxide (H₂O₂), and lipid peroxidation in maize plants experiencing a water deficit. In contrast, the treatments led to an increase in the activity of antioxidant enzymes such as peroxidase (POX), ascorbate peroxidase (APX), glutathione peroxidase (GR), catalase (CAT), and superoxide dismutase (SOD). Overall, our findings indicate the beneficial impact of the application of kaolin and silicon NPs, particularly the impact of SiO₂ NPs (3 mM) on managing the negative, harmful impacts of soil water deficit stress in maize plants. Full article

The plant hormone ABA (abscisic acid) is able to regulate plant responses to abiotic stresses via regulating the expression of ABA response genes. BIC1 (Blue-light Inhibitor of Cryptochromes 1) and BIC2 have been identified as the inhibitors of plant cryptochrome functions, and are involved in the regulation of plant development and metabolism in Arabidopsis . In this study, we report the identification of BIC2 as a regulator of ABA responses in Arabidopsis . RT-PCR (Reverse Transcription-Polymerase Chain Reaction) results show that the expression level of BIC1 remained largely unchanged, but that of BIC2 increased significantly in response to ABA treatment. Transfection assays in Arabidopsis protoplasts show that both BIC1 and BIC2 were mainly localized in the nucleus, and were able to activate the expression of the co-transfected reporter gene. Results in seed germination and seedling greening assays show that ABA sensitivity was increased in the transgenic plants overexpressing BIC2, but increased slightly, if any, in the transgenic plants overexpressing BIC1. ABA sensitivity was also increased in the bic2 single mutants in seedling greening assays, but no further increase was observed in the bic1 bic2 double mutants. On the other hand, in root elongation assays, ABA sensitivity was decreased in the transgenic plants overexpressing BIC2, as well as the bic2 single mutants, but no further decrease was observed in the bic1 bic2 double mutants. By using qRT-PCR (quantitative RT-PCR), we further examined how BIC2 may regulate ABA responses in Arabidopsis , and found that inhibition of ABA on the expression of the ABA receptor genes PYL4 (PYR1-Like 4) and PYL5 were decreased, but promotion of ABA on the expression of the protein kinase gene SnRK2.6 (SNF1-Related Protein Kinases 2.6) was enhanced in both the bic1 bic2 double mutants and 35S:BIC2 overexpression transgenic plants. Taken together, our results suggest that BIC2 regulates ABA responses in Arabidopsis possibly by affecting the expression of ABA signaling key regulator genes. Full article

In hazelnut, foliar nutrition is utilized globally to integrate microelement deficiencies and optimize their assimilation and effects on yield performances. Nevertheless, nut quality and kernel composition can be positively affected by foliar nutrition. Recently, several studies pointed out the need for increasing the sustainability of orchard nutrition by proposing the management of not only micronutrients, but also main components, such as nitrogen, through foliar spraying. In our study, different foliar fertilizers were used to understand the effectiveness of supporting hazelnut productivity and nut and kernel quality. Water was used as a control. Foliar fertilizations affected tree annual vegetative growth, improved kernel weight and decreased the incidence of blanks compared to the control. Differences in fat, protein, and carbohydrate concentration were also found among treatments, with increased fat concentrations and total polyphenols content in fertilized treatments. Foliar fertilization improved the oil composition of the kernels, though fatty acid composition responded differently to nutrients spray. Oleic acid concentration was promoted, while palmitic acid concentration was reduced in fertilized plants compared to control trees. Furthermore, CD and B trees were characterized by an increase in the ratio of unsaturated/saturated fatty acids compared to untreated trees. Finally, foliar spraying improved lipid stability compared to the control due to higher total polyphenol concentration. Full article

MADS-box is a vital transcription factor family that functions in plant growth and development. Apart from APETALA2, all genes in the ABCDE model that explain the molecular mechanism of floral organ development belong to the MADS-box family. Carpel and ovule numbers in plants are essential agronomic traits that determine seed yield, and multilocular siliques have great potential for the development of high-yield varieties of Brassica. In this study, ABCDE genes in the MADS-box family from Brassica rapa were identified and characterized. Their tissue-specific expression patterns in floral organs and their differential expression in different pistil types of B. rapa were revealed by qRT-PCR. A total of 26 ABCDE genes were found to belong to the MADS-box family. Our proposed ABCDE model of B. rapa is consistent with that of Arabidopsis thaliana, indicating that ABCDE genes are functionally conserved. These results of qRT-PCR showed that the expression levels of class C and D genes were significantly different between the wild-type (wt) and tetracarpel (tetrac) mutant of B. rapa. Interestingly, the expression of the homologs of class E genes was imbalanced. Therefore, it is speculated that class C, D, and E genes are involved in developing the carpel and ovule of B. rapa. Our findings reveal the potential for the selection of candidate genes to improve yield traits in Brassica crops. Full article

Cassava witches’ broom disease (CWBD) is one of the main diseases of cassava in Southeast Asia (SEA). Affected cassava plants show reduced internodal length and proliferation of leaves (phyllody) in the middle and top part of the plant, which results in reduced root yields of 50% or more. It is thought to be caused by phytoplasma; however, despite its widespread distribution in SEA still little is known about CWBD pathology. The overarching goal of this study was to review and corroborate published information on CWBD biology and epidemiology considering recent field observations. We report the following: (1) CWBD symptoms are conserved and persistent in SEA and are distinct from what has been reported as witches’ broom in Argentina and Brazil. (2) In comparison with cassava mosaic disease, another major disease of cassava in SEA, symptoms of CWBD develop later. (3) Phytoplasma detected in CWBD-affected plants belong to different ribosomal groups and there is no association study available indicating phytoplasma as the causing agent of CWBD. These findings are essential clues for designing surveillance and management strategies and for future studies to better understand the biology, tissue localization and spatial spread of CWBD in SEA and other potential risk areas. Full article

Cannabis sativa L. is typically propagated through micropropagation or vegetative cuttings, but the use of root-inducing hormones, such as indole-3-butyric acid (IBA), is not allowed for growing medicinal cannabis in Denmark. This study examined alternative rooting treatments, including Rhizobium rhizogenes inoculation, water-only as well as IBA treatments, in eight cannabis cultivars. PCR on root tissue suggested that 19% of R. rhizogenes-inoculated cuttings were transformed. These were derived from “Herijuana”, “Wild Thailand”, “Motherlode Kush”, and “Bruce Banner”, indicating a variation in cultivar susceptibility toward R. rhizogenes. A 100% rooting success was achieved regardless of cultivar and treatment, suggesting that alternative rooting agents are not required for efficient vegetative propagation. However, rooted cuttings differed in shoot morphology with improved shoot growth in cuttings treated with R. rhizogenes (195 ± 7 mm) or water (185 ± 7 mm) while inhibited shoot growth under IBA treatment (123 ± 6 mm). This could have advantageous economic implications should cuttings not treated with hormone reach maturity faster than those exposed to the hormone, thereby contributing to completing a full growing cycle more effectively. IBA exposure increased root length, root dry weight, and root/shoot dry weight ratio compared to cuttings treated with R. rhizogenes or water but simultaneously inhibited shoot growth compared to these. Full article

Duckweeds (Lemnaceae) are small, simply constructed aquatic higher plants that grow on or just below the surface of quiet waters. They consist primarily of leaf-like assimilatory organs, or fronds, that reproduce mainly by vegetative replication. Despite their diminutive size and inornate habit, duckweeds have been able to colonize and maintain themselves in almost all of the world’s climate zones. They are thereby subject to multiple adverse influences during the growing season, such as high temperatures, extremes of light intensity and pH, nutrient shortage, damage by microorganisms and herbivores, the presence of harmful substances in the water, and competition from other aquatic plants, and they must also be able to withstand winter cold and drought that can be lethal to the fronds. This review discusses the means by which duckweeds come to grips with these adverse influences to ensure their survival. Important duckweed attributes in this regard are a pronounced potential for rapid growth and frond replication, a juvenile developmental status facilitating adventitious organ formation, and clonal diversity. Duckweeds have specific features at their disposal for coping with particular environmental difficulties and can also cooperate with other organisms of their surroundings to improve their survival chances. Full article

Radish (Raphanus sativus) plants exhibit varied root colors due to the accumulation of chlorophylls and anthocyanins compounds that are beneficial for both human health and visual quality. The mechanisms of chlorophyll biosynthesis have been extensively studied in foliar tissues but remain largely unknown in other tissues. In this study, we examined the role of NADPH:protochlorophyllide oxidoreductases (PORs), which are key enzymes in chlorophyll biosynthesis, in radish roots. The transcript level of RsPORB was abundantly expressed in green roots and positively correlated with chlorophyll content in radish roots. Sequences of the RsPORB coding region were identical between white (948) and green (847) radish breeding lines. Additionally, virus-induced gene silencing assay with RsPORB exhibited reduced chlorophyll contents, verifying that RsPORB is a functional enzyme for chlorophyll biosynthesis. Sequence comparison of RsPORB promoters from white and green radishes showed several insertions and deletions (InDels) and single-nucleotide polymorphisms. Promoter activation assays using radish root protoplasts verified that InDels of the RsPORB promoter contribute to its expression level. These results suggested that RsPORB is one of the key genes underlying chlorophyll biosynthesis and green coloration in non-foliar tissues, such as roots. Full article

The Afromontane and Afroalpine areas constitute some of the main biodiversity hotspots of Africa. They are particularly rich in plant endemics, but the biogeographic origins and evolutionary processes leading to this outstanding diversity are poorly understood. We performed phylogenomic and biogeographic analyses of one of the most species-rich plant genera in these mountains, Helichrysum (Compositae-Gnaphalieae). Most previous studies have focused on Afroalpine elements of Eurasian origin, and the southern African origin of Helichrysum provides an interesting counterexample. We obtained a comprehensive nuclear dataset from 304 species (≈50% of the genus) using target-enrichment with the Compositae1061 probe set. Summary-coalescent and concatenation approaches combined with paralog recovery yielded congruent, well-resolved phylogenies. Ancestral range estimations revealed that Helichrysum originated in arid southern Africa, whereas the southern African grasslands were the source of most lineages that dispersed within and outside Africa. Colonization of the tropical Afromontane and Afroalpine areas occurred repeatedly throughout the Miocene–Pliocene. This timing coincides with mountain uplift and the onset of glacial cycles, which together may have facilitated both speciation and intermountain gene flow, contributing to the evolution of the Afroalpine flora. Full article

The common bean has received attention as a model plant for legume studies, but little information is available about the morphology of its pods and the relation of this morphology to the loss of seed dispersal and/or the pod string, which are key agronomic traits of legume domestication. Dehiscence is related to the pod morphology and anatomy of pod tissues because of the weakening of the dorsal and ventral dehiscence zones and the tensions of the pod walls. These tensions are produced by the differential mechanical properties of lignified and non-lignified tissues and changes in turgor associated with fruit maturation. In this research, we histologically studied the dehiscence zone of the ventral and dorsal sutures of the pod in two contrasting genotypes for the dehiscence and string, by comparing different histochemical methods with autofluorescence. We found that the secondary cell wall modifications of the ventral suture of the pod were clearly different between the dehiscence-susceptible and stringy PHA1037 and the dehiscence-resistant and stringless PHA0595 genotypes. The susceptible genotype had cells of bundle caps arranged in a more easily breakable bowtie knot shape. The resistant genotype had a larger vascular bundle area and larger fibre cap cells (FCCs), and due to their thickness, the external valve margin cells were significantly stronger than those from PHA1037. Our findings suggest that the FCC area, and the cell arrangement in the bundle cap, might be partial structures involved in the pod dehiscence of the common bean. The autofluorescence pattern at the ventral suture allowed us to quickly identify the dehiscent phenotype and gain a better understanding of cell wall tissue modifications that took place along the bean’s evolution, which had an impact on crop improvement. We report a simple autofluorescence protocol to reliably identify secondary cell wall organization and its relationship to the dehiscence and string in the common bean. Full article

This research aimed to optimize pressure (10–20 MPa) and temperature (45–60 °C) conditions for supercritical fluid extraction (SFE) of Makwaen pepper (Zanthoxylum myriacanthum) extract (ME) in comparison to conventional hydro-distillation extraction. Various quality parameters, including yield, total phenolic compounds, antioxidants, and antimicrobial activities of the extracts, were assessed and optimized using a central composite design. The optimal SFE conditions were found to be 20 MPa at 60 °C, which resulted in the highest yield (19%) and a total phenolic compound content of 31.54 mg GAE/mL extract. IC₅₀ values for DPPH and ABTS assays were determined to be 26.06 and 19.90 μg/mL extract, respectively. Overall, the ME obtained through SFE exhibited significantly better physicochemical and antioxidant properties compared to ME obtained through hydro-distillation extraction. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that beta-pinene was the major component in the ME obtained through SFE (23.10%), followed by d-limonene, alpha-pinene, and terpinen-4-ol at concentrations of 16.08, 7.47, and 6.34%, respectively. On the other hand, the hydro-distillation-extracted ME showed stronger antimicrobial properties than the SFE-extracted ME. These findings suggest that both SFE and hydro-distillation have the potential for extracting Makwaen pepper, depending on the intended purpose of use. Full article

Perilla leaves are known to be a rich source of polyphenols, which have been shown to exhibit various biological effects. This study aimed to compare the bioefficacies and bioactivities of fresh (PLE_f) and dry (PLE_d) Thai perilla (Nga-mon) leaf extracts. Phytochemical analysis indicated that both PLE_f and PLE_d were abundant in rosmarinic acid and bioactive phenolic compounds. PLE_d, which had higher levels of rosmarinic acid but lower levels of ferulic acid and luteolin than PLE_f, exhibited greater effectiveness in a free radical scavenging assay. Furthermore, both extracts were found to suppress intracellular ROS generation and exhibit antimutagenic activity against food-borne carcinogens in S. typhimurium. They also attenuated lipopolysaccharide-induced inflammation in RAW 264.7 cells by inhibiting the expression of nitric oxide, iNOS, COX-2, TNF-α, IL-1β, and IL-6 through the suppression of NF-κB activation and translocation. However, PLE_f exhibited a higher ability to suppress cellular ROS production and higher antimutagenic and anti-inflammatory activities than PLE_d, which can be attributed to its combination of phytochemical components. Overall, PLE_f and PLE_d have the potential to serve as natural bioactive antioxidant, antimutagenic, and anti-inflammatory agents to achieve potential health benefits. Full article

Gardenia jasminoides fruits are extensively grown worldwide, with a large harvest, and its major medicinal ingredients are geniposide and crocins. Research on their accumulation and biosynthsis-related enzymes is rare. In this study, the accumulation of geniposide and crocin of G. jasminoides fruits at different developmental stages were clarified by HPLC. The highest cumulative amount of geniposide was 2.035% during the unripe-fruit period, and the highest content of crocin was 1.098% during the mature-fruit period. Furthermore, transcriptome sequencing was performed. A total of 50 unigenes encoding 4 key enzymes related in geniposide biosynthsis pathways were screened, and 41 unigenes encoding 7 key enzymes in the pathways of crocin were elucidated. It was found that the expression levels of differentially expressed genes of DN67890_c0_g1_i2-encoding GGPS, which is highly related to geniposide biosynthesis, and DN81253_c0_g1_i1-encoding lcyB, DN79477_c0_g1_i2-encoding lcyE, and DN84975_c1_g7_i11-encoding CCD, which are highly related to crocin biosynthesis, were consistent with the accumulation of geniposide and crocin content, respectively. The qRT-PCR results showed that the trends of relative expression were consistent with transcribed genes. This study provides insights for understanding the geniposide and crocin accumulation and biosynthsis during fruit development in G. jasminoides. Full article

The Indo-German Science and Technology Centre (IGSTC) funded an Indo-German Workshop on Sustainable Stress Management: Aquatic plants vs. Terrestrial plants (IGW-SSMAT) which was jointly organized at the Friedrich Schiller University of Jena, Germany from 25 to 27 July 2022 by Prof. Dr. Ralf Oelmüller, Friedrich Schiller University of Jena, Germany as the German coordinator and Dr. K. Sowjanya Sree, Central University of Kerala, India as the Indian Coordinator. The workshop constituted researchers working in this field from both India and Germany and brought together these experts in the field of sustainable stress management for scientific discussions, brainstorming and networking. Full article

Phytopathogenic bacteria not only affect crop yield and quality but also the environment. Understanding the mechanisms involved in their survival is essential to develop new strategies to control plant disease. One such mechanism is the formation of biofilms; i.e., microbial communities within a three-dimensional structure that offers adaptive advantages, such as protection against unfavorable environmental conditions. Biofilm-producing phytopathogenic bacteria are difficult to manage. They colonize the intercellular spaces and the vascular system of the host plants and cause a wide range of symptoms such as necrosis, wilting, leaf spots, blight, soft rot, and hyperplasia. This review summarizes up-to-date information about saline and drought stress in plants (abiotic stress) and then goes on to focus on the biotic stress produced by biofilm-forming phytopathogenic bacteria, which are responsible for serious disease in many crops. Their characteristics, pathogenesis, virulence factors, systems of cellular communication, and the molecules implicated in the regulation of these processes are all covered. Full article

Alkalinity stress is a major hindrance to enhancing rice production globally due to its damaging effect on plants’ growth and development compared with salinity stress. However, understanding of the physiological and molecular mechanisms of alkalinity tolerance is limited. Therefore, a panel of indica and japonica rice genotypes was evaluated for alkalinity tolerance at the seedling stage in a genome-wide association study to identify tolerant genotypes and candidate genes. Principal component analysis revealed that traits such as alkalinity tolerance score, shoot dry weight, and shoot fresh weight had the highest contribution to variations in tolerance, while shoot Na⁺ concentration, shoot Na⁺:K⁺ ratio, and root-to-shoot ratio had moderate contributions. Phenotypic clustering and population structure analysis grouped the genotypes into five subgroups. Several salt-susceptible genotypes such as IR29, Cocodrie, and Cheniere placed in the highly tolerant cluster suggesting different underlying tolerance mechanisms for salinity and alkalinity tolerance. Twenty-nine significant SNPs associated with alkalinity tolerance were identified. In addition to three alkalinity tolerance QTLs, qSNK4, qSNC9, and qSKC10, which co-localized with the earlier reported QTLs, a novel QTL, qSNC7, was identified. Six candidate genes that were differentially expressed between tolerant and susceptible genotypes were selected: LOC_Os04g50090 (Helix-loop-helix DNA-binding protein), LOC_Os08g23440 (amino acid permease family protein), LOC_Os09g32972 (MYB protein), LOC_Os08g25480 (Cytochrome P450), LOC_Os08g25390 (Bifunctional homoserine dehydrogenase), and LOC_Os09g38340 (C2H2 zinc finger protein). The genomic and genetic resources such as tolerant genotypes and candidate genes would be valuable for investigating the alkalinity tolerance mechanisms and for marker-assisted pyramiding of the favorable alleles for improving alkalinity tolerance at the seedling stage in rice. Full article

Woody canker diseases caused by fungi of the Botryosphaeriaceae family are producing increasing losses in many economically important woody crops, including almond. To develop a molecular tool for the detection and quantification of the most aggressive and threatening species is of main importance. This will help to prevent the introduction of these pathogens in new orchards and to conveniently apply the appropriate control measures. Three reliable, sensitive and specific duplex qPCR assays using TaqMan probes have been designed for the detection and quantification of (a) Neofusicoccum parvum and the Neofusicoccum genus, (b) N. parvum and the Botryosphaeriaceae family and (c) Botryosphaeria dothidea and the Botryosphaeriaceae family. The multiplex qPCR protocols have been validated on artificially and naturally infected plants. Direct systems to process plant materials, without DNA purification, allowed high-throughput detection of Botryosphaeriaceae targets even in asymptomatic tissues. These results validate the qPCR using the direct sample preparation method as a valuable tool for Botryosphaeria dieback diagnosis allowing a large-scale analysis and the preventive detection of latent infection. Full article

Flower breeders are continually refining their methods for producing high-quality flowers. Phalaenopsis species are considered the most important commercially grown orchids. Advances in genetic engineering technology have provided researchers with new tools that can be used along with traditional breeding methods to enhance floral traits and quality. However, the application of molecular techniques for the breeding of new Phalaenopsis species has been relatively rare. In this study, we constructed recombinant plasmids carrying flower color-related genes, Phalaenopsis Chalcone synthase (PhCHS5) and/or Flavonoid 3′,5′-hydroxylase (PhF3′5′H). These genes were transformed into both Petunia and Phalaenopsis plants using a gene gun or an Agrobacterium tumefaciens-based method. Compared with WT, 35S::PhCHS5 and 35S::PhF3′5′H both had deeper color and higher anthocyanin content in Petunia plants. Additionally, a phenotypic comparison with wild-type controls indicated the PhCHS5 or PhF3′5′H-transgenic Phalaenopsis produced more branches, petals, and labial petals. Moreover, PhCHS5 or PhF3′5′H-transgenic Phalaenopsis both showed deepened lip color, compared with the control. However, the intensity of the coloration of the Phalaenopsis lips decreased when protocorms were co-transformed with both PhCHS5 and PhF3′5′H. The results of this study confirm that PhCHS5 and PhF3′5′H affect flower color in Phalaenopsis and may be relevant for the breeding of new orchid varieties with desirable flowering traits. Full article

Ruta chalepensis is an herb used to treat various ailments, and its potential cytotoxic effects on different tumor cell lines have been extensively studied. The present study aimed to evaluate the cytotoxic activity of R. chalepensis methanol extract (RCME), sub-partitions obtained from solvents of increasing polarity, and major compounds, as well as their hemolytic, anti-hemolytic, and antioxidant potential. The in vitro cytotoxic activity against the human hepatocarcinoma (HEP-G2) and the murine lymphoma cell line (L5178Y-R) was evaluated using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, whereas selectivity indices (SIs) were determined by comparing cytotoxicity against normal African green monkey kidney cells (VERO) and human peripheral blood mononuclear cells (PBMC). Hemolytic and anti-hemolytic activities were evaluated on human erythrocytes. The most effective cytotoxic treatment was evaluated for nitric oxide release by J774A.1 macrophages. Antioxidant activity of R. chalepensis material was also determined. Results showed that RCME produced significant (p < 0.05) cytotoxicity in HEP-G2 (IC₅₀ = 1.79 µg/mL) and L5178Y-R (IC₅₀ = 1.60 µg/mL) cells and exhibited high SIs (291.50 and 114.80, respectively). In addition, the n-hexane fraction (RCHF) showed an IC₅₀ of 18.31 µg/mL in HEP-G2 cells and an SI of 9.48 in VERO cells, whereas the chloroform fraction (RCCF) evidenced an IC₅₀ of 1.60 µg/mL in L5178Y-R cells and an SI of 34.27 in PBMC cells. Chalepensin (CHL), rutamarin (RTM), and graveolin (GRV), which are major components of R. chalepensis, showed high activity against L5178Y-R cells, with IC₅₀ of 9.15, 15.13 and SI of 45.08 µg/mL, respectively. In addition, CHL, RTM, and GRV showed SIs of 24.76, 9.98, and 3.52, respectively, when compared with PBMC cells. RCME at concentrations of 125 µg/mL and 250 µg/mL, significantly (p < 0.05) decreased nitrite production in J774A.1 cells, when exposed to lipopolysaccharide. This study demonstrated that RCME showed significant cytotoxic activity against HEP-G2 and L5178Y-R cells, without affecting normal VERO, PBMC, and J774A.1 cells. Full article

Compatibility interactions between the host and the fungal proteins are necessary to successfully establish a disease in plants by fungi or other diseases. Photochemical and antimicrobial substances are generally known to increase plant resilience, which is essential for eradicating fungus infections. Through homology modeling and in silico docking analysis, we assessed 50 phytochemicals from cucumber (Cucumis sativus), 15 antimicrobial compounds from botanical sources, and six compounds from chemical sources against two proteins of Pseudoperonospora cubensis linked to cucumber downy mildew. Alpha and beta sheets made up the 3D structures of the two protein models. According to Ramachandran plot analysis, the QNE 4 effector protein model was considered high quality because it had 86.8% of its residues in the preferred region. The results of the molecular docking analysis showed that the QNE4 and cytochrome oxidase subunit 1 proteins of P. cubensis showed good binding affinities with glucosyl flavones, terpenoids and flavonoids from phytochemicals, antimicrobial compounds from botanicals (garlic and clove), and chemically synthesized compounds, indicating the potential for antifungal activity. Full article

Plant awareness disparity (PAD, formerly plant blindness) is the human inability to notice plants in everyday life. It is suggested that the main underlying factors of PAD are: 1. the inability to recognize individual plants and 2. stronger preferences for animals, which prevents building positive attitudes toward them. The presentation of individual plants should trigger more positive responses toward them than the presentation of groups of plants. Strong preferences for animals predict that the presence of an animal on a plant might enhance positive perceptions of the plant by people. We experimentally investigated the perceived attractiveness and willingness to protect (WTP) plants presented individually and in groups and with or without various pollinators in a sample of Slovak people (N = 238). In contrast to the first prediction, only one of four plants (dog rose, but not saffron, spruce, or beech tree) received higher attractiveness scores when presented individually than in a group. None of these species received higher WTP scores when presented individually, rather than in a group. The effect of the presence of pollinators on flower attractiveness and WTP was distinguished between vertebrates and invertebrates; only flowers with birds and bats increased their attractiveness scores, while flowers with invertebrates, including a butterfly, honeybee, beetle, and the syrphid fly, received similar or lower scores than the same plant species without pollinators. WTP plants significantly increased only when the scarlet honeycreeper and the cave nectar bat were present on flowers as pollinators. People showed significantly stronger preferences for items that associate 1. plants with pollinators and 2. plants with animals that distribute animal seed than for items focused solely on plants. Connecting animals and plants should help reduce PAD. This aim cannot be achieved, however, by presenting individual plants and/or plants with randomly chosen pollinators. Full article

Solanum section Leptostemonum is an ideal lineage to test the theoretical framework regarding proposed evolutionary benefits of outcrossing sexual systems in comparison to cosexuality. Theoretically, non-cosexual taxa should support more genetic diversity within populations, experience less inbreeding, and have less genetic structure due to a restricted ability to self-fertilize. However, many confounding factors present challenges for a confident inference that inherent differences in sexual systems influence observed genetic patterns among populations. This study provides a foundational baseline of the population genetics of several species of different sexual systems with the aim of generating hypotheses of any factor—including sexual system—that influences genetic patterns. Importantly, results indicate that dioecious S. asymmetriphyllum maintains less genetic structure and greater admixture among populations than cosexual S. raphiotes at the same three locations where they co-occur. This suggests that when certain conditions are met, the evolution of dioecy may have proceeded as a means to avoid genetic consequences of self-compatibility and may support hypotheses of benefits gained through differential resource allocation partitioned across sexes. Arguably, the most significant finding of this study is that all taxa are strongly inbred, possibly reflective of a shared response to recent climate shifts, such as the increased frequency and intensity of the region’s fire regime. Full article