Seed endophytic bacterium Lysinibacillus sp. (ZM1) from maize (Zea mays L.) shapes its root architecture through modulation of auxin biosynthesis and nitrogen metabolism.

Seed endophytic bacteria have been shown to promote the growth and development of numerous plants. However, the underlying mechanism still needs to be better understood. The present study aims to investigate the role of a seed endophytic bacterium Lysinibacillus sp. (ZM1) in promoting plant growth and shaping the root architecture of maize seedlings. The study explores how bacteria-mediated auxin biosynthesis and nitrogen metabolism affect plant growth promotion and shape the root architecture of maize seedlings. The results demonstrate that ZM1 inoculation significantly enhances root length, root biomass, and the number of seminal roots in maize seedlings. Additionally, the treated seedlings exhibit increased shoot biomass and higher levels of photosynthetic pigments. Confocal laser scanning microscopy (CLSM) analysis revealed extensive colonization of ZM1 on root hairs, as well as in the cortical and stellar regions of the root. Furthermore, LC-MS analysis demonstrated elevated auxin content in the roots of the ZM1 treated maize seedlings compared to the uninoculated control. Inoculation with ZM1 significantly increased the levels of endogenous ammonium content, GS, and GOGAT enzyme activities in the roots of treated maize seedlings compared to the control, indicating enhanced nitrogen metabolism. Furthermore, inoculation of bacteria under nitrogen-deficient conditions enhanced plant growth, as evidenced by increased root shoot length, fresh and dry weights, average number of seminal roots, and content of photosynthetic pigments. Transcript analysis indicated upregulation of auxin biosynthetic genes, along with genes involved in nitrogen metabolism at different time points in roots of ZM1-treated maize seedlings. Collectively, our findings highlight the positive impact of Lysinibacillus sp. ZM1 inoculation on maize seeds by improving root architecture through modulation of auxin biosynthesis and affecting various nitrogen metabolism related parameters. These findings provide valuable insights into the potential utilization of seed endophytic bacteria as biofertilizers to enhance plant growth and yield in nutrient deficient soils.

Metabolomic profiling reveals key factors and associated pathways regulating the differential behavior of rice (Oryza sativa L.) genotypes exposed to geogenic arsenic.

Arsenic (As) toxicity is an escalating problem; however, information about the metabolic events controlling the varied pattern of As accumulation in rice genotypes within their natural environment is still lacking. The present study is thus an advancement in unravelling the response of such rice genotypes. Soil-water-rice samples were analyzed for As accumulation using ICP-MS. Furthermore, we implemented metabolomics through LC-MS/MS and UHPLC to identify metabolic signatures regulating As content by observing the metalloid's composition in rice agrosystem. Results showed that rice genotypes differed significantly in their levels of metabolites, with Mini mansoori and Pioneer having the highest levels. Mini mansoori contained least As which might have been regulated by Ala, Ser, Glu, Phe, Asn, His, Ile, Lys, Gln, Trp, Tyr, chlorogenic, p-coumaric, trans-ferulic, rutin, morin, naringenin, kampferol, and myricetin, while Asp, Arg, Met, syringic, epigalocatechin, and apigenin contributed to the greater As acclimatization ability of Pioneer. Multivariate tools separated the rice genotypes into two major clusters: Pioneer-Mini mansoori and Damini-Sampoorna-Chintu. KEGG identified three major metabolic pathways (aminoacyl-tRNA, phenylpropanoid, and secondary metabolites biosynthesis route) linked with As tolerance and adaptation mechanisms in rice. Overall, these two genotypes symbolize their As hostile and accommodating attitudes probably due to the accumulated metabolites and the physicochemical attributes of the soil-water. Thus, thorough understanding of the metabolic reactions to As may facilitate the emergence of As tolerant/resilient genotypes. This will aid in the selection of molecular markers to cultivate healthier rice genotypes in As-contaminated areas.

Functional characterization of 2-oxoglutarate-dependent dioxygenase gene family in chickpea.

Chickpea is an important leguminous crop plant with two cultivated types, desi and kabuli. It is nutritionally enriched in flavonoid content in addition to minerals and vitamins imparting huge health benefits to human beings. Our study elucidates the functionality of 2-oxoglutarate dependent dioxygenase (2-ODD) gene family members i.e., flavanone-3-hydroxylase (F3H), flavonol synthase (FLS) and anthocyanidin synthase (ANS) in chickpea using heterologous bacterial system and in-planta studies in Arabidopsis. This provides information about the biosynthesis of two very significant sub-classes of flavonoids- flavonols and anthocyanins. Here, we show that all the three homologs of F3H in chickpea can utilize not just naringenin but also eriodictyol as their substrate. Moreover, we show that FLS in chickpea exhibits bifunctionality having both FLS and F3H activity. Also, our study indicates the richness of desi chickpea over kabuli type through gene expression and metabolite content analyses. Overall, our study establishes the functionality of 2-ODD gene family involved in the early and late steps of flavonoid biosynthesis pathway in chickpea. It paves way for better genetic manipulation of the pathway for direct or indirect synthesis of three major subclasses of flavonoids (flavonol, anthocyanin and proanthocyanin) to develop nutritious, environmentally stable and healthy chickpea (Cicer arietinum) crop.

The R2R3-MYB-SG7 transcription factor CaMYB39 orchestrates surface phenylpropanoid metabolism and pathogen resistance in chickpea.

Flavonoids are important plant pigments and defense compounds; understanding the transcriptional regulation of flavonoid biosynthesis may enable engineering crops with improved nutrition and stress tolerance. Here, we characterize R2R3-MYB domain subgroup 7 transcription factor CaMYB39, which regulates flavonol biosynthesis primarily in chickpea trichomes. CaMYB39 overexpression in chickpea was accompanied by a change in flux availability for the phenylpropanoid pathway, particularly flavonol biosynthesis. Lines overexpressing CaMYB39 showed higher isoflavonoid levels, suggesting its role in regulating isoflavonoid pathway. CaMYB39 transactivates the transcription of early flavonoid biosynthetic genes (EBG). FLAVONOL SYNTHASE2, an EBG, encodes an enzyme with higher substrate specificity for dihydrokaempferol than other dihydroflavonols explaining the preferential accumulation of kaempferol derivatives as prominent flavonols in chickpea. Interestingly, CaMYB39 overexpression increased trichome density and enhanced the accumulation of diverse flavonol derivatives in trichome-rich tissues. Moreover, CaMYB39 overexpression reduced reactive oxygen species levels and induced defense gene expression which aids in partially blocking the penetration efficiency of the fungal pathogen, Ascochyta rabiei, resulting in lesser symptoms, thus establishing its role against deadly Ascochyta blight (AB) disease. Overall, our study reports an instance where R2R3-MYB-SG7 member, CaMYB39, besides regulating flavonol biosynthesis, modulates diverse pathways like general phenylpropanoid, isoflavonoid, trichome density, and defense against necrotrophic fungal infection in chickpea.

The nuclear effector ArPEC25 from the necrotrophic fungus Ascochyta rabiei targets the chickpea transcription factor CaßLIM1a and negatively modulates lignin biosynthesis, increasing host susceptibility.

Fungal pathogens deploy a barrage of secreted effectors to subvert host immunity, often by evading, disrupting, or altering key components of transcription, defense signaling, and metabolic pathways. However, the underlying mechanisms of effectors and their host targets are largely unexplored in necrotrophic fungal pathogens. Here, we describe the effector protein Ascochyta rabiei PEXEL-like Effector Candidate 25 (ArPEC25), which is secreted by the necrotroph A. rabiei, the causal agent of Ascochyta blight disease in chickpea (Cicer arietinum), and is indispensable for virulence. After entering host cells, ArPEC25 localizes to the nucleus and targets the host LIM transcription factor CaßLIM1a. CaßLIM1a is a transcriptional regulator of CaPAL1, which encodes phenylalanine ammonia lyase (PAL), the regulatory, gatekeeping enzyme of the phenylpropanoid pathway. ArPEC25 inhibits the transactivation of CaßLIM1a by interfering with its DNA-binding ability, resulting in negative regulation of the phenylpropanoid pathway and decreased levels of intermediates of lignin biosynthesis, thereby suppressing lignin production. Our findings illustrate the role of fungal effectors in enhancing virulence by targeting a key defense pathway that leads to the biosynthesis of various secondary metabolites and antifungal compounds. This study provides a template for the study of less explored necrotrophic effectors and their host target functions.

Biochemical analysis of anthocyanin and proanthocyanidin and their regulation in determining chickpea flower and seed coat colour.

Flower and seed coat colour are important agronomic traits in chickpea (Cicer arietinum L.). Cultivated chickpeas are of two types namely, desi (dark seeded, purple flowered) and kabuli (light seeded, white flowered). There has been limited information about the molecular mechanism underlying colour variation of flower and seed coats in desi and kabuli chickpea. We profiled the anthocyanin and proanthocyanidin (PA) contents in chickpea flowers and seed coats. Tissue-specific silencing of two genes encoding a basic helix-loop-helix (CabHLH) protein and a tonoplast-localized multidrug and toxic compound extrusion (CaMATE1) transporter in a desi genotype resulted in the reduction in expression of anthocyanin and PA biosynthetic genes and anthocyanin and PA contents in the flower and seed coat, and produced flowers and seeds with kabuli characteristics. Transcriptional regulation of a subset of anthocyanin and PA biosynthetic genes by a natural CabHLH variant and transport assay of a natural CaMATE1 variant explained the association of these alleles with the kabuli phenotype. We carried out a detailed molecular characterization of these genes, and provided evidence that kabuli chickpea flower and seed colour phenotypes can be derived by manipulation of single genes in a desi chickpea background.

Endophytic Burkholderia: Multifunctional roles in plant growth promotion and stress tolerance.

The genus Burkholderia has proven potential in improving plant performance. In recent decades, a huge diversity of Burkholderia spp. have been reported with diverse capabilities of plant symbiosis which could be harnessed to enhance plant growth and development. Colonization of endophytic Burkholderia spp. have been extensively studied through techniques like advanced microscopy, fluorescent labelling, PCR based assays, etc., and found to be systemically distributed in plants. Thus, use of these biostimulant microbes holds the promise of improving quality and quantity of crops. The endophytic Burkholderia spp. have been found to support plant functions along with boosting nutrient availability, especially under stress. Endophytic Burkholderia spp. improve plant survival against deadly pathogens via mechanisms like competition, induced systemic resistance, and antibiosis. At the same time, they are reported to extend plant tolerance towards multiple abiotic stresses especially drought, salinity, and cold. Several attempts have been made to decipher the potential of Burkholderia spp. by genome mining, and these bacteria have been found to harbour genes for plant symbiosis and for providing multiple benefits to host plants. Characteristics specific for host recognition and nutrient acquisition were confirmed in endophytic Burkholderia by genomics and proteomics-based studies. This could pave the way for harnessing Burkholderia spp. for biotechnological applications like biotransformation, phytoremediation, insecticidal activity, antimicrobials, etc. All these make Burkholderia spp. a promising microbial agent in improving plant performance under multiple adversities. Thus, the present review highlights critical roles of endophytic Burkholderia spp., their colonization, alleviation of biotic and abiotic stresses, biotechnological applications and genomic insights.