Epigenetic regulation influenced by soil microbiota and nutrients: Paving road to epigenome editing in plants.

Soil is a complex ecosystem that houses microbes and nutrients that are necessary for plant development. Edaphic properties of the soil and environmental conditions influence microbial growth and nutrient accessibility. Various environmental stimuli largely affect the soil microbes and ionic balance, in turn influencing plants. Soil microflora helps decompose organic matter and is involved in mineral uptake. The combination of soil microbes and mineral nutrients notably affects plant growth. Recent advancements have enabled a deeper understanding of plant genetic/molecular regulators. Deficiencies/sufficiencies of soil minerals and microbes also alter plant gene regulation. Gene regulation mediated by epigenetic mechanisms comprises conformational alterations in chromatin structure, DNA/histone modifications, or involvement of small RNAs. Epigenetic regulation is unique due to its potential to inherit without involving alteration of the DNA sequence. Thus, the compilation study of heritable epigenetic changes driven by nutrient imbalances and soil microbes would facilitate understanding this molecular phenomenon in plants. This information can aid in epigenome editing, which has recently emerged as a promising technology for plant non-transgenic/non-mutagenic modification. Potential epigenetic marks induced by biotic and abiotic stresses in plants could be explored as target sites for epigenome editing. This review discusses novel ways of epigenome editing to create epigenome edited plants with desirable and heritable phenotypes. As plants are sessile and in constant exposure to the soil microbiome and nutrients, epigenetic changes induced by these factors could provide more effective, stable and a sustainable molecular solution for crop improvement.

Candidate effectors for leaf rust resistance gene Lr28 identified through transcriptome and in-silico analysis.

Puccinia spp. causing rust diseases in wheat and other cereals secrete several specialized effector proteins into host cells. Characterization of these proteins and their interaction with host's R proteins could greatly help to limit crop losses due to diseases. Prediction of effector proteins by combining the transcriptome analysis and multiple in-silico approaches is gaining importance in revealing the pathogenic mechanism. The present study involved identification of 13 Puccinia triticina (Pt) coding sequences (CDSs), through transcriptome analysis, that were differentially expressed during wheat-leaf rust interaction; and prediction of their effector like features using different in-silico tools. NCBI-BLAST and pathogen-host interaction BLAST (PHI-BLAST) tools were used to annotate and classify these sequences based on their most closely matched counterpart in both the databases. Homology between CDSs and the annotated sequences in the NCBI database ranged from 79 to 94% and with putative effectors of other plant pathogens in PHI-BLAST from 24.46 to 54.35%. Nine of the 13 CDSs had effector-like features according to EffectorP 3.0 (≥0.546 probability of these sequences to be effector). The qRT-PCR expression analysis revealed that the relative expression of all CDSs in compatible interaction (HD2329) was maximum at 11 days post inoculation (dpi) and that in incompatible interactions (HD2329 + Lr28) was maximum at 3 dpi in seven and 9 dpi in five CDSs. These results suggest that six CDSs (>0.8 effector probability as per EffectorP 3.0) could be considered as putative Pt effectors. The molecular docking and MD simulation analysis of these six CDSs suggested that candidate Lr28 protein binds more strongly to candidate effector c14094_g1_i1 to form more stable complex than the remaining five. Further functional characterization of these six candidate effectors should prove useful for a better understanding of wheat-leaf rust interaction. In turn, this should facilitate effector-based leaf rust resistance breeding in wheat.

Brassinosteroids as promoters of seedling growth and antioxidant activity under heavy metal zinc stress in mung bean (Vigna radiata L.).

The escalation of harmful pollutants, including heavy metals, due to industrialization and urbanization has become a global concern. To mitigate the negative impacts of heavy metal stress on germination and early plant development, growth regulators have been employed. This study aimed to evaluate the response of mung bean (Vigna radiata L.) to zinc stress in the presence of brassinosteroids, focusing on seedling growth and antioxidant potential. Mung bean seedlings were treated with three concentrations of 24-epibrassinolide (EBL) (0.1, 0.2, and 0.4 PPM) with or without zinc. Results demonstrated that the application of brassinosteroids, combined with zinc stress, significantly enhanced germination percentage (about 47.06, 63.64, and 120%), speed of germination (about 39.13, 50, and 100%), seedling growth (about 38% in case of treatment combined 0.4 PPM 24-EBL and 1.5 mM ZnSO4) and seedling vigor index (204% in case of treatment combined 0.4 PPM 24-EBL and 1.5 mM ZnSO4) compared to zinc-treated seedlings alone after 24 h. The activities of antioxidative enzymes (catalase, ascorbate peroxidase, polyphenol oxidase, and peroxidase) and total soluble protein content decreased, while lipid peroxidation and proline content exhibited a significant increase (p ≤ 0.05) when compared to the control. However, the negative effects induced by heavy metal stress on these parameters were significantly mitigated by EBL application. Notably, the most effective concentration of EBL in overcoming zinc stress was found to be 0.4 PPM. These findings underscore the potential of exogenously applied brassinosteroids as a valuable tool in phytoremediation projects by ameliorating heavy metal stress.

Virulence and genetic analysis of Puccinia graminis tritici in the Indian sub-continent from 2016 to 2022 and evaluation of wheat varieties for stem rust resistance.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), has re-emerged as one of the major concerns for global wheat production since the evolution of Ug99 and other virulent pathotypes of Pgt from East Africa, Europe, Central Asia, and other regions. Host resistance is the most effective, economic, and eco-friendly approach for managing stem rust. Understanding the virulence nature, genetic diversity, origin, distribution, and evolutionary pattern of Pgt pathotypes over time and space is a prerequisite for effectively managing newly emerging Pgt isolates through host resistance. In the present study, we monitored the occurrence of stem rust of wheat in India and neighboring countries from 2016 to 2022, collected 620 single-pustule isolates of Pgt from six states of India and Nepal, analyzed them on Indian stem rust differentials, and determined their virulence phenotypes and molecular genotypes. The Ug99 type of pathotypes did not occur in India. Pathotypes 11 and 40A were most predominant during these years. Virulence phenotyping of these isolates identified 14 Pgt pathotypes, which were genotyped using 37 Puccinia spp.-specific polymorphic microsatellites, followed by additional phylogenetic analyses using DARwin. These analyses identified three major molecular groups, demonstrating fewer lineages, clonality, and long-distance migration of Pgt isolates in India. Fourteen of the 40 recently released Indian wheat varieties exhibited complete resistance to all 23 Pgt pathotypes at the seedling stage. Twelve Sr genes were postulated in 39 varieties based on their seedling response to Pgt pathotypes. The values of slow rusting parameters i.e. coefficient of infection, area under disease progress curve, and infection rates, assessed at adult plant stage at five geographically different locations during two crop seasons, indicated the slow rusting behavior of several varieties. Six Sr genes (Sr2, Sr57, Sr58, Sr24, Sr31, and Sr38) were identified in 24 wheat varieties using molecular markers closely linked to these genes. These findings will guide future breeding programs toward more effective management of wheat stem rust.

Dataset on antioxidant system of non-model halophytes Urochondra setulosa and Dichanthium annulatum in saline environment.

The antioxidant potential of halophytes, Dichanthium annulatum and Urochondra setulosa, was examined under the influence of high salinity. These halophytes were grown in lysimeters filled with saline soil and further irrigated with saline water to maintain different salt levels of ECe 30, 40 and 50 dS m-1 along with the one set in normal field soil without saline irrigation serving as control. The leaf samples were collected after saline irrigation and analyzed for the antioxidative enzymes i.e., Catalase (CAT), Peroxidase (POX), Superoxide dismutase (SOD), Ascorbate peroxidase (APX), Monodehydroascorbate reductase (MDHAR), Dehydroascorbate reductase (DHAR) and Glutathione reductase (GR), including the ROS metabolites such as H2O2 content, malondialdehyde content (MDA), ascorbic acid content and total glutathione content. The mechanism of scavenging the reactive oxygen species in both the halophytes was characterized.

Unraveling the diversity and functions of sugar transporters for sustainable management of wheat rust.

Plant diseases threaten global food security by reducing the production and quality of produce. Identification of disease resistance sources and their utilization in crop improvement is of paramount significance. However, constant evolution and occurrence of new, more aggressive and highly virulent pathotypes disintegrates the resistance of cultivars and hence demanding the steady stream of disease resistance cultivars as the most sustainable way of disease management. In this context, molecular tools and technologies facilitate an efficient and rational engineering of crops to develop cultivars having resistance to multiple pathogens and pathotypes. Puccinia spp. is biotrophic fungi that interrupt crucial junctions for causing infection, thus risking nutrient access of wheat plants and their subsequent growth. Sugar is a major carbon source taken from host cells by pathogens. Sugar transporters (STPs) are key players during wheat-rust interactions that regulate the transport, exchange, and allocation of sugar at plant-pathogen interfaces. Intense competition for accessing sugars decides fate of incompatibility or compatibility between host and the pathogen. The mechanism of transport, allocation, and signaling of sugar molecules and role of STPs and their regulatory switches in determining resistance/susceptibility to rusts in wheat is poorly understood. This review discusses the molecular mechanisms involving STPs in distribution of sugar molecules for determination of rust resistance/susceptibility in wheat. We also present perspective on how detailed insights on the STP's role in wheat-rust interaction will be helpful in devising efficient strategies for wheat rust management.

Halophytes as new model plant species for salt tolerance strategies.

Soil salinity is becoming a growing issue nowadays, severely affecting the world's most productive agricultural landscapes. With intersecting and competitive challenges of shrinking agricultural lands and increasing demand for food, there is an emerging need to build resilience for adaptation to anticipated climate change and land degradation. This necessitates the deep decoding of a gene pool of crop plant wild relatives which can be accomplished through salt-tolerant species, such as halophytes, in order to reveal the underlying regulatory mechanisms. Halophytes are generally defined as plants able to survive and complete their life cycle in highly saline environments of at least 200-500 mM of salt solution. The primary criterion for identifying salt-tolerant grasses (STGs) includes the presence of salt glands on the leaf surface and the Na+ exclusion mechanism since the interaction and replacement of Na+ and K+ greatly determines the survivability of STGs in saline environments. During the last decades or so, various salt-tolerant grasses/halophytes have been explored for the mining of salt-tolerant genes and testing their efficacy to improve the limit of salt tolerance in crop plants. Still, the utility of halophytes is limited due to the non-availability of any model halophytic plant system as well as the lack of complete genomic information. To date, although Arabidopsis (Arabidopsis thaliana) and salt cress (Thellungiella halophila) are being used as model plants in most salt tolerance studies, these plants are short-lived and can tolerate salinity for a shorter duration only. Thus, identifying the unique genes for salt tolerance pathways in halophytes and their introgression in a related cereal genome for better tolerance to salinity is the need of the hour. Modern technologies including RNA sequencing and genome-wide mapping along with advanced bioinformatics programs have advanced the decoding of the whole genetic information of plants and the development of probable algorithms to correlate stress tolerance limit and yield potential. Hence, this article has been compiled to explore the naturally occurring halophytes as potential model plant species for abiotic stress tolerance and to further breed crop plants to enhance salt tolerance through genomic and molecular tools.

Genetic Diversity and Population Structure Reveal Cryptic Genetic Variation and Long Distance Migration of Puccinia graminis f. sp. tritici in the Indian Subcontinent.

Stem rust caused by Puccinia graminis f. sp. tritici (Pgt) is a devastating disease of wheat worldwide since time immemorial. Several wheat stem rust outbreaks have been reported worldwide including India. Approximately 7 mha wheat area in central and peninsular India is highly vulnerable to stem rust epidemics. In this study, a repository of 29 single genotype uredospore pathotypes, representing five geographical regions, was characterized by investigating their virulence phenotype and simple sequence repeat (SSR) genotypes using 37 reproducible polymorphic SSR markers, 32 of which had ≥ 0.50 polymorphic information content (PIC) value. Virulence phenotypes were used to evaluate the virulence frequency (VF) and construct a hypothetical evolutionary hierarchy of these pathotypes. We projected seven lineages to explain the evolutionary pattern of the Pgt population. The VF of these pathotypes ranged between 0% and 100%. The virulence-based neighbor-joining (NJ) cluster analysis grouped Pgt pathotypes into five virulence groups. Likewise, five molecular groups were categorized using molecular genotypes. The molecular grouping was supported by principal coordinate analysis (PCoA), which revealed 25% of the cumulative variance contributed by the first two axes. Analysis of molecular variance (AMOVA) revealed 8 and 92% of the variation among and within the populations, respectively. The Mantel test confirmed a positive but weak correlation (R 2 = 0.15) between virulence phenotypes and SSR genotypes. The highest and lowest values of different genetic diversity parameters (Na, Ne, I, He, uHe, and %P) revealed maximum and minimum variability in the Pgt population from Maharashtra and Uttar Pradesh, respectively. The population structure analysis clustered 29 Pgt pathotypes into two subpopulations and an admixture. Our results demonstrated that there was significant genetic diversity among Pgt pathotypes resulting from their long-distance dispersal ability complemented by gene flow. These findings provide insights into the virulence patterns, genetic variations, and possible evolution of Pgt pathotypes, which would support strategic stem rust resistance breeding.

Elucidating the Population Structure and Genetic Diversity of Indian Puccinia striiformis f. sp. tritici Pathotypes Based on Microsatellite Markers.

In India, systematic wheat yellow rust survey and pathotype (race) analysis work began in 1930. However, information on population structure and genetic diversity of yellow rust pathogen has not been available. To address this, we conducted studies on population structure and genetic diversity of Puccinia striiformis f. sp. tritici (Pst) pathotypes using 38 simple sequence repeat primer-pairs. Bayesian assignment and discriminant analysis of principal components indicated the presence of two distinct Pst subpopulations (Pop1 and Pop2) along with 37.9% admixed pathotypes. The unweighted pair-group method with arithmetic mean also categorized these pathotypes into two major clusters. Principal coordinates analysis explained 20.06 and 12.50% variance in horizontal and vertical coordinates, respectively. Index of association (IA) and the standardized index of association ([Formula: see text]) values showed that Pst subpopulations reproduced asexually (clonally). In total, 102 alleles were detected, with the expected heterozygosity (Hexp) per locus ranging from 0.13 to 0.73, with a mean of 0.47. The average polymorphic information content value of 0.40 indicated high genetic diversity among pathotypes. Analysis of molecular variance revealed 12% of the total variance between subpopulations, 11% among the pathotypes of each subpopulation, and 77% within pathotypes. A significant moderate level of genetic differentiation (FST = 0.122, P < 0.001) and gene flow (Nm = 1.80) were observed between subpopulations. The Pst virulence phenotypes showed a weak positive correlation (R2 = 0.027, P < 0.02) with molecular genotypes.

microRNA 166: an evolutionarily conserved stress biomarker in land plants targeting HD-ZIP family.

MicroRNAs (miRNAs) are significant class of noncoding RNAs having analytical investigating and modulatory roles in various signaling mechanisms in plants related to growth, development and environmental stress. Conserved miRNAs are an affirmation of land plants evolution and adaptation. They are a proof of indispensable roles of endogenous gene modulators that mediate plant survival on land. Out of such conserved miRNA families, is one core miRNA known as miR166 that is highly conserved among land plants. This particular miRNA is known to primarily target HD ZIP-III transcription factors. miR166 has roles in various developmental processes, as well as regulatory roles against biotic and abiotic stresses in major crop plants. Major developmental roles indirectly modulated by miR166 include shoot apical meristem and vascular differentiation, leaf and root development. In terms of abiotic stress, it has decisive regulatory roles under drought, salinity, and temperature along with biotic stress management. miR166 and its target genes are also known for their beneficial synergy with microorganisms in leguminous crops in relation to lateral roots and nodule development. Hence it is important to study the roles of miR166 in different crop plants to understand its defensive roles against environmental stresses and improve plant productivity by reprogramming several gene functions at molecular levels. This review is hence a summary of different regulatory roles of miR166 with its target HD-ZIP III and its modulatory and fine tuning against different environmental stresses in various plants.

De novo transcriptomic data of salt tolerant halophytes Dichnathium annulatum (Forssk.) stapf and Urochondra setulosa (Trin.) C.E.Hubb.

Two halophytes, Dichanthium annulatum (moderately salt tolerant) and Urochondra setulosa (highly salt tolerant) were selected to generate transcriptome at different salinity levels. Sequencing of RNA samples was done on Illumina-Hi-Seq platform for de novo transcriptome assembly from the leaf tissues of D. annulatum at salinity of ECe ∼30 dS/m and of U. setulosa at three salt levels (i.e. ECe ∼30, ∼40 and ∼50 dS/m). DESeq was used for identification of differentially expressed transcripts and a total of 267,196 and 384,442 transcripts were assembled through Trinity in both the plants respectively. A total of 32,246 and 25,479 SSRs were identified respectively in both the plants using MISA perl script with mono and tri-nucleotide repeats as most common motif.

Role of nanoparticles in crop improvement and abiotic stress management.

Nanoparticles (NPs) possess specific physical and chemical features and they are capable enough to cross cellular barriers and show their effect on living organisms. Their capability to cross cellular barriers have been noticed for their application not only in medicine, electronics, chemical and physical sciences, but also in agriculture. In agriculture, nanotechnology can help to improve the growth and crop productivity by the use of various nanoscale products such as nanofertilizers, nanoherbicides, nanofungicides, nanopesticides etc. An optimized concentration of NPs can be administered by incubation of seeds, roots, pollen, isolated cells and protoplast, foliar spraying, irrigation with NPs, direct injection, hydroponic treatment and delivery by biolistics. Once NPs come in contact with plant cells, they are uptaken by plasmodesmatal or endocytosed pathways and translocated via apoplastic and / symplastic routes. Once beneficial NPs reach different parts of plants, they boost photosynthetic rate, biomass measure, chlorophyll content, sugar level, buildup of osmolytes and antioxidants. NPs also improve nitrogen metabolism, enhance chlorophyll as well as protein content and upregulate the expression of abiotic- and biotic stress-related genes. Herein, we review the state of art of different modes of application, uptake, transport and prospective beneficial role of NPs in stress management and crop improvement.

An overview on miRNA-encoded peptides in plant biology research.

MicroRNAs (miRNAs) are short (21-23 nt) regulatory RNA molecules present in plants and animals which are known for regulating the mRNA target gene expression either by cleavage or translational repression. With the advancements in miRNAs research in plants towards their biogenesis and applications has directed the recent discovery of pri-miRNAs encoding functional peptides or microRNA peptides (miPEPs). These miPEPs are encoded by 5' of pri-miRs containing short ORFs (miORFs). miPEPs are known to enhance the activity of their associated miRNAs by increasing their accumulation and hence downregulating the target genes. Since miPEPs are very specific for each miRNA, they are considered as novel and effective tools for improving traits of interest for plant growth promotion and plant-microbe interaction. Entire peptidome research is the need of the hour. This review thus summarizes recent advancements in miPEP research and its applications as a technology with important agronomical implications with miRNAs augmentation.

Reference gene identification for gene expression analysis in rice under different metal stress.

Real-time quantitative polymerase chain reaction (RT-qPCR) is the most common approach to quantify changes in gene expression. Appropriate internal reference genes are essential for normalization of data of RT-qPCR. In the present study, we identified suitable reference genes for analysis of gene expression in rice seedlings subjected to different heavy metal stresses such as deficiencies of iron and zinc and toxicities of cobalt, cadmium and nickel. First, from publically available RNA-Seq data we identified 10 candidate genes having stable expression. We also included commonly used house-keeping gene OsUBQ5 (Ubiquitin 5) in our analysis. Expression stability of all the 11 genes was determined by two independent tools, NormFinder and geNorm. Our results show that selected candidate reference genes have higher stability in their expression compared to that of OsUBQ5. Genes with locus ID LOC_Os03g16690, encoding an oxysterol-binding protein (OsOBP) and LOC_Os01g56580, encoding Casein Kinase_1a.3 (OsCK1a.3) were identified to be the most stably expressed reference genes under most of the conditions tested. Finally, the study reveals that it is better to use a specific reference gene for a specific heavy metal stress condition rather than using a common reference gene for multiple heavy metal stress conditions. The reference genes identified here would be very useful for gene expression studies under heavy metal stresses in rice.

Proteomic approach to identify the differentially abundant proteins during flavour development in tuberous roots of Decalepis hamiltonii Wight & Arn.

2-Hydroxy-4-Methoxy Benzaldehyde (2H4MB) is a structural isomer of vanillin produced in the tuberous roots of D. hamiltonii. Both vanillin and 2H4MB share the common phenylpropanoid pathway for their synthesis. Unlike vanillin, in which the biosynthetic pathway was well elucidated in V. planifolia, the 2H4MB biosynthetic pathway is not known in any of its plant sources. To find the key enzymes/proteins that promote 2H4MB biosynthesis, a comparative proteomic approach was adapted. In this case, two developmental stages of tuberous roots of D. hamiltonii were selected, where the flavour content was highly variable. The flavour content in the two stages was estimated using quantitative HPLC. The flavour content in the first and second stages of tuber development was 160 and 510 µgg-1, respectively. Two-dimensional electrophoresis (2-DE) was performed for these two stages of tubers; this was followed by PDquest analysis. A total of 180 protein spots were differentially abundant of which 57 spots were selected and subjected to MALDI-TOF-TOF analysis. The largest percentage of identified proteins was involved in stress and defence (27.9%), followed by proteins related to bioenergy and metabolism (23.2%), Cellular homeostasis proteins (18.6%), signaling proteins (11.6%), Plant growth and development proteins (9.3%). Holistically, we found the upregulation of methyltransferase, cell division responsive proteins, plant growth and development proteins which directly relate to flavour development and maturation. Similarly, stress-responsive and signaling proteins, vacuole proteins and ATPases were down-regulated with an increase in flavour content. In this study, we could not identify the specific 2H4MB metabolic pathway proteins, however, we could be able to study the changes in physiological and primary metabolic proteins with 2H4MB accumulation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02714-x.

Effect of saline irrigation on plant water traits, photosynthesis and ionic balance in durum wheat genotypes.

In the era of climate change, decreased precipitation and increased evapo-transpiration hampers the yield of several cereal crops along with the soil salinity and poor ground water resource. Wheat being the moderately tolerant crop face many challenges in the arid and semi-arid regions under irrigated agriculture. In view of this, the study was planned to explore the potential of durum wheat genotypes under salinity on the basis of physiological traits. Experiment was designed as RBD in three replications to evaluate 15 wheat genotypes with moderate saline irrigation (ECiw - 6 dS m-1) and extreme saline irrigation (ECiw - 10 dS m-1) along with one set of control (Best available water). Different physiological traits such as water potential (ψp), osmotic potential (ψs), relative water content (RWC), Na+ and K+ content were recorded in roots as well as shoots at the reproductive stage whereas photosynthetic rate and chlorophyll content were measured in the flag leaves. A significant variability (p < 0.001) was noted among the genotypes under different stress environments and it was observed that durum genotype HI 8728 and HI 8737 showed less reduction in plant water traits (RWC, ψp and ψs) than the salinity tolerant checks of bread wheat KRL 99 and KRL 3-4. HD 4728 and HI 8708 maintained higher photosynthetic rate as well as higher chlorophyll content under the extreme salinity level of ECiw - 10 dSm-1. No significant differences were found in root Na+ in genotypes KRL 99 (3.17g), KRL 3-4 (3.34g) and HI 8737 (3.41g) while in shoots, lowest accumulation was seen in KRL 99, MACS 3949 and KRL 3-4 at ECiw - 10 dSm-1. The mean range of K+ content was 7.60-9.74% in roots and 4.21-6.61% in shoots under control environment which decreased to 50.77% in roots and 46.05% in shoots under extreme salinity condition of ECiw - 10 dSm-1. At ECiw - 10 dSm-1, KRL 99 maintained highest K+/Na+ in both root and shoot followed by KRL 3-4, HI 8737, MACS 3949, HD 4728 in roots and MACS 3949, KRL 3-4, MACS 4020, HD 4758, MACS 3972 and HI 8713 in shoots. The differential response of durum wheat genotypes under salinity particularly for physiological traits, confer their adaptability towards stress environments and exhibit their potential as genetic sources in breeding programs for improving salt stress tolerance.

de novo transcriptomic profiling of differentially expressed genes in grass halophyte Urochondra setulosa under high salinity.

Soil salinity is one of the major limiting factors for crop productivity across the world. Halophytes have recently been a source of attraction for exploring the survival and tolerance mechanisms at extreme saline conditions. Urochondra setulosa is one of the obligate grass halophyte that can survive in up to 1000 mM NaCl. The de novo transcriptome of Urochondra leaves at different salt concentrations of 300-500 mM NaCl was generated on Illumina HiSeq. Approximately 352.78 million high quality reads with an average contig length of 1259 bp were assembled de novo. A total of 120,231 unigenes were identified. On an average, 65% unigenes were functionally annotated to known proteins. Approximately 35% unigenes were specific to Urochondra. Differential expression revealed significant enrichment (P < 0.05) of transcription factors, transporters and metabolites suggesting the transcriptional regulation of ion homeostasis and signalling at high salt concentrations in this grass. Also, about 143 unigenes were biologically related to salt stress responsive genes. Randomly selected genes of important pathways were validated for functional characterization. This study provides useful information to understand the gene regulation at extremely saline levels. The study offers the first comprehensive evaluation of Urochondra setulosa leaf transcriptome. Examining non-model organisms that can survive in harsh environment can provide novel insights into the stress coping mechanisms which can be useful to develop improved agricultural crops.

An OsNAM gene plays important role in root rhizobacteria interaction in transgenic Arabidopsis through abiotic stress and phytohormone crosstalk.

KEY MESSAGE: Overexpression of Bacillus amyloliquefaciens SN13-responsive OsNAM gene in Arabidopsis reveals its important role in beneficial plant and plant growth promoting rhizobacteria interaction by conferring stress tolerance and phytohormone modulation. Salinity is one of the major constraints that affect crop development and yield. Plants respond and adapt to salt stress via complex mechanisms that involve morpho-physiological, biochemical, and molecular changes. The expression of numerous genes is known to alter during various abiotic stresses and impart stress tolerance. Recently, some known rhizospheric microbes have also been used to mitigate the effects of abiotic stresses; however, the molecular basis of such interactions remains elusive. Therefore, the present investigation was aimed to elucidate the plant growth-promoting rhizobacteria (PGPR; Bacillus amyloliquefaciens-SN13) -induced crosstalk among salinity and phytohormones in OsNAM-overexpressed Arabidopsis plants. Transgenic plants showed increased germination percentage compared to wild-type (WT) seeds under 100 mM of NaCl. Phenotypic data showed increased root length, rosette diameter, leaf size, and biomass in transgenics than WT plants. Transgenic plants can also better maintain membrane integrity and osmolyte concentration under salinity as compared to WT. Further, gene expression analysis of AP2/ERF, GST, ERD4, and ARF2 genes showed differential expression and their positive modulation in transgenic Arabidopsis exposed to salt stress in the presence of SN13 as compared to uninoculated WT. Modulation in IAA, ABA, and GA content in inoculated plants showed the more pronounced positive effects of SN13 on transgenic plants that supported our findings on Arabidopsis-SN13 interaction. Overall, the study concludes that SN13 positively modulated expression of stress-responsive genes under salinity and alter phytohormones levels in OsNAM-overexpressed plants suggesting its extensive role in cross-talk among salinity and phytohormones in response to PGPR.

Genome-wide identification and expression analysis of Arabidopsis GRAM-domain containing gene family in response to abiotic stresses and PGPR treatment.

Characterization of stress-responsive genes is important to understand the genomics perspective of stress tolerance. In this purview, several gene-families are being identified and characterized in the model and non-model plant species, which has greatly enhanced the knowledge of molecular intricacies associated with stress tolerance. One such gene family is the GRAM-domain containing which have been found to be upregulated in response to plant growth-promoting rhizobacteria (PGPR) treatment followed by salinity stress. Thus, we aimed at understanding the involvement of GRAM domain-containing proteins in abiotic stress response under the influence of rhizobacteria in Arabidopsis thaliana. The study identified fourteen AtGRAM genes in A. thaliana. Further, comprehensive analyses of domain family including phylogenetic studies, domain architecture, gene structure and genomic composition analysis, promoter analysis, homology modelling, and duplication and divergence rates estimation was performed. RNA-Seq derived expression profiling of AtGRAM genes using GENVESTIGATOR in different stresses, developmental stages and hormonal treatments was performed, followed by qRT-PCR analysis under abiotic stresses in response to PGPR. Altogether, the study provided insights into the structure, organization, and evolutionary properties of AtGRAM gene family. Modulation in expression pattern in response to stresses influenced by PGPR-treatment suggests its multifaceted role in cross-talk among abiotic stresses and phytohormones. Further functional characterization of the selected candidate genes would enable understanding of the precise roles of GRAM-genes underlying stress tolerance.