Project description:Drought and salinity are the major factors that limit the faba bean (Vicia faba L.) production worldwide. The aim of this study is to identify the water stress differentially expressed genes (DEGs) through the root transcriptome analyses of the drought-tolerant Hassawi 2 genotype at vegetative and flowering stages. A total of 624.8 M high-quality Illumina reads were generated and assembled into 198,155 all-unigenes with a mean length of 738 bp and an N50 length of 1347 bp. Among all-unigenes, 78,262 were assigned to non-redundant (Nr), 66,254 to nucleotide (Nt), 54,034 to KEGG, and 43,913 to gene ontology (GO) annotations. A total of 36,834 and 35,510 unigenes were differentially expressed at the vegetative and flowering stages of Hassawi 2 under drought stress, respectively. The majority of unigenes were down-regulated at both developmental stages. However, the number of genes up-regulated (15,366) at the flowering stage exceeded the number of those up-regulated (14,097) at the vegetative stage, and the number of genes down-regulated (20,144) at the flowering stage was smaller than the number of those down-regulated (22,737) at the vegetative stage. The drought stress-responsive differentially expressed unigenes coded for various regulatory proteins, including protein kinases and phosphatases, transcription factors and plant hormones and functional proteins including enzymes for osmoprotectant, detoxification and transporters were differentially expressed, most of which were largely up-regulated. Moreover, a substantial proportion of the DEGs identified in this study were novel, most exhibited a significant change in their expression levels under water stress, making them an unexploited resource that might control specific responses to drought stress in the faba bean. Finally, qRT-PCR results were found almost consistent with the results of next-generation sequencing. Our data will help in understanding the drought tolerance mechanisms in plants and will provide resources for functional genomics.
Project description:This study was carried out to identify drought-responsive genes in a drought tolerant faba bean variety (Hassawi 2) using a suppressive subtraction hybridization approach (SSH). A total of 913 differentially expressed clones were sequenced from a differential cDNA library that resulted in a total of 225 differentially expressed ESTs. The genes of mitochondrial and chloroplast origin were removed, and the remaining 137 EST sequences were submitted to the gene bank EST database (LIBEST_028448). A sequence analysis identified 35 potentially drought stress-related ESTs that regulate ion channels, kinases, and energy production and utilization and transcription factors. Quantitative PCR on Hassawi 2 genotype confirmed that more than 65% of selected drought-responsive genes were drought-related. Among these induced genes, the expression levels of eight highly up-regulated unigenes were further analyzed across 38 selected faba bean genotypes that differ in their drought tolerance levels. These unigenes included ribulose 1,5-bisphosphate carboxylase (rbcL) gene, non-LTR retroelement reverse related, probable cyclic nucleotide-gated ion channel, polyubiquitin, potassium channel, calcium-dependent protein kinase and putative respiratory burst oxidase-like protein C and a novel unigene. The expression patterns of these unigenes were variable across 38 genotypes however, it was found to be very high in tolerant genotype. The up-regulation of these unigenes in majority of tolerant genotypes suggests their possible role in drought tolerance. The identification of possible drought responsive candidate genes in Vicia faba reported here is an important step toward the development of drought-tolerant genotypes that can cope with arid environments.
Project description:Drought is a major limiting constraint to faba bean production worldwide, including Tunisia. However, molecular mechanisms underlying faba bean responses to drought stress are not well understood. In this context, transcriptome analysis by RNA-seq was performed to investigate drought-related genes and construct a network of faba bean drought stress response and tolerance. De novo assembly of the transcriptome generated 26,728 differentially expressed genes (DEGs). Of these, 13,920 were up-regulated and 12,808 down-regulated in faba bean drought-stressed leaves. Moreover, a total of 10,800 simple sequence repeats (SSRs) and 2130 transcription factors involved in major metabolic pathways including abscisic acid (ABA)-dependent and -independent signaling pathway were identified. GO, KOG and KEGG enrichment analyses revealed that these DEGs were involved in several important processes including photosynthesis, flavonoid biosynthesis, response to stimulus and abiotic stress, reactive oxygen species (ROS) scavengers, signal transduction, biosynthesis of secondary metabolites and transporters, suggesting the involvement of these important pathways in faba bean response to water deficit. Various stress proteins such as late embryogenesis abundant proteins (LEA), dehydrins (DHNs) and heat shock proteins (HSPs) have been identified and their expression was robustly upregulated in drought-stressed leaves, indicating their key contribution to drought response and adaptation by conferring protection and providing stability to faba bean plant cellular processes under water deficit. The reliability of the RNA-seq results was confirmed by the analysis of 10 randomly selected genes using qRT-PCR. Taken together, these findings help advancing our knowledge and can guide breeding programs aimed at improving the tolerance of faba bean to drought stress.
Project description:Drought stress is one of the most prevalent environmental factors limiting faba bean (Vicia faba L.) crop productivity. β-aminobutyric acid (BABA) is a non-protein amino acid that may be involved in the regulation of plant adaptation to drought stress. The effect of exogenous BABA application on physiological, biochemical and molecular responses of faba bean plants grown under 18% PEG-induced drought stress were investigated. The results showed that the application of 1 mM of BABA improved the drought tolerance of faba bean. The application of BABA increased the leaf relative water content, leaf photosynthesis rate (A), transpiration rate (E), and stomatal conductance (gs), thereby decreased the water use efficiency. Furthermore, exogenous application of BABA decreased production of hydrogen peroxide (H2O2), malondialdehyde and electrolyte leakage levels, leading to less cell membrane damage due to oxidative stress. Regarding osmoprotectants, BABA application enhanced the accumulation of proline, and soluble sugars, which could improve the osmotic adjustment ability of faba bean under drought challenge. Interestingly, mended antioxidant enzyme activities like catalase, guaiacol peroxidase, ascorbate peroxidase and superoxide dismutase and their transcript levels may lead to counteract the damaging effects of oxidative stress and reducing the accumulation of harmful substances in BABA-treated faba bean plants. In addition, exogenous BABA significantly induced the accumulation of drought tolerance-related genes like VfMYB, VfDHN, VfLEA, VfERF, VfNCED, VfWRKY, VfHSP and VfNAC in leaves and roots, suggesting that BABA might act as a signal molecule to regulate the expression of drought tolerance-related genes.
Project description:Rhizobium-driven symbiotic nitrogen-fixation in legumes not only benefits the growth but also enhances the stress tolerance of plants. Isolating and characterizing efficient, drought-tolerant rhizobia is a central goal for improving crop yields in arid regions. Here, we phylogenetically and biochemically characterized a novel strain of Rhizobium ('QHCD11') sampled from the root nodules of faba beans growing in an arid agricultural area in Qinghai-Tibet. We further tested the drought tolerance of the strain as well as of 'Qingcan 14' faba bean seedlings inoculated with it. Biochemical characterization involved bromothymol blue (BTB) tests, carbon metabolic profiling (Biolog GENIII), DNA-DNA hybridization (dDDH) assays, average nucleotide identity (ANI) analyses, and 16S rRNA sequencing. The result indicated that strain 'QHCD11' likely belongs to the Rhizobium indicum species. Drought stress tolerance was assessed by exposure to polyethylene glycol (PEG-6000) at concentrations of 0, 10, 15, and 20%. Increasing concentrations of PEG-6000 tended to result in decreased growth of 'QHCD11', although the strain performed better at 20% PEG 6000 than at 15%. Inoculation of drought-stressed faba bean seedlings with strain 'QHCD11' improved root vitality, chlorophyll content, antioxidant enzyme activities, and plant height. We suggest that inoculation of faba beans with 'QHCD11' is an environmentally sound strategy for mitigating crop drought stress in arid and semi-arid regions. In addition, the results presents here will benefit future studies into faba bean-rhizobia symbioses under drought stress.
Project description:Heat stress adversely affects the growth and yield of faba bean crop. Accumulation of ClpB/Hsp100 class of proteins is a critical parameter in induction of acquired heat stress tolerance in plants. Heat-induced expression of ClpB/Hsp100 genes has been noted in diverse plant species. Using primers complementary to soybean ClpB/Hsp100 gene, we analyzed the transcript expression profile of faba bean ClpB/Hsp100 gene in leaves of seedlings and flowering plants and in pollen grains. ClpB/Hsp100 protein accumulation profile was analyzed in leaves of faba bean seedlings using Arabidopsis thaliana cytoplasmic Hsp101 antibodies. The transcript and protein levels of faba bean ClpB/Hsp100 were significantly induced in response to heat stress.
Project description:Drought is an environmental stress that strongly impacts plants. It affects all stages of growth and induces profound disturbances that influence all cellular functions. Legumes can establish a symbiosis with Rhizobium-type bacteria, whose function is to fix atmospheric nitrogen in organs called nodules and to meet plant nitrogen needs. Symbiotic nitrogen fixation (SNF) is particularly sensitive to drought. We raised the hypothesis that, in drought-stressed nodules, SNF inhibition is partly correlated to hypoxia resulting from nodule structure compaction and an increased O2 diffusion barrier, and that the nodule energy regeneration involves phytoglobin-nitric oxide (Pgb-NO) respiration. To test this hypothesis, we subjected faba bean (Vicia faba L.) plants nodulated with a Rhizobium laguerreae strain to either drought or osmotic stress. We monitored the N2-fixation activity, the energy state (ATP/ADP ratio), the expression of hypoxia marker genes (alcohol dehydrogenase and alanine aminotransferase), and the functioning of the Pgb-NO respiration in the nodules. The collected data confirmed our hypothesis and showed that (1) drought-stressed nodules were subject to more intense hypoxia than control nodules and (2) NO production increased and contributed via Pgb-NO respiration to the maintenance of the energy state of drought-stressed nodules.
Project description:Faba bean (Vicia faba L.) is an important high protein legume adapted to diverse climatic conditions with multiple benefits for the overall sustainability of the cropping systems. Plant-based protein demand is being expanded and faba bean is a good candidate to cover this need. However, the crop is very sensitive to abiotic stresses, especially drought, which severely affects faba bean yield and development worldwide. Therefore, identifying genes associated with drought stress tolerance is a major challenge in faba bean breeding. Although the faba bean response to drought stress has been widely studied, the molecular approaches to improve drought tolerance in this crop are still limited. Here we built on recent genomic advances such as the development of the first high-density SNP genotyping array, to conduct a genome-wide association study (GWAS) using thousands of genetic polymorphisms throughout the entire faba bean genome. A worldwide collection of 100 faba bean accessions was grown under control and drought conditions and 10 morphological, phenological and physiological traits were evaluated to identify single nucleotide polymorphism (SNP) markers associated with drought tolerance. We identified 29 SNP markers significantly correlated with these traits under drought stress conditions. The flanking sequences were blasted to the Medicago truncatula reference genomes in order to annotate potential candidate genes underlying the causal variants. Three of the SNPs for chlorophyll content after the stress, correspond to uncharacterized proteins indicating the presence of novel genes associated with drought tolerance in faba bean. The significance of stress-inducible signal transducers provides valuable information on the possible mechanisms underlying the faba bean response to drought stress, thus providing a foundation for future marker-assisted breeding in the crop.
Project description:Water deficit has a significant impact on growth, development and yield of fava bean (Vicia fava L.) in arid and semi-arid climates. The aim of this study was to identify differentially expressed genes in the Qinghai 13 genotype under soil drought through leaf transcriptome analysis. A total of 256.95 M clean reads were obtained and assembled into 176334 unigenes, with an average length of 766 bp. A total of 9126 (4439 upregulated and 4687 downregulated) differentially expressed genes (DEGs) were identified in faba bean leaves under soil drought. In total, 324 putative transcription factors were identified and classified as belonging to different transcription factor families. According to GO and KEGG analysis, the soil drought stress-inducible DEGs encoded proteins mainly involved in regulating photosynthesis, osmotic adjustment, detoxification, autophagy and other functions. In addition, a large portion of DEGs appeared to be novel because they could not be annotated in any functional databases, therefore, suggesting a specific response to soil drought in faba bean. Finally, RNA-seq analysis was validated by quantitative reverse-transcription PCR analysis. This work provides comprehensive and valuable information for understanding the molecular mechanisms which faba bean uses to respond to soil drought.
Project description:Identification and validation of biomarkers and bioindicators to select genotypes with superior tolerance to water deficit (WD) under field conditions are paramount to plant breeding programs. However, the co-occurrence of different abiotic stresses such as WD, heat, and radiation makes it difficult to develop generalized protocols to monitor the physiological health of the plant system. The study assessed the most abundant carbohydrates and sugar alcohols in five faba bean (Vicia faba) genotypes under field conditions and the abundance of naturally occurring carbon isotopes in bulk leaf material to predict water use efficiency (WUE). Plant water status and biomass accumulation were also assessed. Among the accumulated sugars, inter-specific variation in glucose was most prevalent and was found at a higher concentration (8.52 mg g-1 leaf) in rainfed trial. myo-Inositol concentrations followed that of glucose accumulation in that the rainfed trial had higher amounts compared to the irrigated trial. WUE calculated from carbon isotope abundance was consistently offset with measured WUE from measurements of leaf gas exchange. All genotypes demonstrated significant relationships between predicted and measured WUE (p < 0.05) apart from control variety PBA Warda. Thus, bulk leaf-level carbon isotope abundance can be used to calculate WUE and used as an effective selection criterion for improving WUE in faba bean breeding programs under field conditions.