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Transcriptome profiling of a beach-adapted wild legume for dissecting novel mechanisms of salinity tolerance

ABSTRACT: Strophostyles helvola is a close relative to common bean (Phaseolus vulgaris) and inhabits both coastal and non-coastal regions in North America. However, the mechanism of saline adaptation in S. helvola remains unclear. A transcriptome profiling would facilitate dissecting the underlying molecular mechanisms in salinity-adapted S. helvola. In this study, we reported the RNAseq analyses of two genotypes (a salt-tolerant beach genotype and a salt-sensitive inland genotype) of S. helvola stressed with salt. S. helvola plants were grown in pots and treated with half lethal-guided dose of NaCl solution for 3h, 24h, and 7d. The plants supplied with the same amount of water were used as controls. The whole roots sampled from the three time points were equally pooled as one biological replicate, and three replicates were used for library construction and transcriptome sequencing on Illumina Hiseq 2500. The comparative analyses of root transcriptomes presented here provides a valuable resource for discovery of genes and networks involved in salt tolerance in S. helvola.

ORGANISM(S): Strophostyles helvola

PROVIDER: GSE121143 | GEO | 2018/12/06

REPOSITORIES: GEO

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Project description:Background: Soil salinity is a major abiotic stress factor that limit agricultural productivity worldwide, and this problem is expected to grow in the future. Common bean (Phaseolus vulgaris L.) is an important protein source in developing countries is highly susceptible to salt stress. To understand the underlying mechanism of salt stress responses, transcriptomics, metabolomics, and ion content analysis were utilized for response comparison of salt-tolerant and salt-susceptible common bean genotypes in saline conditions. Results: Transcriptome analysis has revealed that the tolerant genotype had increased photosynthesis in saline conditions while the susceptible genotype acted in a contrasting way. The chlorophyll content measurements have backed up this result with increase in tolerant and decrease in susceptible genotype. Transcriptome also displayed a more active carbon and amino acid metabolism for the tolerant genotype as well. Analysis of primary metabolites with GC-MS demonstrated the boosted carbohydrate metabolism in the tolerant genotype with increased sugar content as well as better amino-acid metabolism with the accumulation of glutamate and asparagine and hinted a lowered photorespiration level for the tolerant one. Accumulation of lysine, valine, and isoleucine in the roots of the susceptible genotype suggested a halted stress response pathway. According to ion content comparison, the tolerant genotype managed to block accumulation of Na+ in the leaves while accumulating significantly less Na+ in the roots compared to susceptible genotype. K+ levels increased in the leaves of both genotype and the roots of the susceptible one but dropped in the roots of the tolerant genotype. Additionally, Zn+2 and Mn+2 levels were also dropped in the tolerant roots, while Mo+2 levels were significantly higher in all tissues in both control and saline conditions for tolerant genotype. Conclusion:The results of the presented study have demonstrated the differences in contrasting genotypes and thus provide valuable information on the pivotal molecular mechanisms underlying salt tolerance mainly in common bean, but for all crops.

Project description:Six months old seed grown under well-watered greenhouse conditions were used in the current study. The seedlings of the two wild emmer wheat genotypes were vernalized on a moist germination paper (Hofman Manufacturing, Inc, Jefferson, OR, USA) for three weeks in the dark at 4oC, followed by three days acclimation at 24oC. Seedlings were then transplanted into 5L pots containing a mixture of pure quartz sand and peat (4:1 v/v), supplemented with a slow release fertilizer (2 g/L, Osmocote® Standard 14-14-14, Scotts-Sierra Horticulture, Marysville, OH, USA) and by a weekly application of 100 ml/pot of 0.5X Murashigi and Skoog growth solution (Sigma Chemical Co., St Louis, MO, USA). Pots were placed in a screen-house under natural winter conditions (December to February; 5-18°C) in Haifa, Israel (Mt. Carmel; 35o01′ E, 32o45′ N; 480 m above sea level) for 10 weeks and irrigated daily by a drip irrigation system. Three weeks prior to application of terminal drought stress, the pots were transferred to a controlled environment greenhouse (22/18 oC; 12 h day/12 h night) in order to prevent rainfall during the drought experiment. Five pots per genotype served as biological replicates, plants of three pots of each genotype/treatment were used for transcriptome study whereas two additional pots were used for quantitative PCR analysis (altogether five biological replicated of were used for quantitative real time PCR). Three individual plants were grown in each pot, one plant was used for measurements of leaf relative water content (RWC) (Barrs and Weatherley 1968), whereas the other two plants were used for sampling of flag leaf tissue for RNA extraction. Terminal drought stress (D) was applied at inflorescence emergence stage (Zadoks 50-60), (Zadoks et al. 1974), after emergence of 1-2 spikes in all biological replicates of both genotypes. The time from transplanting to inflorescence emergence stage was not significantly different between genotypes (70 days in the S genotype and 72 days in the R genotype). Drought stress application initiated after irrigation with excess amount of water in order to assure that all pots start the experiment at the same soil water capacity. Drought stress was imposed by withholding water for eight days until stress symptoms (i.e. leaf rolling and wilting symptoms) were visible in plants of both genotypes. Stress symptoms were more visible in the S genotype, however, leaf relative water content (RWC), measured after eight days of drought stress was low but was not significantly different between the two genotypes (49.68%±1.48 in the R genotype and 53.34%±1.83 in the S genotype). The well-watered control (C) plants were irrigated daily by ample amount of water. Flag leaf tissues of drought-stressed plants and well-watered control plants were harvested, immediately frozen in liquid nitrogen, and stored at -80ºC for RNA extraction. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Krugman Tamar. The equivalent experiment is TA33 at PLEXdb.]

Project description:Common bean (Phaseolus vulgaris L.) is the most consumed grain legume in developing countries in Latin America and Sub-Saharan Africa1. Like other legumes, common bean seeds are rich in protein, carbohydrates, fibers and other health-promoting phenolic compounds thus being vital for food security and income source for local small farmers2. Seed quality traits depend on accumulation of various storage molecules during the seed development (SD) process and influenced by the genotype and adaptive changes to environment3. Concerning common bean, there is still a lack of a deeper molecular knowledge of SD that is hampering the development of new biotech approaches for seed trait modulation and could timely address challenges of agriculture or industry. Our present work aims to unravel the molecular mechanisms underlying SD using a proteomic approach. To achieve this goal, we characterized SD in terms biomass, seed length and weight in the genotype SER16, one of the most promissory drought-resistant release of the CIAT-CGIAR. Seed samples were collected at the 4 main SD stages: Late-Embryogenesis (10 days after anthesis, d.a.a.), Early (20 d.a.a.) and Late Maturation (30 d.a.a.) and Desiccation (40 d.a.a.). The analysis of bean proteome was conducted using a gel-free proteomic analysis (LC-MS/MS) under the scope of EU-FP7-PRIME-XS project. A total of 410 unique proteins were differentially expressed throughout the 4 major seed development stages, in which most of the identified proteins belong in the ‘protein metabolism’ (31,98%) functional category, that includes synthesis, regulation, folding. Other functional categories are represented such as carbohydrate and lipid metabolism (11,26%) and stress/defense and redox metabolism (11,04%). We identified 93 proteins were unique to the first (10-20 d.a.a.), 22 to the second (20-30 d.a.a.) and 40 to the last (30-40 d.a.a.) phase transition, reflecting the major biological processes occurring at this specific seed developmental stage. This study will contribute to reveal key metabolic pathways and mechanisms with potential role in modulating common bean seed development and quality traits.

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Transcriptome profiling of a beach-adapted wild legume for dissecting novel mechanisms of salinity tolerance

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