A validation study by integrated analysis of physiological, biochemical, and meta-gene expression responses to drought stress in sorghum (Sorghum bicolor L.).

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Sorghum (Sorghum bicolor L.) is a prominent cereal known for its high photosynthetic efficiency and biomass production, serving as a source of food, animal feed, fiber, and biofuels. This study aims to validate identified meta-genes associated with drought stress in sorghum. Two cultivars, Mansour (drought-tolerant) and Pegah (drought-susceptible), were subjected to drought stress at four levels (25%, 50%, 75%, and 100% of field capacity [FC]( During the 4-5 leaf stage in a greenhouse in 2021. The physiological and molecular responses of the sorghum samples were evaluated at 24, 48, 72, and 96h post-treatment. The expression of five meta-genes was analyzed to validate these candidate genes related to drought stress tolerance in sorghum. Analysis of variance indicated that the main effects of drought, cultivar, and sampling time, as well as their interactions, had highly significant effects (P < 0.01) on most physiological and biochemical traits. The relative expression of the genes SORBI_3002G225100, SORBI_3003G332200, SORBI_3003G368300, SORBI_3010G081800, and SORBI_3004G293500 increased over time under drought stress. Proline levels, ion leakage, soluble sugars, and the activities of catalase, peroxidase, ascorbate peroxidase, and superoxide dismutase enzymes increased with the intensity of drought stress and over time. Conversely, the levels of chlorophyll a and b, carotenoids, RWC, leaf surface area, and protein content decreased under drought conditions. These results confirm the relevance of these genes in conferring drought stress tolerance in sorghum. This research provides new finding into the physiological processes and biochemical activities, alongside the validation of meta-gene expression involved in drought stress, further advancing our understanding of molecular mechanisms of the reaction of sorghum to drought stress.

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  • Addendum
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  • 10.1371/journal.pone.0197017
Correction: Cross-species multiple environmental stress responses: An integrated approach to identify candidate genes for multiple stress tolerance in sorghum (Sorghum bicolor (L.) Moench) and related model species.
  • May 3, 2018
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[This corrects the article DOI: 10.1371/journal.pone.0192678.].

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Cross-species multiple environmental stress responses: An integrated approach to identify candidate genes for multiple stress tolerance in sorghum (Sorghum bicolor (L.) Moench) and related model species
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BackgroundCrop response to the changing climate and unpredictable effects of global warming with adverse conditions such as drought stress has brought concerns about food security to the fore; crop yield loss is a major cause of concern in this regard. Identification of genes with multiple responses across environmental stresses is the genetic foundation that leads to crop adaptation to environmental perturbations.MethodsIn this paper, we introduce an integrated approach to assess candidate genes for multiple stress responses across-species. The approach combines ontology based semantic data integration with expression profiling, comparative genomics, phylogenomics, functional gene enrichment and gene enrichment network analysis to identify genes associated with plant stress phenotypes. Five different ontologies, viz., Gene Ontology (GO), Trait Ontology (TO), Plant Ontology (PO), Growth Ontology (GRO) and Environment Ontology (EO) were used to semantically integrate drought related information.ResultsTarget genes linked to Quantitative Trait Loci (QTLs) controlling yield and stress tolerance in sorghum (Sorghum bicolor (L.) Moench) and closely related species were identified. Based on the enriched GO terms of the biological processes, 1116 sorghum genes with potential responses to 5 different stresses, such as drought (18%), salt (32%), cold (20%), heat (8%) and oxidative stress (25%) were identified to be over-expressed. Out of 169 sorghum drought responsive QTLs associated genes that were identified based on expression datasets, 56% were shown to have multiple stress responses. On the other hand, out of 168 additional genes that have been evaluated for orthologous pairs, 90% were conserved across species for drought tolerance. Over 50% of identified maize and rice genes were responsive to drought and salt stresses and were co-located within multifunctional QTLs. Among the total identified multi-stress responsive genes, 272 targets were shown to be co-localized within QTLs associated with different traits that are responsive to multiple stresses. Ontology mapping was used to validate the identified genes, while reconstruction of the phylogenetic tree was instrumental to infer the evolutionary relationship of the sorghum orthologs. The results also show specific genes responsible for various interrelated components of drought response mechanism such as drought tolerance, drought avoidance and drought escape.ConclusionsWe submit that this approach is novel and to our knowledge, has not been used previously in any other research; it enables us to perform cross-species queries for genes that are likely to be associated with multiple stress tolerance, as a means to identify novel targets for engineering stress resistance in sorghum and possibly, in other crop species.

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Post-flowering drought tolerance (stay-green) in grain sorghum (Sorghum bicolor (L.) Moench) is an important agronomic trait in many arid and semiarid environments throughout the world. Stay-green has been associated with increased grain yields, as well as resistance to lodging and charcoal rot disease. Nonetheless, the relative effects of genotype, environment, and genotype × environment interactions are not well understood for this trait; similarly, the relationship between various leaf sugars and stay-green has not been sufficiently evaluated in diverse germplasm. Thus, the goals of this study were to determine the genotype, environment, and genotype by environment (GxE) effects for leaf dhurrin, sugars, and stay-green in ten diverse grain sorghum breeding lines, to evaluate the Pearson’s correlation coefficients (r) between these traits, and to determine entry-mean repeatability (R) for each of these traits. Of the compositional traits studied, we determined that leaf dhurrin had the highest correlation with the stay-green phenotypes (r = −0.62). We found that stay-green sorghum lines contained approximately 2–3 times as much dhurrin as non-stay-green lines, with B1778 containing the highest concentration of dhurrin (84.8 µg/cm2) and Tx7000 containing the least (20.9 µg/cm2). The differences between the environments for several of the traits were high, and all the traits examined had high repeatability (R = 0.89–0.92). These data demonstrate a relationship between leaf dhurrin and the stay-green phenotypes in sorghum, and further study will allow researchers to determine the causal effect that dhurrin has on post-flowering drought tolerance in sorghum.

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  • Research Article
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BackgroundDrought is the most disastrous abiotic stress that severely affects agricultural productivity worldwide. Understanding the biological basis of drought-regulated traits, requires identification and an in-depth characterization of genetic determinants using model organisms and high-throughput technologies. However, studies on drought tolerance have generally been limited to traditional candidate gene approach that targets only a single gene in a pathway that is related to a trait. In this study, we used sorghum, one of the model crops that is well adapted to arid regions, to mine genes and define determinants for drought tolerance using drought expression libraries and RNA-seq data.ResultsWe provide an integrated and comparative in silico candidate gene identification, characterization and annotation approach, with an emphasis on genes playing a prominent role in conferring drought tolerance in sorghum. A total of 470 non-redundant functionally annotated drought responsive genes (DRGs) were identified using experimental data from drought responses by employing pairwise sequence similarity searches, pathway and interpro-domain analysis, expression profiling and orthology relation. Comparison of the genomic locations between these genes and sorghum quantitative trait loci (QTLs) showed that 40% of these genes were co-localized with QTLs known for drought tolerance. The genome reannotation conducted using the Program to Assemble Spliced Alignment (PASA), resulted in 9.6% of existing single gene models being updated. In addition, 210 putative novel genes were identified using AUGUSTUS and PASA based analysis on expression dataset. Among these, 50% were single exonic, 69.5% represented drought responsive and 5.7% were complete gene structure models. Analysis of biochemical metabolism revealed 14 metabolic pathways that are related to drought tolerance and also had a strong biological network, among categories of genes involved. Identification of these pathways, signifies the interplay of biochemical reactions that make up the metabolic network, constituting fundamental interface for sorghum defence mechanism against drought stress.ConclusionsThis study suggests untapped natural variability in sorghum that could be used for developing drought tolerance. The data presented here, may be regarded as an initial reference point in functional and comparative genomics in the Gramineae family.

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Drought stress is a major constraint to sorghum production in Kenya, especially during flowering stage. This study aimed at developing drought tolerant sorghum varieties by transferring the stay green trait that confers drought tolerance in sorghum from a mapped and characterized donor source into an adapted farmer preferred variety. The drought tolerance donor source, E36-1 originally from Ethiopia was backcrossed into a Kenyan farmer-preferred variety, Ochuti until BC2F1 generation and the stay-green Quantitative Trait Loci (QTL) were transferred through Marker Assisted Breeding (MAB) strategy. Five polymorphic Simple Sequence Repeat (SSR) markers were used to select the 3 stay green QTL of E36-1 found in SBI-01, SBI- 07 and SBI-10 linkage groups. In the F1 generation, two of these QTL, were transferred into three genotypes. In the BC1F1 generation, 32 genotypes had at least one QTL incorporated. From a population of 157 BC2F1 progenies, 45 genotypes had incorporated either one or two of the stay-green QTL. Despite a few number of genotypes obtained through the backcrosses, the results showed that stay-green QTL and consequently drought tolerance can be transferred successfully into farmer preferred sorghum varieties through MAB.

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The Promising B-Type Response Regulator hst1 Gene Provides Multiple High Temperature and Drought Stress Tolerance in Rice.
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High temperatures, drought, and salt stresses severely inhibit plant growth and production due to the effects of climate change. The Arabidopsis ARR1, ARR10, and ARR12 genes were identified as negative salt and drought stress regulators. However, in rice, the tolerance capacity of the hst1 gene, which is orthologous to the ARR1, ARR10, and ARR12 genes, to drought and multiple high temperature and drought stresses remains unknown. At the seedling and reproductive stages, we investigated the drought (DS) high temperature (HT) and multiple high temperature and drought stress (HT+DS) tolerance capacity of the YNU31-2-4 (YNU) genotype, which carries the hst1 gene, and its nearest genomic relative Sister Line (SL), which has a 99% identical genome without the hst1 gene. At the seedling stage, YNU demonstrated greater growth, photosynthesis, antioxidant enzyme activity, and decreased ROS accumulation under multiple HT+DS conditions. The YNU genotype also demonstrated improved yield potential and grain quality due to higher antioxidant enzyme activity and lower ROS generation throughout the reproductive stage under multiple HT+DS settings. Furthermore, for the first time, we discovered that the B-type response regulator hst1 gene controls ROS generation and antioxidant enzyme activities by regulating upstream and downstream genes to overcome yield reduction under multiple high temperatures and drought stress. This insight will help us to better understand the mechanisms of high temperature and drought stress tolerance in rice, as well as the evolution of tolerant crops that can survive increased salinity to provide food security during climate change.

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