Characterization of novel regulators for heat stress tolerance in tomato from Indian sub-continent
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ABSTRACT:
PROVIDER: PRJEB45906 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR6125977_1.fastq.gz | Fastqsanger.gz | |||
| ERR6125977_2.fastq.gz | Fastqsanger.gz |
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Project description:The footprint of tomato cultivation, a cool region crop that exhibits heat stress (HS) sensitivity, is increasing in the tropics/sub-tropics. Knowledge of novel regulatory hot spots from varieties growing in the Indian sub-continent climatic zones could be vital for developing HS-resilient crops. Comparative transcriptome-wide signatures of a tolerant (CLN1621L) and sensitive (CA4) cultivar pair shortlisted from a pool of varieties exhibiting variable thermo-sensitivity using physiological-, survival- and yield-related traits revealed redundant to cultivar-specific HS regulation. The antagonistically expressing genes encode enzymes and proteins that have roles in plant defence and abiotic stresses. Functional characterization of three antagonistic genes by overexpression and silencing established Solyc09g014280 (Acylsugar acyltransferase) and Solyc07g056570 (Notabilis) that are up-regulated in tolerant cultivar, as positive regulators of HS tolerance and Solyc03g020030 (Pin-II proteinase inhibitor), that are down-regulated in CLN1621L, as negative regulator of thermotolerance. Transcriptional assessment of promoters of these genes by SNPs in stress-responsive cis-elements and promoter swapping experiments in opposite cultivar background showed inherent cultivar-specific orchestration of transcription factors in regulating transcription. Moreover, overexpression of three ethylene response transcription factors (ERF.C1/F4/F5) also improved HS tolerance in tomato. This study identifies several novel HS tolerance genes and provides proof of their utility in tomato thermotolerance.
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Project description:Texas tomato production is vulnerable to extreme heat in the spring-summer cropping period, which is exacerbated by the lack of superior genetic materials that can perform well in such environments. There is a dire need for selecting superior varieties that can adapt to warm environments and exhibit high yield stability under heat stress conditions. This research aimed at identifying heat-tolerant varieties under heat-stress conditions in controlled and open-field environments and was carried out in three stages. For the first experiment, 43 varieties were screened based on yield responses in natural open-field environment. From those, 18 varieties were chosen and exposed to control (greenhouse: 26/20 °C) and constant heat-stress (growth-chamber: 34/24 °C) conditions for three months. Measurements were done for chlorophyll fluorescence, chlorophyll content (SPAD), plant height, stem diameter and heat injury index (HII). The last experiment was conducted in an open field with a pool of varieties selected from the first and second experiments. Leaf gas exchange, leaf temperature, chlorophyll fluorescence, SPAD value, electrolyte leakage, heat injury index and yield were assessed. From the combined studies, we concluded that heat-tolerant genotypes selected by using chlorophyll fluorescence and HII in controlled heat-stress conditions also exhibited heat-tolerance in open-field environments. Electrolyte leakage and HII best distinguished tomato varieties in open-field environments as plants with low electrolyte leakage and HII had higher total yield. 'Heat Master,' 'New Girl,' 'HM-1823,' 'Rally,' 'Valley Girl,' 'Celebrity,' and 'Tribeca' were identified as high heat-tolerant varieties. Through trait correlation analysis we provide a better understanding of which traits could be useful for screening and breeding other heat-tolerant tomato varieties.
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Project description:BackgroundHigh temperature is a major environmental stress that limits plant growth and agriculture productivity. Mitogen-activated protein kinases (MAPKs) are highly conserved serine and threonine protein kinases that participate in response to diverse environmental stresses in plants. A total of 16 putative SlMAPK genes are identified in tomato, and SlMAPK3 is one of the most extensively studied SlMAPKs. However, the role of SlMAPK3 in response to heat stress is not clearly understood in tomato plants. In this study, we performed functional analysis of SlMAPK3 for its possible role in response to heat stress.ResultsqRT-PCR analyses revealed that SlMAPK3 relative expression was depressed by heat stress. Here, wild-type (WT) tomato plants and CRISPR/Cas9-mediated slmapk3 mutant lines (L8 and L13) were used to investigate the function of SlMAPK3 in response to heat stress. Compared with WT plants, slmapk3 mutants exhibited less severe wilting and less membrane damage, showed lower reactive oxygen species (ROS) contents, and presented higher both activities and transcript levels of antioxidant enzymes, as well as elevated expressions of genes encoding heat stress transcription factors (HSFs) and heat shock proteins (HSPs).ConclusionsCRISPR/Cas9-mediated slmapk3 mutants exhibited more tolerance to heat stress than WT plants, suggesting that SlMAPK3 was a negative regulator of thermotolerance. Moreover, antioxidant enzymes and HSPs/HSFs genes expression were involved in SlMAPK3-mediated heat stress response in tomato plants.
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