Characterization of high-temperature stress-tolerant tomato (Solanum lycopersicum L.) genotypes by biochemical analysis and expression profiling of heat-responsive genes

The survival, biomass, and grain yield of most of the crops are negatively influenced by several environmental stresses. The present study was carried out by using transcript expression profiling for functionally clarifying the role of genes belonging to a small heat shock protein (sHSP) family in pearl millet under high-temperature stress. Transcript expression profiling of two high-temperature-responsive marker genes, Pgcp70 and PgHSF, along with physio-biochemical traits was considered to screen out the best contrasting genotypes among the eight different pearl millet inbred lines in the seedling stage. Transcript expression pattern suggested the existence of differential response among different genotypes upon heat stress in the form of accumulation of heat shock-responsive gene transcripts. Genotypes, such as WGI 126, TT-1, TT-6, and MS 841B, responded positively toward high-temperature stress for the transcript accumulation of both Pgcp70 and PgHSF and also indicated a better growth under heat stress. PPMI-69 showed the least responsiveness to transcript induction; moreover, it supports the membrane stability index (MSI) data for scoring thermotolerance, thereby suggesting the efficacy of transcript expression profiling as a molecular-based screening technique for the identification of thermotolerant genes and genotypes at particular crop growth stages. The contrasting genotypes, such as PPMI-69 (thermosusceptible) and WGI-126 and TT-1 (thermotolerant), are further utilized for the characterization of thermotolerance behavior of sHSP by cloning a PgHSP16.97 from the thermotolerant cv. WGI-126. In addition, the investigation was extended for the identification and characterization of 28 different HSP20 genes through a genome-wide search in the pearl millet genome and an understanding of their expression pattern using the RNA-sequencing (RNA-Seq) data set. The outcome of the present study indicated that transcript profiling can be a very useful technique for high-throughput screening of heat-tolerant genotypes in the seedling stage. Also, the identified PgHSP20s genes can provide further insights into the molecular regulation of pearl millet stress tolerance, thereby bridging them together to fight against the unpredicted nature of abiotic stress.

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Differential modulation of heat inducible genes across diverse genotypes and molecular cloning of a sHSP from Pearl millet [Pennisetum glaucum (L.) R. Br.]

10.1101/2020.02.26.966184 ◽

2020 ◽

Author(s):

S MukeshSankar ◽

C. Tara Satyavathi ◽

Sharmistha Barthakur ◽

S.P Singh ◽

Roshan Kumar ◽

...

Keyword(s):

Heat Stress ◽

High Temperature ◽

Pearl Millet ◽

Temperature Stress ◽

Expression Profiling ◽

Stability Index ◽

High Temperature Stress ◽

Seedling Stage ◽

Transcript Expression ◽

Membrane Stability Index

AbstractEnvironmental stresses negatively influence survival, biomass and grain yield of most crops. Towards functionally clarifying the role of heat responsive genes in Pearl millet under high temperature stress, the present study were carried out using semi quantitative RT- PCR for transcript expression profiling of hsf and hsps in 8 different inbred lines at seedling stage, which was earlier identified as thermo tolerant/susceptible lines through initial screening for thermo tolerance using membrane stability index among 38 elite genotypes. Transcript expression pattern suggested existence of differential response among different genotypes in response to heat stress in the form of accumulation of heat shock responsive gene transcripts. Genotypes WGI 126, TT-1 and MS 841B responded positively towards high temperature stress for transcript accumulation for both Pgcp 70 and Pghsf and also had better growth under heat stress, whereas PPMI 69 showed the least responsiveness to transcript induction supporting the membrane stability index data for scoring thermotolerance, suggesting the efficacy of transcript expression profiling as a molecular based screening technique for identification of thermotolerant genes and genotypes at particular crop growth stages. As to demonstrate this, a full length cDNA of Pghsp 16.97 was cloned from the thermotolerant cultivar, WGI 126 and characterized for thermotolerance. The results of demonstration set forth the transcript profiling for heat tolerant genes can be a very useful technique for high throughput screening of tolerant genotypes at molecular level from large cultivar collections at seedling stage.

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