Proteome dynamics and transcriptome profiling in sorghum [ Sorghum bicolor (L.) Moench] under salt stress
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ORIGINAL ARTICLE
Proteome dynamics and transcriptome profiling in sorghum [Sorghum bicolor (L.) Moench] under salt stress Himani Punia1 · Jayanti Tokas1 · Surina Bhadu1 · Ashok K. Mohanty2 · Preeti Rawat2 · Anurag Malik3 · Satpal4 Received: 19 June 2020 / Accepted: 11 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract Sorghum is a C4 cereal grain crop which is well adapted to harsh environment. It is a potential model for gaining better understanding of the molecular mechanism due to its wider adaptability to abiotic stresses. In this study, protein extraction was standardized using different methods to study the electrophoretic pattern of sorghum leaves under different salinity levels. The extraction of soluble protein with lysis buffer, followed by its clean-up was found to be the most effective method. The different profiles of salt-responsive proteins were analyzed in G-46 and CSV 44F sorghum genotypes based on their tolerance behavior towards salinity. The kafirin level also changed depending upon the concentration and exposure time to salts suggesting the stored proteins as energy source under stress conditions. The relative expression of salt-responsive genes was studied using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) which might be used as a molecular screening tool for identification of salt-tolerant genotypes in affected areas. The validated responses were examined in terms of metabolic changes and the expression of stress-induced proteins—viz. heat shock proteins (hsp) via immunoblotting assay. The results showed that the two sorghum genotypes adopted distinct approaches in response to salinity, with G-46 performing better in terms of leaf function. Also, we have standardized different protein extraction methods followed by their clean-up for electrophoretic profiling. Keywords Electrophoretic profile · Heat shock protein · Polyacrylamide gel electrophoresis · Salinity · Sorghum Abbreviations Act Actin β-ME β-Mercaptoethanol BADH1 Betaine aldehyde dehydrogenase CBB Coomassie Brilliant Blue DTT Dithiothreitol H+-PPase Vacuolar hydrogen pyrophosphatase
HSP Heat shock protein PAGE Polyacrylamide gel electrophoresis P5CS1 Pyrroline-5 carboxylate synthetase 1 PMSF Phenylmethylsulfonylfluoride RT-PCR Reverse transcriptase-polymerase chain reaction
* Himani Punia [email protected]
1
Jayanti Tokas [email protected]
Department of Biochemistry, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar, Haryana 125 004, India
2
Surina Bhadu [email protected]
Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India
3
Department of Seed Science & Technology, College of Agriculture, CCS Haryana Agricultural University, Hisar, Haryana 125 004, India
4
Forage Section, Department of Genetics and Plant Breeding, College of Agriculture, CCS Haryana Agricultural University, Hisar, Haryana 125 004, India
Ashok K. Mohanty [email protected] Preeti Rawat rawatpreeti
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