Characterizing the putative mitogen-activated protein kinase (MAPK) and their protective role in oxidative stress tolerance and carbon assimilation in wheat under terminal heat stress

Ranjeet R Kumar; Kavita Dubey; Kirti Arora; Monika Dalal; Gyanendra K Rai; Dwijesh Mishra; Krishna K Chaturvedi; Anil Rai; Soora Naresh Kumar; Bhupinder Singh; Viswanathan Chinnusamy; Shelly Praveen

doi:10.1016/j.btre.2021.e00597

Characterizing the putative mitogen-activated protein kinase (MAPK) and their protective role in oxidative stress tolerance and carbon assimilation in wheat under terminal heat stress

Biotechnol Rep (Amst). 2021 Feb 5:29:e00597. doi: 10.1016/j.btre.2021.e00597. eCollection 2021 Mar.

Authors

Affiliations

¹ Division of Biochemistry, Indian Agricultural Research Institute, New Delhi, 110012, India.
² ICAR-National Institute for Plant Biotechnology, Pusa Campus, New Delhi, 110012, India.
³ Sher-e-Kashmir University of Agricultural Sciences and Technology, Jammu, 180009, India.
⁴ CABin, Indian Agricultural Statistics Research Institute, New Delhi, 110012, India.
⁵ Centre for Environment Science and Climate Resilient Agriculture (CESCRA), Indian Agricultural Research Institute, New Delhi, 110012, India.
⁶ Division of Plant Physiology, Indian Agricultural Research Institute, New Delhi, 110012, India.

Abstract

Wheat, being sensitive to terminal heat, causes drastic reduction in grain quality and yield. MAPK cascade regulates the network of defense mechanism operated inside plant system. Here, we have identified 21 novel MAPKs through gel-based proteomics and RNA-seq data analysis. Based on digital gene expression, two transcripts (transcript_2834 and transcript_8242) showing homology with MAPK were cloned and characterized from wheat (acc. nos. MK854806 and KT835664). Transcript_2834 was cloned in pET28a vector and recombinant MAPK protein of ∼40.3 kDa was isolated and characterized to have very high in-vitro kinase activity under HS. Native MAPK showed positive correlation with the expression of TFs, HSPs, genes linked with antioxidant enzyme (SOD, CAT, GPX), photosynthesis and starch biosynthesis pathways in wheat under HS. Wheat cv. HD3086 (thermotolerant) having higher expression and activity of MAPK under HS showed significant increase in accumulation of proline, H₂O₂, starch, and granule integrity, compared with BT-Schomburgk (thermosusceptible).

Keywords: 2-DE, 2-dimensional electrophoresis; CAT, Catalase; CD, Conserved domain; DGE, Digital gene expression; HS, Heat stress; HSR, Heat shock response; Heat-Shock response; MAPK, Mitogen-activated protein kinase; Mitogen-Activated protein kinases; Recombinant fusion proteins; SAGs, Stress-associated genes; SAPs, Stress-associated proteins; Starch; Thermotolerance; Triticum.