A Novel Biochemical Method for Screening of Cold Tolerance in Rice

  • S. K. Agrawal Research Luboraoly for Agricultural Biotechnology and Biochemistry (RLABB) GPO Box 2128, Kathmandu
  • G. K. Agrawal Research Luboraoly for Agricultural Biotechnology and Biochemistry (RLABB) GPO Box 2128, Kathmandu
  • V. P. Agrawal Royal Nepal Academy of Science and Technology (RONAST) GPO Box 3323, Khumaltar

Abstract

The present biochemical method involves quantification of Ribulose- l,5-biphosphate carboxylase/oxygenase (RuBisCo). It is a simple, sensitive, rapid and a non-destructive method for screening of cold tolerance in rice (Oryza sativa L). Using this method, rice lines can be categorized into cold tolerant (CT), moderately cold tolerant (MCT), and cold sensitive (CS). In this method, 12-day old rice seedlings grown at 26°C are treated at 12°C for five days under a 12/12 h light/dark regime, and the level of total RuBisCo is measured by sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE) of leaf protein followed by coomassie staining with subsequent determination of absorbance at 595 nm of the dye eluted from the bands of lage and small subunits (LSU and SSU) of RuBisCo. Verification of this method has been done with 60 rice cultivars of different genotypes including CT, MCT, and CS cultivars.This RuBisCo method provides a convenient, inexpensive, and reliable biochemical procedure for distinguishing rice varieties on basis of cold tolerance andlor sensitivity. This is also the first and only known method till date for screening of cold tolerance in rice