IDENTIFICATION AND MOLECULAR CLONING OF ABIOTIC STRESS TOLERANT GENE(S) AND STRESS INDUCED BIOCHEMICAL CHANGES IN LENTIL (Lens culinaris)

Lentil (Lens culinaris) is an important cool season food legume, encounters numerous biotic and abiotic stresses as it contains asset of genes/proteins which helps this crop to overcome abiotic stresses and identification of stress inducible genes/proteins are an important area of research. Plant productivity is adversely affected by nature’s wrath in the form of various biotic and abiotic stresses. They cause losses worth hundreds of million dollars each year due to reduction in crop productivity and crop failure. In fact, they threaten the sustainability of agricultural industry. In the present study, primers for two abiotic stress tolerant genes namely Inositol and Alcohol dehydrogenase (ADH) were designed by Fast-PCR and validated by Primer-BLAST online tool. Stress treatment at four water potentials (-3.0, -5.0, -7.5 and -10.0 bar) was given followed by RNA isolation. Inositol and ADH were identified through PCR followed by gel elution and cloning in pGEM-T easy and pTZ57R/T vectors respectively. Confirmed clones were sequenced and obtained 1042 bp (Inositol) & 1343 bp (ADH) and analysed through bioinformatics software’s viz. BioEdit, ORF Finder, Mega5, Expasy Protparam tool. Further, biochemical studies in PEG-6000 induced stress showed increased MDA and proline content whereas decreased carbohydrate and protein content as compared to water treated control.