Identification and isolation of salt-stress-responsive transcripts from Gossypium arboreum L.

Authors: MUHAMMAD NAVEED SHAHID, ADIL JAMAL, BUSHRA RASHID, BEENISH AFTAB, TAYYAB HUSNAIN

Abstract: Salinity has adverse effects on plant development and ultimately reduces production. Very few reports on the genes induced by salt stress in Gossypium arboreum are available. In this report salt-stress-responsive genes were screened from cotton leaves using the differential display (DD) technique, and gene expression was compared under control and salt stressed conditions. By using a 107 primer combination, 25 gene fragments were found to be up-regulated in response to salt stress. Out of 25 induced fragments, 12 were rejected as false positives after reamplification and quality control assay. The remaining 13 fragments were selected for cloning and genetic transformation. Sequence analysis using GenBank databases revealed that 5 fragments ranging from 300 to 600 bp have significant homology to well-known proteins (i.e. protein kinase, proton gradient regulation, yeast cadmium factor, proteinase inhibitors, and expressed protein). Real time PCR studies confirmed over-expression of the identified transcripts in salt stressed samples as compared to control. These studies will provide insight into the role of these genes in the comprehensive molecular mechanisms of salt tolerance in agriculturally important crops.

Keywords: Cotton (Gossypium arboreum), differential display, salt-stress-responsive transcript

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