Isolation, molecular characterization and cloning of ect A gene from Halomonaselongata

Abstract

Salinity, an everlasting threat to agricultural production worldwide, affects almost every aspect of the physiologyof plants. The present situation calls for the development of salt tolerant transgenic lines to combat this abiotic stress. Inthis background, the investigation was conducted to isolate the ectA gene responsible for salt tolerance from Halomonaselongata at IABT, Dharwad, to develop transgenic tomato lines by cloning and transferring ectA gene, an ectoine derivativecapable of enzymatic action for the production of acetyl-diaminobutyric acid, gene ectA is involved in maintaining theosmotic balance of plants. The bacterial strains AUDI series was isolated from marine samples along the West-Cost of Indiawhich showed salinity tolerance up to 20 per cent salt concentration. AUDI8 isolate from the PCR positive AUDI seriesstrains was selected for the presence of ectA gene. A 579 bp PCR product containing ectA gene was amplified using ectA genespecific forward and reverse primers. The amplified product was cloned into pTZ57R/T and it was named as pDBJ26 andsequencing of this construct shows 99.66 per cent similarity with other isolates in NCBI (LC168837.1 and AF031489.1).