STUDY IN VITRO REGENERATION AND TRANSFORMATION SYSTEMS OF CUCUMBER VARIETY CHOKA F1 USING GUS REPORTER GENE

Abstract

This study was performed to establish and develop efficient procedures for Agrobacterium-mediated transformation and regeneration of an inbred cucumber cultivar, which provides a potential protocol to transfer gene of interest into cucumber as well as further research on genome editing in cucumber in Vietnam. In this procedure, cucumber seed sterilization was well performed using 15% NaClO for 15 min. The frequency of multiple shoot induction was 61.76% and the number of regenerated shoots per sample was 2.45 on MS basal medium added 1.5 mg/L BAP and 1 mg/L ABA. The rooting frequency ranged from 45% to 85% and the whole regenerated plants were formed on MS medium supplemented with 20 g/L sucrose. The in vitro regenerated cucumber plants were successfully transferred and grown on vermiculite under greenhouse conditions with a survival rate of 66.67%. These medium components and the regeneration procedure were successfully utilized to transfer gus reporter gene into the Choka F1 variety using the Agrobacterium-mediated method. The GUS staining and PCR analysis using specific primers demonstrated the presence and expression of gus gene in transgenic cucumber lines. The transformation frequency was preliminary recorded at 9.44%.