Optimisation of genetic transformation procedure for watermelon (Citrullus lanatus Thumb.)

Authors

  • Nguyen Thi Thanh Nga VAST
  • Ho Manh Tuong Institute of Biotechnology, VAST
  • Pham Thi Van Institute of Biotechnology, VAST
  • Nguyen Tuong Van Institute of Biotechnology, VAST
  • Chu Hoang Ha Institute of Biotechnology, VAST
  • Le Tran Binh Institute of Biotechnology, VAST

DOI:

https://doi.org/10.15625/0866-7160/v34n3.2473

Keywords:

Agrobacterium, Citrullus lanatus, gene gus, genetic transformation, kanamycin

Abstract

In this study, the binary vector pCB-gusplus carrying nptII and gus - intron genes as selectable and reportable markers was used to optimize Agrobacterium-mediated gene transformation protocol of four Vietnam watermelon lines. Agrobacterium tumefaciens CV58 habouring pCB-gusplus in the concentration of OD600 0.6-0.8 was infected into 5-7 day cotyledons for 30 minutes and co-cultured for 3 days. The transformed leave materials was cultured in the selectable medium containing MS salts and vitamins supplemented with 1.5 mg/l BAP, 0.5 mg/l IBA, 200 mg/l kanamycin và 500 mg/l cefotaxime. Regenerated shoots was rooted in MS medium added with 0.2 mg/l IBA, 100 mg/l kanamycin and 250 mg/l cefotaxime There was a significant difference in the transformation frequency among watermelon lines. F1 TG939 line showed the transformation frequency of 1.87%), following F9 Tieulong - Thanglong of 1.85%, F1 Kinhkong and F9 Binhthuan lines showed the transformation frequyency of 0%. The analysis of the obtained lines using GUS histochemical assay and PCR reaction with specific primers of ntpII gene had confirmed the presence of gus gene into 5 transgenic lines.

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Published

03-10-2012

How to Cite

Thanh Nga, N. T., Tuong, H. M., Van, P. T., Van, N. T., Ha, C. H., & Binh, L. T. (2012). Optimisation of genetic transformation procedure for watermelon (Citrullus lanatus Thumb.). Academia Journal of Biology, 34(3), 389–396. https://doi.org/10.15625/0866-7160/v34n3.2473

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