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

Nguyen Thi Thanh Nga, Ho Manh Tuong, Pham Thi Van, Nguyen Tuong Van, Chu Hoang Ha, Le Tran Binh

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.


Keywords


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



DOI: https://doi.org/10.15625/0866-7160/v34n3.2473 Display counter: Abstract : 118 views. PDF (Tiếng Việt) : 152 views.

 

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