Genetic transformation in Curcuma alismatifolia Gagnep., Petunia axillaris and Nicotiana tabacum via Agrobacterium tumefaciens

Mondhon Sanguansermsri, Popkao Buddharak, Somboon Anuntalabhochai, Nopmanee Topoonyanont, Peerasak Chaiprasart


Gus gene was transformed into Siam tulip (Curcuma alismatifolia Gagnep.),petunia (Petunia axillaries) and tobacco (Nicotiana tabacum) using 2 types of plasmid(pSCV1.6 and pBI121) in Agrobacterium tumefaciens. Two days after co-cultivation withbacteria, C. alismatifolia Gagnep. were grown on a selective medium with 50 mg/lkanamycin (RM1). The GUS gene were detected by GUS histochemical assay and PCRanalysis. The tobacco shoots and petunia shoots were grown on another selective mediumwith 50 mg/l kanamycin (RM2) and then their GUS genes were detected by GUShistochemical assay. In case of C. alismatifolia Gagnep., the blue spots indicating thepresence of GUS gene were found at 0.83%. The gene transformation was also confirmed bythe presence of DNA strip of CaMV 35S promoter primer using PCR analysis. The explantsof P. axillaris and N. tabacum which transformed with pSCV1.6 showed GUS positive at23.33% and 14.4%, respectively. The explants of P. axillaris and N. tabacum whichtransformed with pBI121 showed GUS positive present 13.33% and 3%, respectively. Fortobacco and petunia, although blue spots could be found on all leaf surfaces transformed withplasmid pSCV1.6 and pBI121 but blue spots which found on plasmid pSCV1.6 gave a betterindicate for detecting GUS gene.

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