Gene transfer in Panax ginseng C.A. Meyer and Drimiopsis kirkii Baker through particle gun bombardment

• Main Authors: Su,Tsung-Jen, 蘇宗振
• Other Authors: Chang,Wei-Chin
• Format: Others
• Language: zh-TW
• Published: 1993
• Online Access: http://ndltd.ncl.edu.tw/handle/55018687301980062928

碩士 === 國立臺灣大學 === 園藝學系 === 81 === In the present studies, embryogenic callus of Panax ginseng C.A. Meyer and leaves of Drimiopsis kirkii Baker were used for gene transformation with a reporter gene (β-glucuronidase gene ,GUS gene) by using particle bombardment techniques. Bombardment explants were then cultured on MS (Murashige & Skoog) medium for embryoid induction. The regenerated embryoids, plantlets and the adjacent callus tissue were then examined to demonstrate the exi- tence of exogenous GUS gene. Under control of the CaMV 35S promotor, the GUS gene con- tained pBI221 (3 kb) were coated to gold particles (diameter 1.6μｍ) and were delivered to targey explants by a setup (PDS1000/ He) od Bio- Rad.Optimal bombardment conditions were found around 650 psi (pounds square inch), 1/4-inch Gap, 1 spacer ring (8 mm) , 25-inch Hg vacuum, 6-cm sample distance and 2 shots. Bombarded explants of P. ginseng C.A.Meyer and D. kirkii Baker expressed the foreign gene activity as checked with X-gluc assays two day the bombardment. Stably expression of GUS gene was found in leaf explants of D. kirkii Baker. However, only temporary expression of GUS gene activity occurred in the bombarded explants of Panax ginseng C.A.Meyer. The pBI121 (13 kb) plasmids were also used for gene trans- formation. Leaves of D. kirkii Baker were transformed with pBI 121 plasmids constructing the GUS and NPT II genes. The bombard- ment coditions were as same as the pBI221 treatment. Transformed explants were cultured on MS medium containing 150 mg/l Kanamy- cin and then produced white embryoids.