Studies on Agrobaterium-mediated gene transformation of Oncidium ‘Gower Ramsey’

• Main Authors: Shu-Jung Yang, 楊淑蓉
• Other Authors: Li-Jen Liao
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/23637710894493147633

碩士 === 國立高雄師範大學 === 生物科學研究所 === 91 === Abstract This study transformed Oncidium ’Gower Ramsey’ with Agrobacterium tumefaciens carrying flower color genes - chalcone synthase (CHS) and dihydroflavonol 4- reductase (DFR) and antidisease gene - sweet pepper ferredoxin-like protein (sAP1). Newly formed PLBs of 6-8 weeks cultured were infected by Agrobacterium tumefaciens EHA105. After proliferation for 2 months, the DFR and CHS transformed PLBs and shoots were transferred to MT selection medium containing 30 mg/l hygromycin. After 4 weeks, no PLBs were survival. The Ubi-sAP1 transformed PLBs were selected by Erwinia carotovora subsp. carotovona (ECC) for 2 weeks. Nine out of 118 transformed PLBs survived. The transformation rate is 7.3 % estimated by ECC selection. Moreover, 35S-sAP1, Ubi-CyMV and Ubi-ORSV transformed PLBs which were offered by Lin were transferred into KC medium containing 25 mg /l hygromycin for selection. The putative transgenic PLBs were tested by GUS histochemical staining and only light blue color was shown. As to GFP detection, strongly autofluorescence of plants interfered our observation. The 35S-sAP1 putative transformant PLBs were inoculating with ECC. After 2 weeks, it had 65 % of resistant PLBs. The ‘escape’ phenomenon caused by the lower concentration of hygromycin or the short period of selection, presumably. However, it had been used for untransformed plants to be the control.