Somatic Embryogenesis and Gene Transformation with ORSV Coat Protein Gene of Oncidium

• Main Authors: Hui-Ju Shen, 沈惠茹
• Other Authors: Wei-Chih Chang
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/96964097604929504807

碩士 === 國立臺灣大學 === 園藝學研究所 === 93 === Explants of Oncidium Louise Elmore ‘Elsa’ young leaves produced somatic embryos from adaxial surfaces directly without an intervening callus within 3 months when cultured on a 1/2 MS basal medium supplemented 0.3, 3 mg/l TDZ or 3 mg/l BA or 0.3, 1, 3 mg/l zeatin. The better response was found at 3 mg/l TDZ. In this treatment, 55.0％ of leaf explants formed 3.53 somatic embryos. Somatic embryos developed into plantlets (4 cm in height) with several roots and shoots when cultured on a medium devoid of plant growth regulator after 4 months of culture. 1-cm-long leaf tip segments of Oncidium ‘Gower Ramsey’ were cultured on embryo induction medium (containing 0.5 mg/l TDZ). Until the 8th week of induction, excised leaves were bombarded every week by gene gun with plasmid pKyLx7. At the 10th week, embryos were transplanted to a basal medium supplemented with 0.5 mg/l NAA for 4 to 6 weeks. With light supplement 3 to 4 months, embryos could develop to 1-cm-high plants. Selection for putative transgenic plants were accomplished by using 500 mg/l kanamycin 2 weeks. There were 230 putative transgenic plants survived. Survival transgenic plants were randomly selected 21 plants to conform by PCR. However, the result showed there didn’t exist ORSV coat protein gene in 21 putative transgenic plants.