Construction of transformation vectors and production of transgenic melons expressing the coat protein genes of Zucchini yellow mosaic virus and Papaya ringspot virus type W

• Main Authors: Ming-Der Lin, 林明德
• Other Authors: Hui-Liang Wang
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/21949284726179255176

碩士 === 國立高雄師範大學 === 生物科學研究所 === 94 === Abstract Melon (Cucumis melo L.) is an important crop of Cucurbitaceae. It is susceptible to both Zucchini yellow mosaic virus (ZYMV) and Papaya ringspot virus type watermelon (PRSV-W) which are limiting factors of the production of melon. The application of biotechnological tools in melon breeding have attracted increasing interest and provided a valuable complementary conventional improvement of this species. In constructs of pCAMBIA 1304 or pCAMBIA 2301 binary vector containing ZYMV, PRSV-W or both coat protein gene constructs were mobilized from Escherichia coli into the disarmed Agrobactrium tumefaciens strain EHA 105 by eletroporation. In GUS test, cotyledon via Agrobactrium-mediated gene transformation showed strong GUS transient expression as visualized 4 day after co-cultivation. In melon transformation experiment result, explant regeneration only appeared in kanamycin selection medium but not in hygromycin selection medium. The putative transforants were confirmed by PCR using specific primers to CP gene, 35S promoter and NOS terminator. PCR positive plants of two lines that containing ZMMV-7 CP gene and six lines that containing both CP gene were selected. RT-PCR and ELISA analysis for those transgenic melons showed negative reaction. Virus resistance of the transgenic melons were testing.