| Summary: | 碩士 === 亞洲大學 === 保健營養生技學系碩士班 === 96 === • This research is to evaluate the pollen flow of transgenic papaya in the open field and to develop a transgenic papaya detection and identification method based on molecular markers. The molecular identification method was carried out by designing a set of primers in the coding area of Papaya ringspot virus (PRSV) coat protein gene and npt II antibiotic gene and then uses polymerase chain reaction (PCR) method to amplify a specific product for transgenic papaya detection and identification. The transgenic papayas used in this trail are hermaphroditic plants (16-0-1) as the pollen donor source while female plants (18-2-4) as a receptor, those seedlings were provided by Dr. Yeh, Shyi-Dong and planted in the isolation field of Taiwan Agricultural Research Institute. We sampled the fruits of female plants located in different directions and distances, and recorded their fruit weights and numbers of seed in each fruit. Thirty seeds in each one fruit were sampled for determine their genotype and segregation rate by PCR method. The spreading range and direction of pollen were estimated by analyzing seeds number and their genotypes. The result shows the spreading range of pollen from hermaphroditic plants (16-0-1) declined with the increase the distance of the female plants (18-2-4) from hermaphroditic, but there was no significant relationship by the statistic analysis. Furthermore, according to our field observation, the insect like honeybee, were considered an important pollinator instead of wind. There was a high frequency of gene flow (38.5%) at 2 m from central transgenic papaya as pollen donor, 5 m and 10 m are 15.3% and 14.8% respectively, the gene flow frequency is steep decline as the distance increased. We discovered that the segregation rate of 16-0-1 RB flanking sequence was low when the distance increased, probably due to the reason that the spread range was low with the pollen contained 16-0-1 RB flanking sequence. According to our results, it was concluded that insect affects the pollen flow of transgenic papaya, and the gene flow of distance can be up to 10 m.
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