模擬GM大油菜與小油菜之雜交後代的鑑別

Identifying the F_1 hybrids of the Simulated GM Brassica napus and Brassica rapa

洪鴻(Hong Hong) ； 林子凱(Tzu-Kai Lin) ； 杜元凱(Yuan-Kai Yu) ； 郭寶錚(Bo-Jein Kuo)

《作物、環境與生物資訊》 13卷2期 (2016/06) Pp. 53-66

https://doi.org/10.30061/CEB.201606_13(2).0001

摘要

油菜的基因改造(genetically modified;GM)技術在1990 年初期開始應用於大油菜(Brassica napus)，有許多重要的農藝性狀相關基因，已經藉由基因改造技術轉入大油菜品系之中。GM 作物所造成的基因流動除對田區雜草管理上產生問題外，亦可能影響生態環境或作物的產銷。有許多因素會對基因流動產生不同的影響，因此對於預測或是預防基因流動有其困難性。本篇利用自大陸地區引進的商業上推廣種植B. napus 品種模擬GM 大油菜，以及臺灣地區常見的小油菜(B.rapa)品種，在溫室種植並進行B. napus 及B.rapa 間人工雜交，取得雜交後代種子。並利用SRAP (sequence-related amplified polymorphism) 之技術， 配合流式細胞儀(Flow CytoMetry; FCM) 分析細胞核酸含量，輔以外表性狀之觀察，用來鑑別雜交後代種子。結果發現，後代外表型有傾向大油菜的現象，經流式細胞儀分析雜交後代細胞內核酸含量，發現含量位於兩親本之間；SRAP 分析的部分，從30 組引子組合當中選擇了7 組引子組合作為分析之用，在雜交後代中至少有5 個可供鑑別的條帶。最後利用試驗結果對田間自然授粉之種子進行分析，發現無論是基因型或外表型的分析結果，均與人工雜交種子的結果一致。這些結果可應用於監測田間所種植的小油菜是否有被GM大油菜花粉所汙染。

關鍵字

小油菜 ； 大油菜 ； 相關序列增幅多型性 ； 流式細胞儀

並列摘要

The technology of genetically modified (GM) of Brassica oilseed started from Brassica napus in the early 1990s. There are a number of genes related to the important agronomic traits which have been transferred into rapeseed lines by GM technology. Gene flow results in not only weed management problems, but also influences the ecological environment or crops production in the field. The gene flow could be affected by many factors, so the prediction or prevention of gene flow was very difficult. In this study, we employed a B. napus species, which is a commercial variety used to simulate the GM oilseed rape in Mainland China, and a B. rapa species, which is green manure species in Taiwan. All materials were planted in a greenhouse and then implemented the crossing experiments to produce hybrid seeds. In order to confirm the observation of the difference between parents and their offspring, the sequence-related amplified polymorphism (SRAP) molecular maker and the flow cytometric (FCM) were used to analyze them. As results indicated, phenotypes of hybrids were more close to B. napus and the DNA content of hybrids was between that of their parents. In the SRAP study, seven primer pairs from thirty SRAP primer pairs to analyze the hybrids. It was found that there were at least five polymorphic molecular markers could be used to identify hybrids. Finally, a real field-level experiment was employed to confirm that the results of hybrid seeds from artificial and open-pollination are the same. These results could be used to monitor whether B. rapa in the field has been contaminated by the GM B. napus.

並列關鍵字

Brassica rapa ； Brassica napus ； Sequence-related amplified polymorphism (SRAP) ； Flow CytoMetry (FCM)