台灣市售抗固殺草基改大豆之篩選及其基因特性分析

Genomic Characterization of Imported Soybean Resistant to Glufosinate in Taiwan

袁秋英(C. I. Yuan) ； 林李昌(L. C. Lin) ； 蔣慕琰(M. Y. Chiang)

《中華民國雜草學會會刊》 32卷1期 (2011/06) Pp. 1-9

https://doi.org/10.6274/WSSROC-2011-032(1)-001

摘要

台灣每年進口大豆約有250萬噸，一般市售的大豆種子尚具有生物活性，本研究於中部零售店收集大豆種子，測定其種子發芽率及幼株對固殺草（glufosinate）除草劑之反應，並由抗性植株葉片中萃取核酸，探討抗固殺草轉基因之構築與特性。將大豆種子於溫室內播種，株齡為3－5葉之幼苗噴施固殺草0.675 kg ai ha^(-1)，施藥後3－4日，多數大豆植株呈現傷害徵狀，並於1週內枯死，12組測定樣品中均有抗藥性之植株，其所佔比率介於0－13%之間，初步推測為抗固殺草之轉基因大豆。進一步抽取大豆幼葉基因組DNA，以CaMV 35S啟動子（promoter）及35S終結子（terminator）核酸序列設計引子，進行PCR反應，結果具抗藥之疑似轉基因大豆可增幅約1.0 kb DNA片段，非抗性大豆則未增幅出此DNA片段。PCR產物經接合、轉型反應及定序，比對核酸及胺基酸序列，結果此PCR產物由35S啟動子（265 bp）、pat 基因（501 bp） 及35S終結子（110 bp）等片段組成，顯示此抗固殺草大豆轉基因組成與LibertyLink （A2704-12）大豆品系者相同，其中抗固殺草的pat基因序列與基改玉米T25及Bt11品系者完全相同。

關鍵字

基因改造生物體 ； 大豆 ； 固殺草抗藥性 ； pat基因

並列摘要

Two point five million tons of soybean seeds are imported annually into Taiwan. Most seeds sampled from local shops were found to be viable. Soybean seedlings were established in greenhouse and applied 0.675 kg ai ha^(-1) of glufosinate to the leaves of these plants. Sensitive seedlings showed symptoms of injury in 3-4 days and died within one week. Glufosinate-resistant seedlings accounted for 6-13% of total samples. The plants survived from glufosinate treatment were subjected to PCR detection and genomic characterization. PCR assay of genomic DNA extracted from soybean leaves, using the CaMV 35S promoter and 35S terminator as primers, produced a 1.0 kb fragments. DNA sequencing using PCR products showed that the fragments included 35S promoter (265 bp), pat (501 bp) and 35S terminator (110 bp) were same as of the transgenes of LibertyLink soybeans (A2704-12) from Bayer CropScience.

並列關鍵字

Genetically modified organism (GMO) ； soybean ； glufosinate resistance ； pat gene