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| 題 名 | 臺灣市售飼料玉米抗嘉磷塞基因特性及檢測利用之探討=Characterization and Detection of the Glyphosate-resistant Gene in Corn Products in Taiwan |
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| 作 者 | 袁秋英; 謝玉貞; 林李昌; 蔣慕琰; | 書刊名 | 作物、環境與生物資訊 |
| 卷 期 | 3:1 民95.03 |
| 頁 次 | 頁20-32 |
| 分類號 | 434.11 |
| 關鍵詞 | 基因改造生物體; 玉米; 嘉磷塞抗藥性; CP4 EPSPS基因; 修飾之玉米抗嘉磷塞EPSPS基因; 複合-聚合酶鏈鎖反應; GMO; Corn; Glyphosate resistance; CP4 EPSPS gene; Modified EPSPS of maize; Multiplex-PCR; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 摘要 於2005 年1 月至2 月期間收集一般雜糧 店販售之飼料玉米,測定其種子發芽及幼株 對嘉磷塞之反應,並由抗性植株葉片中分離 核酸,探討抗藥基因之特性。將市售玉米於 溫室內播種,於4 葉齡幼苗期噴施嘉磷塞 (41% 溶液)1.8 kg ha-1,施藥後5~7 日,多數 玉米植株呈現傷害徵狀,並於2 週內枯死, 14 組測定樣品中均有抗藥性之植株,其所佔 比率介於0.9~3.7%之間,應為抗嘉磷塞之轉 基因玉米。再以嘉磷塞次致死劑量0.5 kg ha-1 處理植株,區別感抗植株後,進一步抽取玉 米幼葉之genomic DNA,由花椰菜嵌紋病毒 啟動子(CaMV 35S promoter)、水稻actin 基 因啟動子及nopaline synthase 基因之終結 子(NOS terminator)核酸序列設計兩組引 子,進行聚合鏈鎖反應,結果具抗藥之疑 似轉基因玉米可分別擴增約2.0 及2.6 kb 片 段或2.1 kb 之單一片段,非抗藥植株則無此 等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,顯示2.0 PCR 產物含水稻actin promoter、阿拉伯芥 ribulose bisphosphate carboxylase/ oxygenase small subunit (RuBisCo SSU)之 chloroplast transit peptide (CTP) 、CP4 EPSPS 及NOS terminator 片段。2.6 kb 之基 因構築為 35S promoter、玉米熱休克蛋白 (SHP70) 基因、CP4 EPSPS 及NOS terminator 片段。2.1 kb 片段則由水稻actin promoter、向日葵RuBisCo SSU 之CTP、玉 米RuBisCo small subunit 之CTP、修飾之 玉米抗嘉磷塞EPSPS 及 NOS terminator 等 片段組成。顯示此2 種玉米為似抗嘉磷塞之 NK603 及GA21 品系。經由專一性引子之設 計及multiplex PCR 分析結果,可分別檢出4 及11 株抗嘉磷塞玉米為似NK603 及GA21 品系。市售之玉米粉、玉米仁、爆米花及玉 米罐頭等食品以multiplex PCR 分析,可檢 出含抗嘉磷塞基因之玉米物料。 |
| 英文摘要 | ABSTRACT We germinated corn seeds collected from retail shops in central Taiwan and tested seedling tolerance against glyphosate. Of all 14 samples tested, seedlings survived herbicide treatment (1.8 kg ha-1 foliar spray) ranged from 0.9-3.7%. These indicated that a small portion of marked feed corn was genetically modified for glyphosate tolerance. We extracted genomic DNA from leaves of seedlings survived low rate of glyphosate spray (0.5 kg ha-1), and conducted PCR with two sets of primers ( CaMV 35S promoter/NOS terminator, rice actin promoter/NOS terminator). PCR amplified fragments were 2.0 and 2.1 kb for promoter using rice actin and 2.6 kb using 35S. Nucleic acid sequences of these fragments were analyzed after conjugation and transformation. Comparison on nucleic and amino acid (NCBI, blast analysis) suggested that components of PCR fragments were: 1) actin promoter, chloroplast transit peptide (CTP) of RuBisCo small subunit from Arabidopsis thaliana, CP4 EPSPS and NOS terminator for 2.0 kb fragment; 2) 35S promoter, heat shock protein from maize (SHP70), CP4 EPSPS and NOS terminator for 2.6 kb fragment; 3) Rice actin promoter, CTP of RuBisCo small subunit from Helianthus annuus, CTP of RuBisCo small subunit from corn, modified EPSPS from corn and NOS terminator for 2.1 kb product. These results also suggested that genetically modified corn lines NK603 and GA21 were in our samples. Multiplex PCR using sequence based primers were conducted on seedlings and corn products. Analysis of 15 glyphosate tolerant seedlings showed that 4 and 11 were NK603 and GA21, respectively. Gene fragments, glyphosateresistant related, were detected in 11 of 17 corn products that sampled in local market. Positive identified products included flour, kernel (fresh, canned), popcorn and corn flake. |
本系統中英文摘要資訊取自各篇刊載內容。