[1]张秀春　 彭 　 明　 吴坤鑫　 郭丽琼,林俊扬　 林俊芳.利用双 T - DNA 载体系统培育无选择标记转基因大豆*[J].大豆科学,2006,25(04):369-372.[doi:10.11861/j.issn.1000-9841.2006.04.0369]
　Zhang Xiuchun Peng Ming Wu Kunxin Guo Liqiong Lin Junyang Lin Junfang.GENERATING MARKER-FREE TRANSGENIC SOYBEAN PLANTS BY AGROBACTERIUM-MEDIATED TRANSFORMATION WITH DOUBLE T- DNA BINARY VECTOR[J].Soybean Science,2006,25(04):369-372.[doi:10.11861/j.issn.1000-9841.2006.04.0369]
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利用双 T - DNA 载体系统培育无选择标记转基因大豆*

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第25卷 期数: 2006年04期 页码: 369-372 栏目: 出版日期: 2006-11-25
Title:
GENERATING MARKER-FREE TRANSGENIC SOYBEAN PLANTS BY AGROBACTERIUM-MEDIATED TRANSFORMATION WITH DOUBLE T- DNA BINARY VECTOR
作者:
张秀春1　 彭 　 明1　 吴坤鑫1　 郭丽琼1;  2　 林俊扬3　 林俊芳1;  2
(1. 中国热带农业科学院热带作物生物技术研究所, 热带作物生物技术国家重点实验室 ,海南海口 571101;2. 华南农业大学食品学院生物工程系, 广州 510640;3. 福建省莆田市农业科学研究所 ,莆田 351100)
Author(s):
Zhang Xiuchun 1 　 Peng Ming 1 　 Wu Kunxin 1 　 Guo Liqiong 1; 2 　 Lin Junyang 　 Lin Junfang1; 2
(1. State Key Laboratory of Tropical Crop Biotechnology , Chinese Academy of Tropical AgriculturalSciences, Haikou 571101 ;2 Department of Bioengineering , Collegeof Food Sciences , South ChinaAgricultural University , Guangzhou 510640;3 Putian City’s Institute ofAgricultural Sciences , Putian 351100)
关键词:
双 T - DNA ; 无标记基因 ; 大豆
Keywords:
Double T - DNA ; Marker-free ; Soybean
DOI:
10.11861/j.issn.1000-9841.2006.04.0369
文献标志码:
A
摘要:
利用 PCR 和 Southern 杂交检测了 161 株转 △6- 脂肪酸脱氢酶基因(D6D)的 T 1 代植株,初步筛选出只含有功能基因(D6D)而不含有筛选标记基因(bar)的转基因大豆植株 9 株 ,为获得富含 γ-亚麻酸但不含选择标记基因的转基因大豆奠定了基础, 并为转基因大豆的安全性种植提供了保障 。同时对无标记基因的转基因植株进行了叶片涂抹除草剂验证, 探讨了叶片涂抹除草剂结合目的基因 PCR检测筛选无选择标记转基因植株的可行性 。
Abstract:
T ransgenic crops have developed in commercialized scale in some countries, but their biosafetyhave attracted public concerns in the world at the same time. Therefore , it is becoming more and more im-portant to eliminate the selective genes because of their negative functions, such as antibiotics resistanceand herbicide resistance. 161 T 1 transgenic soybean of pudou 8008 containing Δ6- fatty acid desaturasegene and bar selection gene , which were obtained by biolistic particle co-transformation, were analyzed byPCR and southern blot in this study , and 9 plants only with the target gene were identiffed finally. Moreo-ver , leaf painting of herbicide was applied to confirm the marker free transgenic plants. It could be more ef-ficient to combine leaf painting and PCR, Southern blot to select bar free transgenic soybean.

参考文献/References:

1　 Komari T , Hiei Y , Saito Y, et al. Vectors carrying two separateT - DNAs for co - transformation of higher plants mediated byAgrobacterium tumefaciens and segregation of transformants freefrom selection markers. Plant J, 1996, 10(1):165 - 174.

2　 Holger P. Removing selectable marker genes:taking shortcut[ J] . T rends in Plant Sci, 2000, 5(7):273- 274.
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4　 Xing AQ , ZhangYZ , Shirley S , et al. T he use of the two T -DNA binary system to derive marker - free transgenic soybeans[ J] . In Vitro Cell Dev Biol - Plant , 2000, 36:456- 463.
5　 张秀春, 林俊芳, 郭丽琼. Δ 6 - 脂肪酸脱氢酶基因克隆及其共转化表达载体的构建[ J] . 热带作物学报, 2004, 25(4):63 - 67.
6　 张秀春, 郭丽琼, 吴坤鑫, 等. 双 T - DNA 表达载体转化大豆的研究[ J] . 大豆科学, 2005, 24(4):291- 295.
7　 张新梅、徐惠君、杜丽璞、等. 共转化法剔除转基因小麦中的 bar基因[ J] . 作物学报, 2004, 30(1):26- 30.
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备注/Memo

基金项目:广东省科技攻关项目(2005B20101009), 国家自然科学基金(30371000)资助。

作者简介:张秀春(1972 -), 女, 副研究员, 博士,主要从事植物生物技术研究。
通讯作者:林俊芳, E - mail:linjf@scau. edu. cn

更新日期/Last Update: 2016-12-16