[1]高嵩,何欢,吴楠,等.抗虫基因cry1Ab13 在大豆中的遗传转化及抗虫性鉴定[J].大豆科学,2016,35(04):550-556.[doi:10.11861/j.issn.1000-9841.2016.04.0550]
　GAO Song,HE Huan,WU Nan,et al.Identification on Insect-resistant Gene of cry1Ab13 Transformation into Glycinemax L. Merr.[J].Soybean Science,2016,35(04):550-556.[doi:10.11861/j.issn.1000-9841.2016.04.0550]
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抗虫基因cry1Ab13 在大豆中的遗传转化及抗虫性鉴定

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年04期 页码: 550-556 栏目: 出版日期: 2016-07-20
Title:
Identification on Insect-resistant Gene of cry1Ab13 Transformation into Glycinemax L. Merr.
作者:
高嵩1; 何欢1; 吴楠1; 韩丹1; 曲静1; 张君1; 张凤晶2; 王丕武1
1. 吉林农业大学生物技术中心，吉林长春，130118;?
2. 长春八一医院，吉林长春，130061
Author(s):
GAO Song1; HE Huan1; WU Nan1; HAN Dan1; QU Jing1; ZHANG Jun1; ZHANG Feng-jing2; WANG Pi-wu1
1. Biotechnology Center of Jilin Agricultural University，Changchun 130118，China;?
2. Changchun Bayi Hospital，Changchun 130061，China
关键词:
大豆;  cry1Ab13 基因;  植物表达载体;  抗虫
Keywords:
Soybean;  Cry1Ab13;  Plant expression vector;  Insect-resistant
分类号:
S332. 1
DOI:
10.11861/j.issn.1000-9841.2016.04.0550
文献标志码:
A
摘要:
cry1Ab13 基因是抗虫基因，编码的杀虫晶体蛋白对鳞翅目害虫具有一定的毒杀作用，构建了植物表达载体pCambia3300-35S-cry1Ab13，通过花粉管通道法将该载体转入大豆品种吉农28 中，经PCＲ 扩增检测得到15 株T1代阳性植株。Southern blotting 分析显示有5 株出现杂交信号，并以单拷贝形式整合到大豆基因组中。荧光定量PCＲ 测定结果表明: cry1Ab13 基因在转化植株的叶、茎秆中均有表达，每株表达量各不相同，在叶片部位表达量相对较高，最高为6. 7，最低为2. 5; 在茎部表达量最高为0. 76，最低为0. 31。对T1代阳性植株籽粒，采用圆盘分隔法接入大豆食心虫幼虫，进行初步抗虫试验，结果显示转化植株抗虫效果明显，但后代稳定性还需进一步研究。
Abstract:
The cry1Ab13 gene used in this experiment is the insect resistant gene，it encodes insecticidal crystal protein andhas certain toxic killing effect on Lepidoptera pests. The plant expression vector of pCambia3300-35S-cry1Ab13 was constructed.The transgenic plants were obtained by pollen-tube pathway transformation of soybean JN28. 15 transformed plants in T1generation were tested by PCＲ，while Southern blotting indicated that only 5 of them showed hybridization bands，and thefunctional fragment was integrated into the soybean genome by single copy. The result of quantitative real-time PCＲ showedthat the cry1Ab13 gene was expressed in soybean leaves and stems，and each plant expression was different， the relatively highexpression was in leaves， the highest was 6. 7，the lowest was 2. 5，while it was 0. 76 and 0. 31 in the stems. The seeds ofPCＲ positive plants in T1 generation and newly hatched larvaes were introduced，and initial insect resistant assay was made bydisk method. Ｒesult suggested that transgenic plant had an obvious resistant effect. Further investigations on the genetic stabilityof cry1Ab13 in transgenic plants and generations are to be carried out.

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备注/Memo

基金项目: 转基因生物新品种培育重大专项( 2014ZX08004-004) 。第一作者简介: 高嵩( 1989-) ，女，硕士，从事生物技术与作物遗传育种研究。E-mail: gaosong2425@ icloud. com。通讯作者: 王丕武( 1958-) ，男，博士，教授，主要从事大豆遗传育种研究。E-mail: peiwuw@163. com。

更新日期/Last Update: 2016-08-22