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作物学报 ›› 2019, Vol. 45 ›› Issue (9): 1440-1445.doi: 10.3724/SP.J.1006.2019.84167

• 研究简报 • 上一篇    下一篇

2个抗虫棉的外源Bt基因分子鉴定及其染色体定位

周向阳1,赵亮2,狄佳春2,陈旭升2,*()   

  1. 1 南京农业大学农学院, 江苏南京 210095
    2 江苏省农业科学院经济作物研究所, 江苏南京 210014
  • 收稿日期:2018-12-12 接受日期:2019-04-15 出版日期:2019-09-12 网络出版日期:2019-05-14
  • 通讯作者: 陈旭升
  • 作者简介:E-mail: xiangyzhou@126.com
  • 基金资助:
    本研究由国家转基因生物新品种培育科技重大专项子课题资助(2016ZX08005001-008);本研究由国家转基因生物新品种培育科技重大专项子课题资助(2016ZX08005-005)

Molecular identification and chromosomal mapping of exogenous Bt gene in two insect-resistant cotton varieties

ZHOU Xiang-Yang1,ZHAO Liang2,DI Jia-Chun2,CHEN Xu-Sheng2,*()   

  1. 1 Agronomy College, Nanjing Agricultural University, Nanjing 210095, Jiangsu, China
    2 Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu, China
  • Received:2018-12-12 Accepted:2019-04-15 Published:2019-09-12 Published online:2019-05-14
  • Contact: Xu-Sheng CHEN
  • Supported by:
    This study was supported the National Major Project for Developing New GM Crops(2016ZX08005001-008);This study was supported the National Major Project for Developing New GM Crops(2016ZX08005-005)

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摘要：

以中美2个抗虫棉品种GK19与33B为试验材料, 利用检测中美Bt基因的特异性引物, 分别对抗虫棉亲本GK19和33B进行PCR扩增, 并通过SSR分子标记技术对其Bt基因进行分子鉴定与染色体定位, 旨在从外源基因转化事件的视角探究中美转基因抗虫棉差异的分子基础。结果表明, GK19为中国转Bt基因抗虫棉, 33B为美国转Bt基因抗虫棉; GK19的Bt基因被定位在棉花Chr.20上, 共16对SSR多态性标记与其Bt基因连锁, 两侧的分子标记为NAU3907和NAU2579, 其遗传距离分别为2.4 cM和1.5 cM; 33B的Bt基因被定位在棉花Chr.26上, 共20对SSR多态性标记与Bt基因连锁, 目标Bt基因位于标记NAU460和dc40260之间, 其遗传距离分别为3.6 cM和2.0 cM。以上结果表明GK19和33B属于不同的遗传转化事件。

关键词: 抗虫棉, Bt基因, PCR检测, 染色体定位, 转基因事件

Abstract:

Molecular identification and chromosomal localization of Bt genes in two insect-resistant cotton varieties GK19 from China and 33B from the United States were conducted to explore the molecular basis of the differences between the two transgenic cotton varieties in perspective of exogenous gene transformation events. GK19 was a domestic Bt transgenic insect-resistant cotton variety, its Bt gene was located on cotton Chr.20, and linked to a total of 16 pairs of SSR polymorphic markers, flanking sides were markers NAU3907 and NAU2579 with genetic distance of 2.4 cM and 1.5 cM, respectively. The Bt gene in 33B was located on cotton Chr.26, with a total of 20 pairs of SSR polymorphic markers linking to the Bt gene, the target Bt gene was located between markers NAU460 and dc40260, with genetic distance of 3.6 cM and 2.0 cM, respectively. This study revealed that GK19 and 33B were from different transgenic events.

Key words: insect-resistant cotton, exogenous Bt Genes, PCR detection, chromosomal localization, transgenic event

图1

Bt基因类型鉴定 M: DNA marker; 1: GK19; 2: 33B; 3: 清水; 4: 泗棉3号。"

图2

中国产Bt基因的PCR扩增 M: DNA marker; P1: 胜利1号; P2: GK19; F1为杂交一代, 1至33为F2代分离群体的部分单株。"

表1

GK19和胜利1号杂交后代Bt基因的分离情况"

世代
Generation		 亲本及组合
Parent and combination		 含Bt基因个体数
Number of individuals
with Bt genes		 不含Bt基因个体数
Number of individuals
without Bt gene		 χ2值
χ2 value
P1 V-1 0 83 —
P2 GK19 91 0 —
F1 P2×P1 80 0 —
F2 F1代自交 Self-crossing F1 133 51 0.72

图3

美国Bt基因的PCR扩增 M: DNA marker; P1: 胜利1号; P2: 33B; F1为杂交一代, 1至33为F2代分离群体的部分单株。"

表2

33B×胜利1号杂交后代Bt基因的分离情况"

世代
Generation		 亲本及组合
Parent and combination		 含Bt基因单株数
Number of individuals
with Bt gene		 不含Bt基因单株数
Number of individuals
without Bt gene		 χ2值
χ2 value
P1 V-1 0 84 —
P2 33B 90 0 —
F1 P2×P1 76 0 —
F2 F1代自交 Self-crossing F1 148 37 2.47

图4

抗虫棉GK19的外源Bt基因连锁遗传图谱"

图5

抗虫亲本33B的外源Bt基因连锁遗传图谱"

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