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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (9): 1440-1445.doi: 10.3724/SP.J.1006.2019.84167

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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 Online:2019-09-12 Published:2019-05-14
  • Contact: Xu-Sheng CHEN E-mail:njcxs@126.com
  • 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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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

Fig. 1

Identification of exogenous Bt genes M: DNA marker; 1: GK19; 2: 33B; 3: Water; 4: Simian 3."

Fig. 2

PCR amplification of Bt gene from China M: DNA marker; P1: V-1; P2: GK19; F1 for hybrid generation, 1 to 33 for partial plants in F2 segregation population."

Table 1

Bt gene separation in hybrid offspring of GK19 crossing V-1"

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

Fig. 3

PCR amplification of Bt gene from USA M: DNA marker; P1: V-1; P2 : 33B; F1 for hybrid generation, 1 to 33 for partial plants in F2 segregation population."

Table 2

Bt gene separation in hybrid offspring of 33B crossing V-1"

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

Fig. 4

Exogenous Bt gene linkage map of insect-resistant cotton GK19"

Fig. 5

Exogenous Bt gene linkage map of insect-resistant cotton 33B"

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