Molecular Identification and Chromosomal Mapping of a Low Toxic Bt Gene in Upland Cotton

doi:10.7668/hbnxb.20193244

Abstract

Abstract:

In order to analyze a transgenic event of an upland cotton germplasm strain with low expression of a Bt gene,specific primers were used to identify the Bt gene.The PCR amplification result showed a fragment length of the Bt gene was 310 bp,which conformed characteristic primer fragment length to Bt gene of the United States.Using the germplasm strain to cross with island cotton Shengli No.1 showed that the Bt gene was dominant in F₁,and Bt-harboring plants to non-harboring plants displayed a 3∶1 segregation ratio in F₂ population.The result proved that the Bt gene was a quality trait controlled by a pair of dominant gene,which meant the foreign Bt gene was inserted into upland cotton as a locus.Furthermore,F₂ population and SSR markers were used in the gene mapping.The result revealed that the Bt gene was mapped on the 10th chromosome of cotton,and 14 pairs of primers were linked to the target gene,such as NAU5166,NAU3574,NAU456,BNL256,cgr6745,cgr5406,cgr6546,NAU7110,HAU3201,BNL1665,dPL0468,NAU5316,BNL2960,NAU3122.The Bt gene was located between the molecular markers NAU7110 and HAU3201,and its genetic distance was 0.9,4.4 cM,respectively.

Key words: Upland cotton, Low-toxic Bt gene, Molecular identification, Gene mapping

CHEN Xusheng, ZHAO Liang, DI Jiachun. Molecular Identification and Chromosomal Mapping of a Low Toxic Bt Gene in Upland Cotton[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(6): 103-107. doi: 10.7668/hbnxb.20193244.

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Figures/Tables 4

Fig.1 The origin of Bt gene in germplasm strain WG-20 M.DNA Marker;1.GK-19;2.WG-20;3.Distilled water;4.Simian No.3.

Fig.2 The characteristic bands of Bt gene in parents and F₁,F₂ populations M.Marker;P1.WG-20;P2.Shengli No.1;F₁.WG-20×Shengli No.1;1—36.Partial plants of F₂ population.

Tab.1 Isolation of Bt gene from the offspring of WG-20 crossing Shengli No.1

世代 Generation	亲本及组合 Parents and combinations	有Bt基因个体数 Number of harboring Bt individuals	无Bt基因个体数 Number of non-harboring Bt individuals	理论比例χ²值 F₂ 3∶1 Theoretical ratio of χ² value
P1	WG-20	43	0	—
P2	胜利1号	0	45	—
F₁	P1×P2	48	0	—
F₂	F₁自交	143	35	2.70

Fig.3 Genetic linkage map of low-toxic Bt gene in Chr.10

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