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Primers for detecting exogenous genes in transgenic rice TT51-1 derivative line and method thereof

A technology for transgenic rice and derived lines, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. In order to improve the breeding efficiency and high-efficiency detection effect

Pending Publication Date: 2020-02-14
INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI
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Problems solved by technology

The molecular markers currently used to detect cry1Ab / cry1Ac genes are mainly specific dominant markers designed according to the sequence of imported foreign gene fragments. This marker can only detect the presence or absence of the target gene, and cannot distinguish heterozygous genotypes from homozygous genotypes. , the genotype needs to be verified in the next generation, so the detection efficiency of dominant markers is low and time-consuming

Method used

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  • Primers for detecting exogenous genes in transgenic rice TT51-1 derivative line and method thereof
  • Primers for detecting exogenous genes in transgenic rice TT51-1 derivative line and method thereof
  • Primers for detecting exogenous genes in transgenic rice TT51-1 derivative line and method thereof

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Embodiment Construction

[0019] The present invention will be further described below in combination with specific embodiments.

[0020] A method for detecting exogenous genes in transgenic rice TT51-1 derivative lines. This example uses the cry1ab / cry1ac gene homozygous line TT51-1 and non-transgenic rice R650, wherein the cry1ab / cry1ac gene homozygous line TT51- 1Provided by the State Key Laboratory of Crop Genetic Improvement / National Plant Gene Research Center (Wuhan), Huazhong Agricultural University, non-transgenic rice R650 is a rice restorer line bred by the Grain Crops Research Institute of the Hubei Academy of Agricultural Sciences. Crossbreeding non-transgenic rice R650 female parent and TT51-1 male parent to obtain cry1Ab / cry1Ac gene heterozygous F 1 Generation, F 1 Generation and R650 backcross to obtain BC 2 f 1 In the next generation, the positive individual plants detected by molecular markers continued to be backcrossed to obtain BC 3 f 1 Finally, according to the results of mole...

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Abstract

The invention discloses primers for detecting exogenous genes in a transgenic rice TT51-1 derivative line and a method thereof. A forward primer P1, a forward primer P3 and a reverse primer P2 are designed for an exogenous gene cry1Ab / cry1Ac in the transgenic rice TT51-1 derivative line, during detection, the three primers are adopted to perform multiplex PCR amplification on genomic DNA of TT51-1and progeny materials derived from TT51-1 in a same PCR system, and then the banding pattern of an amplification product is detected. According to the design, simple and efficient detection of existence and homozygosity of exogenous genes is realized.

Description

technical field [0001] The invention belongs to the field of plant breeding detection, and in particular relates to a primer and a method for detecting exogenous genes in transgenic rice TT51-1 derivative lines. Background technique [0002] Rice leaf roller, rice stem borer and rice stem borer are the main pests of rice, and their damage will cause leaves to wither, dead seedlings, white ears, and a large increase in shriveled grains, seriously affecting rice yield. Rice transgenic for Bt toxin protein has good resistance to rice leaf roller, rice stem borer and rice stem borer, planting Bt rice can effectively avoid the damage of rice leaf roller, rice stem borer and rice stem borer to rice , and can reduce the use of pesticides and reduce environmental pollution. Rice scientists have bred a series of rice with Bt genes (Tu et al., 2000; Ye et al., 2001; Chen et al., 2005; Tang et al., 2006), and these Bt rice were used for breeding new stem borer-resistant rice. Varieti...

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 万丙良査中萍殷得所杜雪树李进波夏明元戚华雄
Owner INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI
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