Quick detection method of exogenous herbicide resistance gene flow on soybean plant

Abstract

The invention discloses a quick detection method of exogenous herbicide resistance gene flow on soybean plant. The method comprises the following operation steps: (1) cutting off leaves of a to-be-detected soybean plant, positive reference plants and negative reference plants at the leaf stalks, wherein the number of each of the negative reference material and the positive reference material is two; (2) inserting the obtained leaves into centrifugal tubes to perform water planting; (3) placing the centrifugal tubes filled with the water-planting blades in a greenhouse to culture, spraying herbicide on the to-be-detected water-planting leave, the negative reference material CK3 and the positive reference material CK4, wherein the negative reference material CK1 and the positive reference material CK2 blades are blank control and free from spraying herbicide; and (4) observing after 5-7 days. The method provided by the invention is simple and quick in detection and low in cost, and a detection result is highly consistent with the traditional PCR detection and protein test strip detection result, and free from significant difference. The quick detection method disclosed by the invention can be used as a high-throughput method for important genetic or breeding material gene flow screening. And the method can provide technical support for protecting the import genetic germ plasm purity and genetic integrity.

Description

technical field [0001] The invention belongs to the technical field of gene drift detection, and in particular relates to a rapid detection method for exogenous herbicide-resistant gene drift of soybean plants. Background technique [0002] Gene drift is an important process of biological evolution and a common natural phenomenon, but if the exogenously introduced genes are released into the ecological environment through gene drift, it may lead to serious genetic and ecological consequences (Guo Lilei et al., 2019). In recent years, the research and development of genetically modified soybeans has been rapid, especially the promotion of CP4-epsps genetically modified glyphosate herbicide-resistant varieties worldwide, which has greatly improved the efficiency of weeding in farmland and achieved huge economic and social benefits (Pu Hongshan, 2019) , but the flow of resistance genes in transgenic soybeans has attracted more and more attention (Zhou Bo, 2006). [0003] Norma...

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Problems solved by technology

[0004] The identification of herbicide spraying can quickly verify the existence of exogenous herbicide tolerance genes in plants, but due to the killing properties of herbicides, for important genetic materials, such as breeding progeny and genetic sterile recurrent selection populations, identification is not possible. The main purpose of source gene drift is to keep the uncontaminated test materials as complete as possible, and it is not possible to simply identify and verify individual plants with gene drift through herbicide spraying

[0005] The PCR detection method is a national standard method for detecting exogenous herbicide-resistant genes. The result is accurate, but it takes a long time and costs a lot. It requires professional laboratories and equipment, and requires high requirements for operators.

[0006] In addition, in corn research, the protein test strip method is often used to detect the herbicide-resistant CP4-epsps gene. The cost of glyphosate herbicide CP4-epsps gene detection is relatively low. If there is no related protein test strip product development for other herbicide resistance genes, the protein test strip method will not be available, and the protein test strip method is only applicable For the rapid detection of a small number of samples in the field, the cost of high-throughput detection is relatively high

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