A molecular detection method for rapid and accurate identification of weedy rice and cultivated rice

Abstract

The invention discloses a molecular detection method for rapidly and accurately identifying weedy rice and cultivated rice, and belongs to the technical field of biology. The method includes the following steps: extracting a rice genomic DNA sample; performing nested PCR amplification on the rice genomic DNA sample; taking a PCR product as a substrate, and performing enzyme digestion reaction by using corresponding restriction enzyme; and detecting an enzyme-digested product through agarose gel electrophoresis to identify weedy rice and cultivated rice. The method can perform identification inthe seedling stage of rice, so that the purpose of early discovery and early weedy rice control can be achieved. The method adopts a nested PCR method to perform amplification on SNP sites, so that the specificity and sensitivity of amplification reaction can be guaranteed, and the obtained PCR product can be directly used for the enzyme digestion reaction of the restriction enzyme; and the wholeprocess does not exceed 6 hours, and needed instruments and reagents are easy to obtain and cheap in price, so that the method is simple in operation. The method designs three pairs of specific primers for three SNP sites, and adopts triple certification of detection results to guarantee the accuracy of the results.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a molecular detection method for quickly and accurately identifying weedy rice and cultivated rice. Background technique [0002] Weedy rice is a kind of Oryza plant that grows in rice fields and grows with cultivated rice, and is between wild rice and cultivated rice in form. Weedy rice has a strong ability to adapt to the environment, especially in paddy fields, seriously endangering rice production. At present, weedy rice has been found in major rice producing areas in the world. In the United States, due to long-term competition between weedy rice and rice, rice production has been reduced by as much as 80%. Compared with cultivated rice, weedy rice has the characteristics of early heading and strong grain shattering. Therefore, under the same environmental conditions, weedy rice matures and shatters earlier, causing the base number of weedy rice in the field to expand year by y...

AI Technical Summary

Problems solved by technology

However, traditional weedy rice identification methods are mainly based on the analysis of biological traits such as plant type, number of tillers, and grain type. Only in the late tillering stage will weedy rice show higher plant height and more tiller numbers than cultivated rice. , the leaf sheath, ligule, glume, seed coat and other parts of the plant often show phenotypic characteristics such as reddish brown, and the glume color of some weedy rice is very similar to that of cultivated rice

Therefore, the traditional phenotypic identification method has the shortcomings of unclear identification standards, large individual differences, and late identification time, which requires a method that can quickly and accurately distinguish weedy rice from cultivated rice at the seed germination stage or seedling stage , to radically reduce the amount of weedy rice in paddy fields

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