Identification method of soybean rhizobium strain 5873 pcr, its primers and application

Abstract

The invention relates to a PCR identification method for soybean rhizobium strain 5873, primers used therefor and applications, belonging to the technical field of bacterial strain identification. The method comprises: using the genomic DNA of the bacterial strain to be tested or the pure culture bacterial liquid of the bacterial strain or the root nodule crushing extract after cracking, boiling water bath, and freeze-thawed liquid as a template, and performing PCR amplification with primer 4-4; Perform electrophoresis on the PCR product, and if there is no band of about 355bp, it is not soybean rhizobium strain 5873; if there is a band of about 355bp, continue to use primer Q1 for PCR amplification of the strain to be tested; perform electrophoresis on the above product, and there is a band of about 218bp Soybean Rhizobium strain 5873. The present invention has the following advantages: In terms of strain identification and evaluation, soybean rhizobia strain 5873 can be easily and quickly distinguished, and bacteria or nodules can be directly used as templates, eliminating the cumbersome separation, cultivation, and DNA extraction and identification of nodules. link.

Description

technical field [0001] The invention belongs to the technical field of strain identification, and in particular relates to a PCR identification method of soybean rhizobium strain 5873, primers used therein and applications. Background technique [0002] With the adjustment of the country's agricultural strategy, the development of resource-saving and environment-friendly "two-type agriculture" has become the only way for the sustainable development of modern agriculture. Among them, microbial fertilizers including rhizobia, as new green inputs and priority biological products, can not only repair and improve soil, reduce weight and increase efficiency, but also perform outstandingly in improving crop quality and efficiency. The action plan of reducing one increase provides an important guarantee for the realization of the goal of "one control, two reductions and three basics". [0003] As an important functional strain in microbial fertilizer products, soybean rhizobia can ...

AI Technical Summary

Problems solved by technology

In view of the fact that similar species of soybean rhizobia have similar colony morphology, we usually rely on basic physiological and biochemical reactions in combination with conserved sequences such as 16S rRNA, atpD, glnII, recA, gyrB and rpoB genes to identify the species, and the results are usually also It can only be distinguished at the species level, but cannot be identified at the strain level, which is not conducive to the protection of intellectual property rights of the production strains

Taking bradyrhizobia as an example, there is no strain-level PCR rapid detection technology, especially B. japonicum E109 and B. japonicum USDA 6, which are commonly used in domestic and foreign research and industrial applications. T , at the genome-wide level, we only found 3 SNP differences, which are difficult to distinguish by conventional technical means

In addition, at the application level, the nodulation rate is an important indicator for evaluating the nodulation ability of rhizobia after inoculation. It is usually determined by BOX-PCR technology. Although this method can distinguish species or some strains, the process is cumbersome and complicated. poor repeatability

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