PCR identification method of soybean rhizobium strain 5873, and primers and application of method

Abstract

The invention relates to a PCR identification method of soybean rhizobium strain 5873, and primers and application of the method, and belongs to the technical field of strain identification. The method comprises the following steps: using the liquid after lysis, boiling water bath and freeze-thaw of the genomic DNA of strains to be tested or pure culture liquid of the strains or a root nodule crushed extract as templates, and performing PCR amplification using primers 4-4; performing electrophoresis of the PCR products, and identifying that the samples without bands around 355 bp are not the soybean rhizobium strain 5873; continuing PCR amplification of the strains to be test by using a primer Q1, when bands around 355 bp exist; and performing electrophoresis of the above products, and identifying that the samples with bands around 218 bp are the soybean rhizobium strain 5873. According to the present invention, the method has the following advantages that the soybean rhizobium strain5873 can be distinguished simply and quickly in terms of strain identification and evaluation, and bacterial bodies or root nodules can be directly used as templates, so that the tedious steps such asseparation, culture and DNA extraction and identification root nodules are omitted.

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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