Method for identifying soybean male sterile cytoplasm through SNP marks of chloroplast DNA

Abstract

The invention provides a method for identifying soybean male sterile cytoplasm through SNP marks of chloroplast DNA. The method can also be applied to distinction of a soybean cytoplasmic male sterile line and a maintainer line containing normal cytoplasm. The method includes the following steps of extracting the seed genome DNA of the soybean cytoplasmic male sterile line and the seed genome DNA of the maintainer line to serve as amplification templates, selecting four pairs of the SNPs marks of the chloroplast DNA, and designing a pair of primers on the side wing sequences of each pair of SNPs respectively to conduct PCR amplification. The PCR products obtained through amplification conduct identification on the difference between the length of the enzyme-digested products of the sterile line and the length of the enzyme-digested products of the maintainer line through digestion of restriction enzymes. According to the method, the soybean male sterile cytoplasm and normal fertile cytoplasm can be rapidly and accurately identified. The method can also be used for detecting the maintainer line which contains fertile cytoplasm and is mixed in the sterile line containing sterile cytoplasm in the breeding process of the sterile line, and provides guarantees for the purity requirement in the production process of soybean sterile line seeds.

Description

technical field [0001] The invention provides a method for identifying soybean male sterile cytoplasm by SNP marker of chloroplast DNA, which is used for identifying soybean male sterile cytoplasm and normal fertile cytoplasm, and can also be used to distinguish soybean cytoplasmic male sterile lines from sterility during seed production A paternal maintainer line (containing fertile cytoplasm) mixed into a line (containing sterile cytoplasm) belongs to the technical field of molecular marker detection. Background technique [0002] Soybean cytoplasmic male sterility (cytoplasmic male sterility, CMS) "three lines" consists of a sterile line, a maintainer line and a restorer line. The carrier of the male sterile cytoplasm is the sterile line, while the carrier of the male fertile cytoplasm is the maintainer line. The Jilin Academy of Agricultural Sciences first bred soybean CMS "three lines". CMS is widely used in the utilization of crop heterosis. Its biggest advantage is t...

AI Technical Summary

Problems solved by technology

Such seeds are planted in production, which will inevitably lead to sterile plants in the production field and reduced production

[0004] The current molecular marker method for identifying soybean male sterile cytoplasm is based on the difference in the size of PCR amplified fragments of base deletions (indels) to distinguish soybean RN-type cytoplasmic sterile lines and maintainer lines, and the amplified fragments in the sterile lines and The size difference in the maintainer line is only 12bp, which must be distinguished by high-concentration 3% agarose gel electrophoresis, which may not be easy to distinguish in practice, and a single marker may affect the soybean male sterile cytoplasm and normal fertile cytoplasm The accuracy of identification, which in turn affects the commercial breeding of soybean CMS

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