Molecular detection method and kit for rice photosensitive genic male sterility lines

Abstract

The invention discloses molecular detection method and kit for rice photosensitive genic male sterility lines. The molecular detection method includes extraction of genome DNA, PCR (polymerase chain reaction) amplification, and enzyme digestion analysis of amplification products. Two pairs of primers and two endonucleases are designed according to a PMS12-1 sequence, and the primers are used for enzyme digestion analysis on the amplification products by PCR so as to determine SNP (single nucleotide polymorphism) type of the photosensitive gene. Therefore, whether the photosensitive genic male sterility gene or fertile gene is contained or not is identified, homozygote and heterozygote of the PMS12-1 gene can be further distinguished, the rice photosensitive genic male sterility lines are identified fast, simply and conveniently, and selection in fields is guided to accelerate breeding process.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method and a kit for detecting photosensitive genetic sterile lines by using two pairs of photosensitive gene primers and two kinds of endonucleases. Background technique [0002] The promotion and application of hybrid rice has greatly contributed to the increase in global rice production. Photosensitive GMS (PGMS) is the core part of two-line crop hybrids. Hybrid rice seed production using photosensitive genic male sterile lines has been very successfully developed and widely used. Therefore, the identification of sterile lines is very important in hybrid rice breeding. [0003] Research on the molecular mechanism of rice fertility transition controlled by light and temperature conditions has made great progress. Most eukaryotic genome sequences are transcribed into long non-coding RNAs (lncRNAs). Among them, long-day-specific male fertility-associated RNA (LDMAR) regulates PSM...

AI Technical Summary

Problems solved by technology

However, this method has the following disadvantages: the rice growth period is too long and limited by time, the fertility division is vague and difficult to judge

Sequencing can directly measure the base substitutions of all mutation sites in the DNA sequence, but this method requires expensive instruments and long steps, and the cost is high; the SSCP method is relatively mature, but the operation is cumbersome, time-consuming, and the result is easy. Causes misjudgment; PCR-RFLP method requires that the polymorphic site to be detected is related to a certain restriction site

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