Molecular marker method for identifying indica type rice and japonica rice by using rice grain

Abstract

The invention belongs to the technical field of biotype identification, in particular to a method for identifying indica type rice and japonica rice using rice grain by using a rice grain (rice) and inserting or deleting (InDel) a molecular marker. The method comprises the following steps of: extracting DNA from the rice grain, and comparing full-genome DNA sequences of indica type rice 93-11 andjaponica rice Nipponbare to obtain 40 pairs of specific InDel primers; and performing fragment amplification, electrophoretic separation and electrophoresis pattern analysis on the extracted DNA the in rice seed to identify the characteristics of the indica type rice and japonica rice of a rice sample. The method concretely comprises the following steps of: taking the DNA extracted from the rice grain as a template, and analyzing and counting molecular fingerprint patterns obtained on the basis of a polymerase chain reaction and agarose electrophoresis by using combination of the 40 pairs of specific InDel primers; and determining the characteristics of the indica type rice and japonica rice of the tested rice seed (sample) according to gene frequency (Fi or Fj) calculated by 93-11 genotype and japonica rice Nipponbare genotype molecular fingerprint on 40 InDel loci of the sample. The result can be obtained by only detecting the sample of one grain of seed, and the method is convenient and rapid, accurate in identification, and has good popularization and application prospect.

Description

technical field [0001] The invention belongs to the technical field of biological type identification, and specifically relates to a method for extracting DNA from a rice seed (or rice) and identifying its indica and japonica characteristics. technical background [0002] Cultivated rice ( Oryza sativa L.) is one of the three important food crops in China, and occupies a pivotal position in my country's agricultural production and economic development. Cultivated rice originated in my country, has a cultivation history of more than 8,000 years, and is currently widely planted all over the world. During the cultivation and domestication of cultivated rice, significant genetic differentiation between indica and japonica occurred due to adaptation to different ecological environments. There has been obvious reproductive isolation between typical indica rice and typical japonica rice, so indica and japonica rice are usually divided into two subspecies: indica subspecies ( Or...

AI Technical Summary

Problems solved by technology

The biggest disadvantage of this method: 1) The detection is inaccurate because the morphological traits are greatly affected by the environment; 2) The rice samples to be tested must be planted for morphological measurement, and it usually takes 3-5 months from sowing to maturity, which takes a long time ; 3) Morphological measurement requires a certain sample population, so more rice samples are required; it greatly affects the efficiency of identification of indica and japonica rice

However, the extraction of rice DNA is usually carried out with fresh leaves, and the rice seeds need to be germinated to form seedlings, and the DNA can be extracted after 1 week or even 10 days, which takes a long time

DNA cannot be extracted from seeds that cannot germinate

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