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

A technology of japonica and japonica rice, which is applied in the field of identifying the characteristics of indica and japonica rice, which can solve the problems of inaccurate detection, affecting the identification efficiency of indica and japonica rice, and extraction after 1 week or even 10 days.

Inactive Publication Date: 2013-06-12
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The DNA of each rice sample obtained by the method of rapid DNA extraction from one grain of rice was used as a template (see figure 1 ), utilizing a pair of InDel molecular markers (see figure 2 ) combined with agarose gel electrophoresis to identify the indica and japonica characteristics of 17 rice varieties.

[0055] Table 2. Experimental materials used for identification in Example 1

[0056] serial number rice variety source Indica-japonica gene frequency (Fi / Fj) (%) InDel identification results 1 Japan Sunny Japan 0 / 100 Typical japonica rice 2 Nanjing No. 11 Yangzhou 100 / 0 Typical Indica 3 93-11 China 100 / 0 Typical Indica 4 Minghui 86 Fuzhou 98 / 2 Typical Indica 5 Hubei early Jiangxi 99 / 1 Typical Indica 6 Red Sticky Valley Guizhou 98 / 2 Typical Indica 7 Pong Qiu Nuo Anhui 93 / 7 Typical Indica 8 Nippon Haruna 93-11 Hybrid F2 Wuhan 54 / 46 intermediate type 9 ...

Embodiment 2

[0086] The DNA of each rice sample obtained by the method of rapid DNA extraction from one grain of rice was used as a template (see figure 1 ), using three pairs of InDel molecular markers to more efficiently identify the indica and japonica characteristics of unknown rice varieties (see image 3 ). The experimental materials are the same as in Example 1 (see Table 3).

[0087]The DNA extraction method of the rice sample, the reaction system used in the PCR reaction, the procedure of the PCR reaction, the electrophoresis detection, and the genotype reading and analysis methods of different rice samples are the same as in Example 1.

[0088] image 3 The results showed that three pairs of InDel primers were used for PCR amplification of different rice varieties, and the PCR products were detected by electrophoresis at the same time, the PCR products of these three pairs of InDel primers could be clearly distinguished, and at the same time, different rice DNA The samples we...

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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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12N15/11C12Q1/68
Inventor 卢宝荣蔡星星刘苹
Owner FUDAN UNIV