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Method for screening medium amylase content rice and primer special for method

An amylose content, rice technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of inability to separate medium amylose and high amylose genotypes, etc. Breeding cycle, short cycle, cost reduction effect

Inactive Publication Date: 2013-02-06
INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI
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  • Application Information

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Problems solved by technology

In previous studies, it was found that wxya A SNP site In1 in the first intron of the gene is an important factor affecting amylose, and a molecular marker has been developed, but the variation of this site can only distinguish the low amylose content genotype, and cannot Medium amylose and high amylose genotypes are separated, so there are great limitations in practical breeding

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  • Method for screening medium amylase content rice and primer special for method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Using the special primer Wx-Ex6 in the present invention, molecular detection was carried out on 200 micro-core collections of cultivated rice in China and 95 core collections of foreign introduced species, including 111 indica varieties and 89 japonica varieties.

[0019] DNA was extracted using conventional CTAB extraction method.

[0020] The PCR amplification system is: rice genomic DNA template 50ng, Taq DNA polymerase 1 U, 1.5μl 10×PCR buffer (100Mm Tris-HCl (pH8.3), 500Mm KCl), 25mM MgCl 2 0.9ul, 2.5 mmol each dNTPs 0.6ul, 0.2μM for each of the four primers, and finally use ddH 2 O Supplement the reaction system to 15 μl.

[0021] The PCR reaction conditions can be: first 95°C for 5min; then 95°C for 30sec, 55°C for 30sec, 72°C for 40sec, a total of 35 cycles; then 72°C for 5min.

[0022] Detection of PCR products: Load the PCR products on a 1.5% agarose gel to detect the band pattern of the PCR products.

[0023] Molecular results showed that there were 766b...

Embodiment 2

[0026] First plant the above-mentioned Jufeng A / / Tianfeng A / J521 F in the field 6There are 114 lines in total in the generation breeding group; J521 contains the pedigree of the medium-straight variety Lemont, while Jufeng and Tianfeng are both tall-straight varieties. The method of the invention and the provided molecular marker Wx-Ex6 are used for detection at the seedling stage. DNA extraction, PCR amplification system and PCR product detection PCR product detection methods are the same as in Example 1.

[0027] DNA was extracted from these 114 strains, and the above-mentioned detection process was carried out, such as figure 1 As shown, the leftmost band is the molecular weight control Marker DL2000, P1 is the band type of the positive parent Lemont, the band size is 1025 and 766bp; P2 is the band type of the negative parent Jufeng A, the band size is 1025 and 310bp; 1-20 are the 20 rice lines to be detected, and the ones showing the same band size as P1 are wxya in P...

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Abstract

The invention discloses a method for screening medium amylase content rice and a primer special for the method. The primer for assisting to screen the rice is four primers composed of nucleotide sequences of sequence WxMF, WxMR, WxHF and WxHR in a sequence table. The method for assisting to screen the rice comprises the following steps of: using a genomic DNA (Deoxyribonucleic Acid) of the rice to be detected to serve as a template, using the four primers composed of nucleotide sequences of sequence WxMF, WxMR, WxHF and WxHR in the sequence table to conduct PCR (Polymerase Chain Reaction) amplification, if an amplification product of the rice to be detected has 766bp stripes, the rice to be detected is candidate rice. The method for assisting to screen medium amylase content rice can be used for selecting and cultivating the medium amylase content rice, shortens breeding cycles of the rice, quickens breeding speed, reduces breeding cost, has the advantages of being simple in operation, low in cost and short in period, is applicable to popularization and application and provides a rapid method for selecting and cultivating medium amylase content rice varieties.

Description

technical field [0001] The invention relates to a method for assisting in screening rice with medium amylose content and special primers thereof. Background technique [0002] Rice is one of the main food crops in the world. The rice sown area in my country is 28-32 million hectares, accounting for 27% of the national grain sown area; the rice yield per unit area is 5.8-6.3 tons / ha, and the total rice output is 180-200 million tons, accounting for 39% of the total grain output (agricultural Ministry of Agriculture, 2007 China Agricultural Development Report, China Agricultural Press, 2008, pp6-11). Rice production plays a very important role in my country's agricultural production and rural economic development, and is also the basis for my country's food security, social stability and national economic development. [0003] With the improvement of living standards, people's requirements for rice are not only to satisfy food and clothing, but also put forward higher and hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 游艾青周雷陈志军马银花陈少愚杨国才李三和刘凯胡刚闸雯俊
Owner INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI