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Auxiliary method for identifying wheat with different thousand grain weight properties, and special primer thereof

An auxiliary identification and thousand-grain weight technology, applied in the direction of botanical equipment and methods, biochemical equipment and methods, applications, etc., to achieve the effects of avoiding influence, optimizing hybrid combination, and speeding up breeding

Inactive Publication Date: 2012-07-25
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the cloning of the GIF gene in wheat

Method used

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  • Auxiliary method for identifying wheat with different thousand grain weight properties, and special primer thereof
  • Auxiliary method for identifying wheat with different thousand grain weight properties, and special primer thereof
  • Auxiliary method for identifying wheat with different thousand grain weight properties, and special primer thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Obtaining of Proteins Related to Wheat Grain Weight and Their Encoding Genes

[0031] 1. Acquisition of TaCwi-A1a protein and its coding gene

[0032] The TaCwi-A1a protein coding gene was obtained by combining electronic cloning with conventional PCR amplification. According to the rice grain weight-related gene GIF gene (GenBank Accession Number: EU095553), BLAST search was performed in the wheat EST database, and the screened ESTs were spliced ​​respectively, and then the full-length CWI gene was obtained by electronic splicing according to CK206103, CD927722, CJ673133, and D927242 , using Primer Primer5 software to design 3 pairs of primers C1, C2, C3, to obtain the true CWI gene sequence from the genomic DNA of the wheat cultivar Soybean.

[0033] The PCR reaction was carried out on a MJ Research PTC-200 PCR instrument using wheat genomic DNA as a template. The 20 μl PCR reaction system was: 50 ng of template DNA, 1 U of Taq enzyme (Beijing Tiangen Bioc...

Embodiment 2

[0042] Embodiment 2, correlation analysis of wheat grain weight character and genotype

[0043] 1. Design of functional marker primers

[0044] According to sequence 2 and sequence 4 in the sequence listing, two groups of primers CWI21 (primer pair B; P1 / P2) and CWI22 (primer pair A; P3 / P2) are designed as follows:

[0045] P1 (upstream primer): 5'-GTGGTGATGAGTTCATGGTTAAG-3' (SEQ ID NO: 5);

[0046] P3 (upstream primer): 5'-GGTGATGAGTTCATGGTTAAT-3' (SEQ ID NO: 6)

[0047] P2 (downstream primer): 5'-AGAAGCCCAACATTAAATCAAC-3' (SEQ ID NO: 7).

[0048] With primer pair B, the DNA fragment of 404bp in the nucleotide shown in sequence 4 in the sequence listing can be amplified, and the DNA fragment in the nucleotide shown in sequence 2 in the sequence listing cannot be amplified; With primer pair A, the sequence can be amplified The 402bp DNA fragment in the nucleotides shown in sequence 2 in the list cannot amplify the DNA fragment in the nucleotides shown in sequence 4 in the s...

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Abstract

The invention discloses an auxiliary method for identifying wheat with different thousand grain weight properties, and a special primer thereof. The method comprises the following steps of: using a wheat gene group DNA as a template, and carrying out PCR enlargement by a primer pair A and a primer pair B, wherein if the enlargement product of the primer pair A has a 402bp DNA segment, while the enlargement product of the primer pair B does not have the 402bp DNA segment, the gene type of the wheat to be tested is TaCwi-Ala; if the enlargement product of the primer pair B has the 402bp DNA segment, while the enlargement product of the primer pair A does not have the 402bp DNA segment, the gene type of the wheat to be tested is TaCwi-Alb; and the thousand grain weight of the wheat with the gene type of TaCwi-Ala is greater than that of the wheat with the gene type of TaCwi-Alb in statistics. The invention can be used for culturing fine wheat varieties with high thousand grain weight, and avoid sieving the wheat varieties with low thousand grain weight. The wheat varieties can be sieved in the early period of growth, therefore, the hybridized combination is optimized, the breeding speed is accelerated, and the breeding cost is decreased.

Description

technical field [0001] The invention relates to a method for assisting identification of wheat with different thousand-grain weight traits and special primers thereof. Background technique [0002] Wheat is one of the most important food crops in the world. The calories and protein it provides account for more than 20% of human nutrition. More than 35% of the world's population uses wheat as their staple food. my country's wheat planting area, total output and consumption rank first in the world, and its output accounts for about 1 / 5 of my country's total grain output. [0003] Improving wheat yield is an important goal of wheat breeding. The yield per unit area of ​​wheat is composed of the number of ears per unit area, the number of grains per ear and the average grain weight. 1000-grain weight has the greatest direct contribution to yield among the yield components, and increasing the average grain weight per panicle is the key to improving the yield potential of variet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/29C07K14/415
Inventor 夏先春马冬云何中虎
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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