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Development method and application of KASP molecular marker of waxy gene in corn

A waxy gene and molecular marker technology, which is applied in biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc., can solve problems such as low detection efficiency, aerosol pollution, and inappropriate molecular detection platforms. To achieve the effect of simple operation, promotion of corn waxy breeding, and short detection period

Inactive Publication Date: 2021-10-22
YANGZHOU UNIV
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Problems solved by technology

[0004]However, although the linkage markers of many of the above-mentioned corn starch-related genes can be used for molecular marker-assisted selection, most of them are RFLP markers, SSR markers, InDel markers or enzyme-cut markers , the detection efficiency is low, and it may also produce aerosols to pollute the environment, which is not suitable for high-throughput molecular detection platforms (Cheng Beijiu, Chen Feng, Zhu Suwen, etc. Molecular markers of maize ae gene and its obtaining method and application, CN101619362B [P].2011; Lei Wusheng. Application of specific PCR in detection of maize ae gene[D]. Gansu Agricultural University, 2005)

Method used

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  • Development method and application of KASP molecular marker of waxy gene in corn
  • Development method and application of KASP molecular marker of waxy gene in corn
  • Development method and application of KASP molecular marker of waxy gene in corn

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Experimental program
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Effect test

Embodiment 1

[0028] Development of KASP marker closely linked to waxy related gene waxy in maize

[0029] 1. Primer design: According to the KASP_zmwx site, extract the flanking sequences of 100bp on the left and right sides, preferably use Primer5.0 software to design 3 sets of KASP primers, and select a set of KASP primers with good polymorphism after detection by the ArrayTape platform of Douglas Scientific Company For subsequent verification, the KASP primers used to detect KASP_zmwx are as follows:

[0030] Primer X: 5'-gaaggtcggagtcaacggattCTACAGGGACGCAAAGGT-3' (SEQ ID No.1, the lowercase part is the specific fluorescent tag sequence VIC);

[0031] Primer Y: 5'-gaaggtgaccaagttcatgctATCTACAGGGACGCCG-3' (SEQ ID No.2, the lowercase part is the specific fluorescent tag sequence FAM);

[0032] Primer R: 5'-TATACGAGCATGGAGAACGAA-3' (SEQ ID No.3);

[0033] 2. DNA extraction: Genomic DNA was extracted from corn leaves by conventional CTAB method;

[0034] 3. KASP reaction test

[0035] K...

Embodiment 2

[0054] Application of KASP marker closely linked to waxy related gene waxy in molecular marker-assisted selection of high waxy maize plants

[0055] In order to test the practicability of the KASP_zmwx marker of the present invention, the maize variety Chang 7-2 (as the female parent) described in Table 1 was crossed with Suyunuo 11 (as the male parent) to prepare an F2 segregation population, and the F2 segregation population was marked with KASP Detection and waxy phenotype record (test results are shown in Table 2), the implementation method of marker detection refers to Example 1; by statistics on the waxy phenotype of the test variety, in 48 isolated populations, only 2 individual plants The genotype does not match the waxy phenotype, and the consistency result between the waxy phenotype and the genotype is P=95.8% (P=the number of individual plants in the consistent population / the total number of the population); It has high practicability in the screening of sexual plan...

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Abstract

The invention discloses a development method and application of a KASP molecular marker of a waxy gene in corn, belonging to the technical field of biology. According to the invention, KASP specifically recognizes gene loci through a fluorescent probe to achieve a genotyping effect, and the KASP molecular marker can be used for detecting SNP loci and InDel loci. Compared with molecular markers such as SSR, RFLP and InDel, the KASP marker has the characteristics of rapidness in detection, low cost, easiness in large-scale application and the like; the KASP marker does not need to be typed according to the size of a DNA fragment, can get rid of the defects of tedious steps, low flux and relatively high price in a traditional gel electrophoresis detection method, and is more suitable for a high-flux molecular detection platform which rapidly develops at present. Therefore, development of the KASP molecular marker, which is low in cost and suitable for a high-throughput molecular detection platform, of the waxy gene in corn has important significance to promotion of application of a molecular marker technology and improvement of breeding efficiency and breeding level of waxy corn in China.

Description

technical field [0001] The invention relates to a development method and application of a KASP molecular marker of a corn waxy gene, and belongs to the field of biotechnology. Background technique [0002] Waxy maize is an important food crop in my country, and my country is one of the main birthplaces of waxy maize (Wang Yifa, Wang Liming, Shen Xuefang, etc. Origin, classification, variety improvement and industrial development of waxy maize [C]. Chinese Crop Society 2007 Proceedings of the National Symposium on Crop Genetics and Breeding. 2007.). The waxyness of corn is determined by the starch content, and the higher the amylopectin content, the higher the waxyness of corn. Because of its high amylopectin content, easy digestion and unique flavor, waxy corn is an important grain to improve people's living standards; at the same time, waxy corn is also an important feed plant, and raising milk cows with waxy corn can increase milk production; waxy corn is also an important...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 徐扬焦宇馨李成于广宁周恺杨文艳王欣杨泽峰徐辰武
Owner YANGZHOU UNIV