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Method for identifying rice Wx-mw gene and application of rice Wx-mw gene in high-quality rice breeding

A wx-mw, rice technology, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of incomplete enzyme digestion, complicated operation of molecular markers, and difficulty in accurately identifying different genotypes.

Inactive Publication Date: 2015-11-25
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned molecular markers, especially the CAPS markers, are cumbersome to operate, and often have incomplete digestion, which makes it difficult to accurately identify different genotypes
The Wx-mw allele has not been reported so far, and there is no development of related molecular markers

Method used

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  • Method for identifying rice Wx-mw gene and application of rice Wx-mw gene in high-quality rice breeding
  • Method for identifying rice Wx-mw gene and application of rice Wx-mw gene in high-quality rice breeding
  • Method for identifying rice Wx-mw gene and application of rice Wx-mw gene in high-quality rice breeding

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Experimental program
Comparison scheme
Effect test

Embodiment 1W

[0038] The cloning of embodiment 1Wx-mw gene

[0039] In our previous research, we found that the amylose content of an indica rice germplasm Mowanggu (provided by Hunan Hybrid Rice Research Center) was significantly lower than that of indica rice Nanjing 11 (Jiangsu Province mid-term bank of agricultural germplasm resources) and between the known conventional japonica rice 2661 (selected strain of the Agricultural College of Yangzhou University) and dark endosperm rice Nanjing 46 (medium-term bank of agricultural germplasm resources of Jiangsu Province) ( figure 1 ). Since the amylose content is mainly controlled by the Wx gene, the gene was amplified by PCR and sequenced using high-fidelity DNA polymerase using the genomic DNA of Mowanggu as a template. The 8073bp genome sequence of the gene was analyzed. Through sequence comparison, the Wx gene derived from the Devil's Valley and the known alleles wx, Wx-a, Wx-b, Wx-in, Wx-mq, Wx-mp and Wx -op(Wx-hp) sequences are all con...

Embodiment 2

[0041] Example 2 Design and detection of molecular markers of four-primer amplification hindered mutation system

[0042] According to Example 1, compared with the Wx-a and Wx-in genes, the Wx-mw gene has a G-T single nucleotide variation site at the first intron splice site, specifically in the Wx-mw allele The second base of the first intron is T and in the Wx-a and Wx-in alleles it is G. According to this difference, specific four primers were designed to amplify the molecular markers of the hindered mutant system (external primers Wx-mw-out1-FSEQIDNO.1 and Wx-mw-out1-RSEQIDNO.2 and internal primers Wx-mw-in1-FSEQIDNO. 3 and Wx-mw-in1-RSEQIDNO.4) were used to verify this mutation site ( image 3 ). For distinction, this molecular marker is called molecular marker 1.

[0043] According to Example 1, compared with wx, Wx-a, Wx-b, Wx-mq, Wx-mp and Wx-op alleles, there is a special single nucleotide variation site in the sixth exon, specifically It is C in the sixth exon 62...

Embodiment 3

[0049] Example 3 Molecular marker-assisted breeding of new rice lines with low amylose content

[0050]The present invention provides an idea to carry out continuous backcross breeding with rice plants carrying Wx-mw genes (such as rice devil valley) as donor parents and rice varieties or lines to be improved (such as 2661) as recurrent parents. In the low generation, the molecular marker 2 linked with the target gene Wx-mw gene and its control low amylose content traits in Example 2 is used for assisted selection. In the high generation population, it can be selected in combination with the amylose content. After a certain number of generations, a new variety or line can be bred that not only retains the excellent traits of the original parent to be improved, but also has excellent eating quality with low amylose content.

[0051] The invention introduces the Wx-mw gene into the japonica rice line 2661. Specifically, Mowanggu (an indica rice variety carrying the Wx-mw gene) ...

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Abstract

The invention relates to a method for identifying a rice Wx-mw gene and application of the rice Wx-mw gene in high-quality rice breeding. The invention discloses application of the second base of the first intron of a Wx-mw allele and the base of the sixth exon 62bp of the Wx-mw allele in identifying the rice Wx-mw gene as molecular markers. Four labeled primers are respectively designed according to splice site mutation of the first intron and a mutated site on the sixth exon, and the designed four labeled primers are respectively added to a same PCR (polymerase chain reaction) system so as to amplify various rice DNAs. To distinguish Wx-mw from Wx-a and Wx-in, the Wx-mw is introduced into a target variety through auxiliary selection by virtue of the two four-primer molecular markers and in coordination with conventional crossing and back-crossing; and the method is suitable for breeding a novel rice variety which is relatively low in amylose content and excellent in eating quality.

Description

technical field [0001] The invention belongs to the technical field of plant breeding, and specifically relates to a four-primer PCR molecular marker method for identifying different genotypes of rice low-amylose content mutant gene Wx-mw and a method for cultivating rice varieties with excellent eating quality by using the molecular marker. Background technique [0002] As an important food crop in my country and even in the world, rice plays an important role in ensuring food security in my country. In recent decades, my country's rice production has continued to increase, especially with the improvement of national consumption levels and living tastes, there is an increasing demand for diversified high-quality rice. [0003] The quality of rice is diverse. As far as high-quality food is concerned, it mainly includes four aspects: milling quality, appearance quality, cooking and eating quality, and nutritional quality. These quality traits directly determine the commodity ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 刘巧泉张昌泉李钱峰周兴忠顾铭洪
Owner YANGZHOU UNIV
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