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Molecular marker for assistant breeding of bacterial blight resistant gene Xa21 and application of molecular marker

A bacterial blight resistance and molecular marker technology, which is applied in the fields of molecular biology and crop breeding, can solve the problems of tediousness, pollution electrophoresis detection, and the inability to apply large-scale breeding of transformed Xa21 materials, and achieves simple operation and improved breeding. Efficiency, short cycle effect

Active Publication Date: 2017-10-10
HUAZHI RICE BIO TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The transgenic policy has not been liberalized in my country, and transgenic Xa21 materials cannot be applied in actual large-scale breeding
The markers used in assisted selection are generally markers such as RAPD, SSR, AFLP, RFLP, CAP or dCAP, or based on the need for PCR or enzyme digestion or a combination of both, which require tedious electrophoretic detection with a risk of contamination

Method used

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  • Molecular marker for assistant breeding of bacterial blight resistant gene Xa21 and application of molecular marker
  • Molecular marker for assistant breeding of bacterial blight resistant gene Xa21 and application of molecular marker
  • Molecular marker for assistant breeding of bacterial blight resistant gene Xa21 and application of molecular marker

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Experimental program
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Effect test

Embodiment 1

[0026] Example 1 Obtainment of rice bacterial blight resistance gene Xa21 assisted breeding molecular markers

[0027] 1. Primer design

[0028]According to the relevant literature, the location of the Xa21 gene was determined in the 21273014-21277323 interval of rice chromosome 11, and the resequencing data of the Xa21 donor material and non-donor material (a total of 3000 rice materials) were used to determine the gene interval and its nearby sides Mining of SNP loci. The flanking sequences of the selected SNP sites were extracted, and primers were designed using the online primer design website BatchPrimer3 (http: / / probes.pw.usda.gov / batchprimer3 / ). Each group is marked with three primers, and the 5' ends of two specific primers are respectively connected with FAM and HEX fluorescent sequences. The primers were synthesized by Invitrogen Company.

[0029] The markers designed based on the KASP reaction principle and the single base difference of resistant and sensitive ma...

Embodiment 2

[0050] Example 2 Application of rice bacterial blight resistance gene Xa21 SNP marker RSXA211110

[0051] In order to test the specificity and practicability of the SNP marker RSXA211110 of the present invention, 188 materials were used to carry out natural population verification on the SNP marker RSXA211110. The 188 materials included varieties known to contain the homozygous Xa21 gene, other donors resistant to bacterial blight, general-sense materials, common hybrid rice and core rice breeding materials. Marked in natural group typing results such as figure 2 As shown, 7 cultivars known to contain the Xa21 gene were detected as homozygous Xa21 genotypes with resistance to bacterial blight, containing other gene donors resistant to bacterial blight, universal materials and Zhonghui 8015 (Xa21 donor ) core rice breeding materials were detected as homozygous xa21 genotype without bacterial blight resistance, and a few of the common hybrid rices were detected with heterozygo...

Embodiment 3

[0052] Example 3 Genetic mapping and phenotypic verification of SNP markers co-segregated with rice bacterial blight resistance gene Xa21

[0053] The F2 hybrid population of Xa21 donor parent Zhonghui 8015 and recipient parent Neixiang 5A was used to verify the genetic position and phenotype, which verified the feasibility and accuracy of the present invention again.

[0054] 1. Marker genetic position verification

[0055] Utilize the F2 population and 10 SNP markers with polymorphisms in the parents and the Xa21 specific marker RSXA211110 of the present invention to carry out genetic position verification, and the marker RSXA211110 of the present invention is mapped to the 21.2cM position of rice chromosome 11 ( image 3 ).

[0056] 2. Marker phenotype verification

[0057] The genetic phenotype of the marker RSXA211110 linked to the gene Xa21 of the present invention was verified by using Zhonghui 8015 and the F2 population of the recipient parent Neixiang 5A. At the ri...

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Abstract

The invention provides a molecular marker for assistant breeding of a bacterial blight resistant gene Xa21. The molecular marker is an SNP marker RSXA211110 co-separated with the rice bacterial blight resistant gene Xa21, and the SNP marker detects the 20894905-position base of chromosome 11 of rice; primers developed on the basis of aKASPtechnology comprise a specific primer X, a specific primer Y and a universal primer C, and sequences of the primers are shown as SEQ ID NO:1-3. The invention further provides an application of the SNP marker RSXA211110 to assistant breeding of the bacterial blight resistant gene Xa21. The application has the advantages of being simple to operate, low in cost and short in period, the marker is good in stability and is not affected by other gene effects and environmental factors, selection in early generation can be realized, the breeding period is shortened, the breeding efficiency is increased, and the molecular marker has great significance in improvement of bacterial blight resistant rice breeds and is suitable for assistant selective breeding of the gene Xa21.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and crop breeding, in particular to molecular markers for bacterial blight resistance gene Xa21 assisted breeding and applications thereof. Background technique [0002] Rice bacterial blight has physiological race specialization, and the resistance of varieties is basically controlled by the main resistance gene in the nuclear genome. The hybrid progenies were analyzed for their resistance to Japanese flora, and after the identification and naming of two dominant resistance genes, the identification and excavation of rice bacterial blight resistance genes have not stopped. After 1975, the International Rice Research Institute identified seven disease resistance genes using the Philippine race (Zhang Qi, 2007). Researchers from other countries have also identified a number of resistance genes one after another. However, due to the different bacterial strains used by scientists from var...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 彭佩郑秀婷江南梁毅贺治州李为国李继明肖金华
Owner HUAZHI RICE BIO TECH CO LTD
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