Molecular marker of rice-blast-resistant gene Pi2 and application of molecular marker

A rice blast resistance gene and molecular marker technology, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of difficult identification of long-lasting broad-spectrum disease-resistant varieties with multiple disease-resistant genes aggregated and low breeding efficiency , Inaccurate identification results and other problems, to achieve the effects of saving breeding screening costs, controlling the size of breeding groups, and improving accuracy

Inactive Publication Date: 2014-10-01
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Application Information

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Problems solved by technology

[0003] The breeding of traditional disease-resistant rice varieties mainly relies on the phenotypic identification of resistance inoculation at the seedling stage or the screening of resistance phenotypes in rice blast-prone areas, which is easily affected by changes in the environment, climate and other conditions of rice blast occurrence, resulting in

Method used

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  • Molecular marker of rice-blast-resistant gene Pi2 and application of molecular marker
  • Molecular marker of rice-blast-resistant gene Pi2 and application of molecular marker
  • Molecular marker of rice-blast-resistant gene Pi2 and application of molecular marker

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Experimental program
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Effect test

Embodiment 1

[0033] Analysis of the DQ352453 sequence and DQ454158 also found that in the DQ352453 sequence, PI2 Functional genetic terminal TGA downstream 2236 / 2237 BP has a sequence of 5'-GGCAGCGGGTCGGGGCGCGGGGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC, image 3 To.

[0034] Example 2 PI2 Establishment of functional genes upstream and downstream molecular marks

[0035] against PI2 Specific missing position polymorphism in the upstream interval of functional genes, designed a pair of specific primers PI2U-F: 5'-TATGGGGGGGTTGATTCC-3 ', PI2U-R: 5'-CTTCACAAGCCAACAaca-3' 3 ', for establishing upstream moleculesTag Pi2-Id-U.Use PI2U-F / PI2U-R primers to combine C101A51, Japan Qing, 9311, SE21S, Ming Row 63, Miyang 46, II-32B, Guangfu 998, 75-127, Test 64, 02428, Sanang B, B, B, B, B, B, B.The genome DNA templates of 17 rice varieties / products such as 63S, D62B, Shu Hui 527, spoke ...

Embodiment 2

[0036] The result of the electrophoresis shows that it contains PI2 The C101A51 amplification product of the functional gene shows a 144bp band, and the other 16 are PI2 Functional gene rice variety / product system amplification products (except 75-1-127 amplification products are 162 BP) display a 171bp or 172bp strip (attached figure 2 ), And the band type of the C101A51 sample and the belt type of other samples can be distinguished in 8%non -degeneration polyacrylamide gel.

[0037] against PI2 Specific missing position polymorphism in the downstream interval of functional genes, designed a pair of specific primers PI2D-F: 5'-Taggggcggggggac-3 ', PI2D-R: 5'-CTAGGCCCCCCCCCCCATAGT-3'Tag Pi2-Id-D.Use PI2D-F / PI2D-R primers to combine C101A51, Japanese Qing, 9311, SE21S, Mingjie 63, Miyang 46, II-32B, Guangfu 998, 75-127, Testing 64, 02428, Baogang B, B, B.The genome DNA templates of 17 rice varieties / products such as 63S, D62B, Shu Hui 527, spoke 838 and ZR101 are amplified.The PCR...

Embodiment 3

[0041] Example 4 PI2 Application of functional molecules marked in rice anti -disease molecular breeding

[0042] use PI2 Functional gene supply varieties C101A51 and Super Rice recovery system 9311 configuration combination, and through the combination of combined molecular marking auxiliary selection PI2 Functional genes to 9311.Obtained BC after the configuration, repayment and marking screening 3 Algebra.Further BC 3 Screening on behalf of the materials, extract BC 3 The genomic DNA of the agent, and the PCR analysis is detected separately PI2 The upstream label of the functional gene is PI2-Id-U and downstream label PI2-Id-D, and the samples of 24 plants are detected. PI2 The upstream PI2-ID-U band of the functional gene 144 BP is consistent with the results of the anti-disease biology identification of these plants.The results of the embodiment reflect the application of the present invention PI2 The molecular labeling method of functional genes can quickly and accurately id...

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Abstract

The invention belongs to the field of crop biotechnology and in particular relates to a molecular marker of a rice-blast-resistant gene Pi2 and application of the molecular marker. The upstream and downstream molecular markers are based on specific insertion/deletion loci polymorphism of nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2. The two molecular markers are co-dominant molecular markers developed for close linkage regions on two sides of the functional gene Pi2, and have the advantages of being high in accuracy and capable of being widely applied to different breeding parents. By detecting the two molecular markers, whether a detected rice material has the rice-blast-resistant gene Pi2 or not can be accurately judged. By virtue of a molecular marker combination, whether a hybrid transferred progeny plant genome of a parent of the functional gene Pi2 and other parents has the functional gene Pi2 or not and the homozygous state of the functional gene Pi2 can be accurately detected, and the breeding efficiency of a rice-blast-resistant rice variety with the functional gene Pi2 is improved.

Description

Technical field [0001] The present invention belongs to the field of crops and biotechnology, which involves rice resistance genes PI2 Molecular marks and applications. Background technique [0002] Rice is one of the most important grain crops in China and even the world, but its production is severely harmful to rice plague.In the world, the rice plastic bacteria ( Magnapotheezae ) The cause of rice plague can cause losses of 10-30%of rice output every year (Dean et al., 2005).Long -term production practice has shown that selection and use of disease -resistant rice varieties are the most effective, economical, and environmental safety methods to prevent and treat rice plague. [0003] The selection of traditional disease -resistant rice varieties mainly depends on the evaluation of the seedling inoculation phenotype of the seedling stage or the anti -episode form in the rice plastic disease is easy to be screened.The identification results are inaccurate and the breeding effic...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 陈松彪王锋田大刚陈在杰陈子强林艳
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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