Anti-rice blast gene Pi9 specific CAPS marker Pi9caps and application thereof

A rice blast resistance gene and rice blast technology, applied in the field of crop molecular genetics and breeding, can solve problems such as low selection efficiency, difficulty in rice blast resistance genes, and high chance of polymorphism

Inactive Publication Date: 2015-05-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, since many restriction endonucleases can participate in the digestion of amplified DNA, the chance of detecting polymorphisms is greater
[0004] It is very difficult to use the conventional field phenotype to identify the selection of the rice blast resistance gene, the selection efficiency is very low, and the selection accuracy is very poor. However, the use of molecular markers linked to the rice blast resistance gene or the specific marker selection of the gene itself is very difficult in production. Proven to be very effective and economical
For the selection of molecular markers of the Pi9 gene, researchers have developed some markers, but they all have some problems or inconvenie

Method used

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  • Anti-rice blast gene Pi9 specific CAPS marker Pi9caps and application thereof
  • Anti-rice blast gene Pi9 specific CAPS marker Pi9caps and application thereof
  • Anti-rice blast gene Pi9 specific CAPS marker Pi9caps and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073]Example 1, Primer Design and Detection of Rice Blast Resistance Gene Pi9 Gene-Specific CAPS Marker Pi9caps

[0074] 1. Analysis of Pi9 gene sequence-specific SNP sites

[0075] The published Pi9 gene and the sequenced genome sequences of the corresponding regions of rice varieties 93-11 and Nipponbare were downloaded from the public database, and the sequences were compared against the Pi9 locus to screen for Pi9-specific genes that could be differentiated from those genes. SNPs for locus alleles. Wherein the T base located at 7080bp after the start codon of the Pi9 gene (that is, the nucleotide sequence of 75-1-127BAC12.1 in rice chromosome 6 from the 5' end to the 40804th position) is at 93-11 and Nipponbare The alleles in are all C, and this SNP is just located in the recognition sequence of restriction endonuclease XbaI (recognition sequence is 5'-TCTAGA-3') (such as figure 1 as shown, figure 1 Among them, Pi9 is the rice blast resistance gene Pi9, Pi9-9311 is the...

Embodiment 2

[0097] Example 2, Verification and Application of Rice Blast Resistance Gene Pi9-specific CAPS Marker

[0098] 1. Using the rice variety 93-11 without the blast resistance gene Pi9 as the female parent, and the rice variety 75-1-127 containing the blast resistance gene Pi9 as the male parent, the F1 generation was obtained, and the F1 generation was obtained by selfing For the F2 population, extract the genomic DNA of individual plants numbered 4-24, 93-11 and 75-1-127 in the F2 population.

[0099] Two, the genomic DNA obtained in step 1 is used as a template respectively, and the primers F and R of Example 1 are used as primers to carry out PCR amplification to obtain a PCR amplification product with a size of 802bp. This PCR amplification product is the Pi9caps marker , the marker is the 75-1-127BAC12.1 sequence from the 5' end to the 41006th position of the 75-1-127BAC12.1 sequence in the No. 6 chromosome of each rice, and then use the restriction endonuclease XbaI to dige...

Embodiment 3

[0106] Example 3. Verification and application of rice blast resistance gene Pi9-specific CAPS marker in rice transfected with Pi9 gene coding region

[0107] 1. Using the rice variety TP309 without the rice blast resistance gene Pi9 as a transgenic acceptor, overexpressing the CDS region of the rice blast resistance gene Pi9 in TP309 to prepare rice transgenic for the coding region of the Pi9 gene.

[0108] Two, the genomic DNA of the rice and TP309 of the transgenic Pi9 gene coding region that extraction step 1 obtains, use it as template respectively, take the primer F and R of embodiment 1 as primer, carry out PCR amplification, obtain each PCR amplification product, Restriction endonuclease XbaI was used to digest each PCR amplification product to obtain each digested product, and each digested product was detected by electrophoresis on an agarose gel.

[0109] The result is as Figure 4 shown.

[0110] Figure 4 Among them, lane 1 is the DNA ladder, and lanes 2 and 3 ...

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Abstract

The invention discloses an anti-rice blast gene Pi9 specific CAPS marker Pi9caps and application thereof. The invention discloses a method for identifying or auxiliarily identifying whether rice contains anti-rice blast gene Pi9 and/or whether rice has rice blast resistance, which comprises the following steps: detecting 40804th nucleotide from the 5' terminal of 75-1-127BAC12.1 sequence or homologous sequence thereof in chromosome 6 of rice to be detected, and identifying according to the kind of the 40804th nucleotide the 5' terminal of the 75-1-127BAC12.1 sequence or homologous sequence thereof in chromosome 6 of rice. The anti-rice blast gene Pi9 specific CAPS marker Pi9caps can accurately judge whether the selected rice plant contains the anti-rice blast gene Pi9, and thus, has important application meanings in accelerating the anti-rice blast rice species breeding progress.

Description

technical field [0001] The invention relates to a specific CAPS marker Pi9caps of the rice blast resistance gene Pi9 and an application thereof, belonging to the field of crop molecular genetics and breeding. Background technique [0002] Rice is one of the most important food crops in the world, and more than half of the world's population uses rice as a staple food. Rice blast is the main disease that affects rice production, ranking first among fungal diseases. Rice blast causes a 10% to 30% reduction in rice production worldwide every year, and in severe cases, a 40% to 50% reduction in production, or even no harvest. Practice has proved that the use of rice blast resistance genes is the most economical, effective and environmentally friendly method to control rice blast disease. So far, at least 68 rice blast resistance loci and 83 major genes have been reported in rice. These genes are distributed in clusters on all rice chromosomes except chromosome 3, and 24 have be...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 朱立煌吕启明黄志远盘毅唐丽李晓兵周壮志辛业芸赵炳然符习勤
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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