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Function-specific molecular markers of rice high-yield allele gy6

An allele and molecular marker technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of false positives, failure to achieve the expected goal of improvement, deletion of target genes, etc., and saves money Time and cost, improved breeding efficiency, effect of increased screening accuracy

Active Publication Date: 2016-08-17
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of functional markers for MAS can overcome some disadvantages brought about by linked molecular markers: 1) Genetic burden, that is, the target gene (favorable gene) is linked with its nearby non-target gene (unfavorable gene), and the target gene is also introduced. Unfavorable genes were brought in, resulting in improvement that failed to achieve the expected goal; 2) The marker information was misleading, that is, the marker still existed, but the marker was separated from the gene due to recombination, resulting in the loss of the target gene, resulting in false positives

Method used

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  • Function-specific molecular markers of rice high-yield allele gy6
  • Function-specific molecular markers of rice high-yield allele gy6
  • Function-specific molecular markers of rice high-yield allele gy6

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Development of specific molecular markers for GY6 gene

[0020] 1. GY6 allele amino acid sequence alignment and identification of specific amino acid positions

[0021] The sequence alignment software DNASTAR MegAlign module (Lasergene) was used to analyze the amino acid sequence of Zhenshan 97 and Miyang 46 at the GY6 locus (Patent No.: ZL 201210047301.0, SEQ ID No: 3 and SEQ ID No: 6) . The GY6 protein was identified as having differences in 6 amino acid positions, namely A31V, K105E, N144S, K146R, D147N and A160T ( figure 1 ).

[0022] 2. Design of GY6 gene-specific molecular marker primers

[0023] Aiming at the 31st amino acid position among the above 6 amino acid differences, according to the CAPS marker design principle, the online software dCAPS Finder 2.0 (http: / / helix.wustl.edu / dcaps / dcaps.html) and Oligo7.0 are used. (Molecular Biology Insights) Design primers. The upstream primer sequence is 5'-CCGGATCACTAACCTCAGCG-3', and the downstream primer sequence...

Embodiment 2

[0041] Example 2 Verification of GY6 gene-specific molecular markers in near-isogenic populations

[0042] Taking a set of near-isogenic populations derived from Zhenshan 97 / Miyang 46 as materials, this population segregated only in the GY6 interval, containing two genotypes of Zhenshan 97 and Miyang 46. Compared with the latter, the former is true It has a large number of grains, a thousand grains, and a high yield. The GY6 gene specific molecular marker Si2926 / Hha I was tested for function. Using the PCR amplification system and reaction conditions described in Example 1, Si2926 / Hha I was used for amplification and restriction digestion.

[0043] Electrophoresis results ( image 3 ) Showed that Si2926 / Hha I was segregated in the near-isogenic line population. The strains (1-15) with a large number of real grains, a thousand grains and a high yield were all detected with the same band pattern as Zhenshan 97, indicating that they All of them carried the Zhenshan type 97 high-yield...

Embodiment 3

[0044] Example 3 Detection of GY6 gene-specific molecular markers in rice germplasm resources

[0045] Using the functional specific molecular marker Si2926 / Hha I of GY6 gene, 66 indica hybrid rice parents were tested. The results showed that 17 parents carried Zhenshan type 97 alleles and 49 parents carried Miyang type 46 alleles ( Figure 4 ).

[0046] China National Rice Research Institute

[0047] Function-specific molecular markers of rice high-yielding allele GY6

[0048] 2

[0049] PatentIn version 3.3

[0050] 1

[0051] 20

[0052] DNA

[0053] Artificial sequence

[0054] 1

[0055] ccggatcact aacctcagcg 20

[0056] 2

[0057] twenty one

[0058] DNA

[0059] Artificial sequence

[0060] 2

[0061] gcatccaaga tatgatccgt a 21

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Abstract

The present invention provides a function-specific molecular marker Si2926 / HhaI for detecting rice high-yielding allele GY6, its upstream primer sequence is shown in SEQ ID No: 1, and its downstream primer sequence is shown in SEQ ID NO.2. Using this The PCR amplification and restriction endonuclease digestion of the DNA extracted from the rice seedling stage by the primers can detect whether the material to be tested carries the rice high-yield allele GY6, and the whole detection process is simple and accurate.

Description

Technical field [0001] The invention relates to the technical field of detection in rice breeding, in particular to a functional specific molecular marker of the high-yield allele GY6. Background technique [0002] Rice is one of the most important food crops in the world. Increasing the yield of rice is of great significance to solving the problem of food security. Rice yield per plant is determined by three constituent factors: the number of ears per plant, the number of grains per ear, and the thousand-grain weight, which are all complex quantitative traits and are controlled by multiple quantitative trait loci (QTL). In recent years, with the completion of rice genome sequencing, the cloning of QTLs for rice yield traits has made rapid progress. 16 control plant types, panicle shapes, number of grains per panicle, number of spikelets per panicle, and thousand-grain weight have been cloned. QTLs for traits related to rice yield. However, relative to the complexity of yield ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 樊叶杨庄杰云朱玉君
Owner CHINA NAT RICE RES INST
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