Molecular marking method of rice stigma exsertion major QTL sites

A technology of exposed stigma and molecular markers, applied in the field of molecular genetics and breeding, can solve the problems of retention, fine positioning and cloning of non-main QTLs, etc., and achieve the effects of controlling the breeding scale, quick and easy identification methods, and high selection efficiency

Inactive Publication Date: 2014-12-03
国家粳稻工程技术研究中心 +1
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Problems solved by technology

The above studies are only at the preliminary exploration stage of the mapping of rice stigma exposed QTL, but have not carried out fine mapping and cloning of a major QTL

Method used

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  • Molecular marking method of rice stigma exsertion major QTL sites
  • Molecular marking method of rice stigma exsertion major QTL sites
  • Molecular marking method of rice stigma exsertion major QTL sites

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Experimental program
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Embodiment Construction

[0021] (1) Materials and methods:

[0022] 1. Materials: High stigma exposed germplasm DS was crossed with japonica rice variety C9083, the hybrid was planted, and F2 segregation population was obtained by selfing, which was used for the detection and positioning of stigma exposed QTL.

[0023] 2. Extract individual genomic DNA by SDS method.

[0024] 3. Screening of polymorphic markers: 600 pairs of SSR primers were used to perform PCR amplification with DS and C9083 DNA as templates to screen for polymorphic markers.

[0025] 4.PCR reaction: the volume is 10μl, in which 10×PCR Buffer (25mmol / LMg 2+ ) 1 μl, primer pair (2 μM) 1 μl, dNTP (10 mM) 0.2 μl, Taq enzyme (5U / μl) 0.25 μl, template (10-100ng / μl) 1.5 μl, add water to 10 μl. The PCR reaction program was pre-denaturation at 94°C for 2 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 30 s, and 35 cycles; finally, extension at 72°C for 10 min. The amplified products were separated by ...

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Abstract

The invention belongs to the field of molecular genetics and breeding, and discloses a molecular marking method of rice stigma exsertion major QTL sites. Development of a new molecular marker and fine mapping of rice stigma exsertion rate QTL are carried out for a target zone of third chromosome, PCR amplification of rice genome DNA is carried out by using one or two pairs of primers of Indel marked primer GS09 and SSR marked primer RM15206, and the above obtained PCR amplification products are detected through electrophoresis on 8% native polyacrylamide gel. A qPES-3b synergistic site exists if a corresponding size of a fragment is amplified. The rice high stigma exsertion germplasm and breeding populations constructed by all parents can be detected through tightly linked molecular markers controlling the stigma exsertion major QTL sites, so the major QLT controlling the stigma exsertion can be rapidly introduced, the selection efficiency of major sites is improved, and the improvement of hybrid rice sterile line stigma exsertion and the improvement of the seed yield are benefited.

Description

technical field [0001] The invention belongs to the field of molecular genetics and breeding, and relates to a molecular marker method for rice stigma exposure main effect QTL sites, which is mainly used for molecular marker-assisted improvement of stigma exposure height of hybrid rice sterile lines and innovative utilization of germplasm resources. Background technique [0002] The development of hybrid japonica rice in northern my country has been slow because of poor outcrossing and low seed production yield. The exposed stigma rate of the male sterile line is the key factor determining the yield of hybrid seed production in the three-line and two-line breeding systems, and the exposed stigma of rice has always been an important trait in the breeding of the male sterile line. Trying to increase the stigma exposure rate of the Japonica CMS line during the breeding process will effectively improve the outcrossing characteristics of the Japonica CMS line, increase the yield ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q1/6858C12Q2600/156
Inventor 刘欣亓娜米佳佳王汝华李志彬华泽田
Owner 国家粳稻工程技术研究中心
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