Method for directly identifying homologous recombination of higher plant DNA

A technology of homologous recombination and higher plants, applied in the field of plant genetics, can solve the problems of difficult identification of traits or a locus of homologous recombination, etc.

Active Publication Date: 2012-03-28
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a new method for directly identifying DNA homologous recombination in higher plants, aiming at the problem that existing plant homologous recombination detection methods are difficult to identify whether homologous recombination occurs for a pair of traits or a site. The method, which can determine whether homologous recombination occurs at this site and the recombination rate by detecting whether there is heterologous double-stranded DNA formation

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  • Method for directly identifying homologous recombination of higher plant DNA
  • Method for directly identifying homologous recombination of higher plant DNA
  • Method for directly identifying homologous recombination of higher plant DNA

Examples

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Embodiment 1

[0017] Example 1 Detection of homologous recombination of 'Zheyin No. 3 poplar' during sporocyte meiosis

[0018] 1. Test materials:

[0019] Maternal material to be tested: 'Zheyin 3 poplar' (P.pseudo-simonii×P.nigra'Zheyin3#');

[0020] Male material: 'Beijing Yang' (P.×beijingensis).

[0021] 2. Test methods and test results

[0022] When the embryo sac development stage of 'Zheyin No. 3 poplar' is in the uninucleate embryo sac stage, the embryo sac is subjected to physical and chemical treatment (soaking the female inflorescence 6-48 with 0.3-0.5% colchicine solution) hours) to obtain 2n female gametes with two sister chromosomes coexisting in the same gamete; cross the 2n female gametes with the male gametes of 'Beijing Poplar' (P.×beijingensis) to obtain triploid plants;

[0023] According to the poplar SSR database (http: / / www.ornl.gov / sci / ipgc / ssr_resource.htm), there are differences between 'Zheyin 3 poplar' and 'Beijing poplar', and 'Zheyin 3 poplar' Three pairs of...

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Abstract

The invention discloses a method for directly identifying homologous recombination of higher plant DNA, which comprises the following steps: inducing chromosome doubling of embryo sac by taking a plant to be identified as a female parent to obtain 2n female gametes; obtaining a hybrid triploid by the hybridization of the 2n female gametes and male gametes of a paternal plant; screening to obtain a codominant molecular marker primer; conducting PCR (Polymerase Chain Reaction) amplification with the screened codominant molecular marker primer; conducting capillary electrophoresis analysis to the amplification product: if only one of codominant molecular markers of the female parent occurs in the amplification product of the triploid, homologous recombination does not occur at relevant locusof the molecular marker, and if all genetic information of the female parent occur in the amplification product of the triploid, homologous recombination occurs to the pistillate parent at the relevant locus of the molecular marker and heteroduplex DNA is formed. The method has important theoretical and application values on plant genetic analysis, discrimination of molecule marker technical method and guidance on triploid breeding and the like.

Description

technical field [0001] The invention relates to a method for analyzing plant genetic traits, in particular to a method for directly identifying DNA homologous recombination in higher plants, belonging to the field of plant genetics. Background technique [0002] Meiosis is the turning point of the alternation of generations of sexual reproductive species, and the recombination between homologous chromosomes during meiosis diversifies the genetic basis of germ cells and accelerates the evolution of organisms. Although homologous recombination has been extensively studied, it remains one of the poorly understood phenomena of genetics. Since the 1960s, people have systematically studied the mechanism of homologous recombination in prokaryotes and fungi, and successively proposed a variety of homologous recombination models (Holliday R.A mechanism for gene conversion in fungi. Genet. Res, 1964, 5: 282-304; Meselson M S, Radding C M.A general model of genetic recombination. Proc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 康向阳董春波索玉静王君张平冬
Owner BEIJING FORESTRY UNIVERSITY
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