Molecule identification method for Qingdao zoysia japonica and zoysia matrella filial generation authenticity

A technology of Zoysia sativa and identification method, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of long time, poor accuracy, and high consumption, and achieve accuracy High, evenly distributed, highly polymorphic effects

Inactive Publication Date: 2008-08-13
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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Problems solved by technology

[0005] Technical problem The purpose of the present invention is to solve the field identification detection method and other molecular identification of the authenticity of Zoysia plant hybrid progeny in the prior art, which takes a lo

Method used

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  • Molecule identification method for Qingdao zoysia japonica and zoysia matrella filial generation authenticity
  • Molecule identification method for Qingdao zoysia japonica and zoysia matrella filial generation authenticity
  • Molecule identification method for Qingdao zoysia japonica and zoysia matrella filial generation authenticity

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Experimental program
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Embodiment Construction

[0036] (1) Screening of primers

[0037] (1) Preparation of materials: Zoysia japonica Z022 (Z.japonica Steud.cv.Qingdao) and Zoysia japonica Z123 (Z.matrella) (see reference Liu Jianxiu et al. Preliminary research on [J]. Acta Prataphysica Sinica, 2003, 12(2): 70-75; Kenneth B.Marcum, Sharon J.Anderson, M.C.Engelke.SaltGland Ion Secretion: A Salinity Tolerance Mechanism among Five Zoysiagrass Species[J] .Crop Sci, 1998, 38: 806-810) 9 hybrid progeny seeds were treated with 30% NaOH for 80 minutes, cultivated and germinated in a petri dish, and when the seedlings grew to 1.5-2cm, they were moved into the hole tray, and there was a certain Transplant it to the flower pot when the biomass is full, and transplant it to the test site when the plate is full.

[0038] (2) Extraction of genomic DNA from materials: SDS method (Sodium Dodecyl Sulfate, sodium dodecyl sulfate) was used. Grind the leaves (generally about 3 grams) in liquid nitrogen, transfer to a frozen centrifuge tube,...

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Abstract

The invention relates to a method for identifying authenticity of filial generation of zoysia and manilagrass, which belongs to the field of biological technology. The invention is specially for rapid and precise identification of authenticity of filial generation of Qingdao zoysia and manilagrass. Using a genome DNA template of nine zoysia filial generations and two parents Qingdao zoysia and manilagrass as the template, select three pairs of SRAP primers, namely primers capable of amplifying specific band not included in female parent on male parent. Perform amplifying and electrophoresis on the male parent, the female parent and the filial generation, and identify according as whether the F1 generation can amplify male parent specific band. The result shows that a filial generation is a real hybrid when each of the nine filial generations has male parent specific band. Thus, a molecular identification system of authenticity of filial generation of Qingdao zoysia and manilagrass is established, by which the authenticity of filial generation of Qingdao zoysia and manilagrass can be quickly identified, the method can also be used for hybrid variety fingerprint map construction and purity identification.

Description

1. Technical field [0001] The invention relates to a molecular identification method for the authenticity of hybrid progeny of plants belonging to the genus Zoysia, belonging to the field of biotechnology. It is specially used for rapid and accurate detection of the authenticity identification of the hybrid offspring of Zoysia qingdao and Zoysia ditchleaf. 2. Background technology [0002] As a traditional breeding method, cross-breeding is quite mature, and it is a commonly used technical strategy in plant genetic improvement. Cross-breeding using parents with obvious differences in turfgrass traits is an important way to breed high-quality and excellent new strains. The flowering habit of Zoysia chinensis is that the pistils ripen first and the stamens ripen later. The research of Guo Hailin et al. shows that the self-seed setting rate of Zoysia grass is 0.0-54%, with an average of 16.15%, so the hybrid offspring may be mixed with some self-bred species . Accordingly, it...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 刘建秀薛丹丹郭海林郭爱桂
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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