Method for identifying psathyrostachys huashanica allogenic material in wheat genome

An identification method and technology for exogenous substances are applied in the field of identification of exogenous substances in Huashan new wheatgrass, which can solve the problems of cumbersome enzyme extraction procedures, different contents in developmental stages, difficult operation and the like, and achieves simple operation, high specificity and repeatability. strong effect

Inactive Publication Date: 2013-08-14
NORTHWEST A & F UNIV
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Problems solved by technology

However, this technique is not easy to operate, and the enzyme extraction procedure is cumbersome, and the...

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  • Method for identifying psathyrostachys huashanica allogenic material in wheat genome
  • Method for identifying psathyrostachys huashanica allogenic material in wheat genome
  • Method for identifying psathyrostachys huashanica allogenic material in wheat genome

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Embodiment Construction

[0028] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0029] The genetic resource of the present invention, Psathyros tachys huashanica Keng2n=2x=14, NsNs, is collected in Shaanxi Province, China.

[0030] A method for identifying exogenous substances of P. chinensis in wheat genome, comprising the following steps:

[0031] 1 Screening of RAPD-specific sequences and recovery of specific fragments of P.

[0032] Using the genomic DNA of P. huashanensis and various wheat relatives as templates, 500 RAPD10-base random primers were used for PCR amplification, and the polymorphic bands of P. huashanensis were screened and recovered.

[0033] 20 μL total PCR reaction system includes: 10×PCR Buffer 2 μL, dNTPs 2.5 μmol / ml 2 μL, primer 2.5 μmol / ml 1 μL, Mg 2+ 2.5mmol / ml 1μL, template DNA 40-60ng / μL 2μL, Taq enzyme 5U / μL 0.2μL, ddH 2 O11.8 μL.

[0034] Reaction program: pre-d...

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Abstract

The invention discloses a method for identifying a psathyrostachys huashanica allogenic material in a wheat genome. The method comprises the following steps of: carrying out psathyrostachys huashanica specific sequence screening by an RAPD (random amplified polymorphic deoxyribonucleic acid) primer; recovering, cloning, sequencing and comparing specific strips; carrying out Southern analysis, converting a psathyrostachys husshanica SCAR (sequence characterized amplified region) mark by primer design; carrying out specific detection on the SCAR mark by utilizing the near-source wheat species genome; and finally identifying the entire chromosome of the psathyrostachys huashanica in a wheat-psathyrostachys huashanica complete alien addition line and an alien addition line with undetermined homologous group relation. By adopting the method, genetic resources of the psathyrostachys husshanica are fully utilized; the identification method of the psathyrostachys husshanica allogenic material in the wheat genome is simplified; the utilization efficiency of an exogenous genetic resource is improved; and reliable guarantee is also provided. The method disclosed by the invention is simple and feasible, and suitable for popularization and application.

Description

technical field [0001] The invention belongs to the field of agricultural technology, and relates to a method for identifying exogenous substances of P. chinensis in wheat genome. Background technique [0002] In situ hybridization (in situ Hybridizztion, ISH) technology is the product of the successful combination of cytology and modern molecular biology. It is currently the most widely used in the identification of foreign substances. The exogenous nucleic acid probe labeled with radioactive substances hybridizes with the specific chromosome on the chromosome preparation, amplifies the hybridization signal, and detects it by autoradiography or other detection systems to determine the heterologous chromosome or its fragment of a certain species . Specific methods include: 1 Chromosome preparation, 2 Probe and blocking DNA preparation, 3 DNA detection, 4 DNA shearing, 5 Digoxigenin labeling DNA program by nick translation, 6 Denaturation of chromosome preparation, 7 Hybridi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 武军杜万里陈新宏赵继新杨群慧刘淑会王婧庞玉辉
Owner NORTHWEST A & F UNIV
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