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Method for identifying germplasm resource of radix codonopsis by use of ISSR (inter-simple sequence repeat) fingerprint

A technology of fingerprints and Codonopsis pilosula, which is applied to biochemical equipment and methods, and the determination/inspection of microorganisms, etc., can solve the problems of complex operation steps, high cost of identification methods, and long time consumption, and achieve the optimization of amplification procedures, simple and easy Effect of implementing and reducing workload

Inactive Publication Date: 2018-05-01
湖北省农业科学院中药材研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to solve the technical problems of the existing identification methods of medicinal plants, such as high cost, complicated operation steps, and long time-consuming

Method used

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  • Method for identifying germplasm resource of radix codonopsis by use of ISSR (inter-simple sequence repeat) fingerprint
  • Method for identifying germplasm resource of radix codonopsis by use of ISSR (inter-simple sequence repeat) fingerprint
  • Method for identifying germplasm resource of radix codonopsis by use of ISSR (inter-simple sequence repeat) fingerprint

Examples

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Embodiment 1

[0023] This embodiment discloses a method for identifying germplasm resources of Codonopsis pilosula using ISSR fingerprints. Those skilled in the research field can refer to the description of this embodiment, or perform technical means (such as test parameters, etc.) replacement or appropriate changes and combinations based on the method described in this embodiment to realize the present invention in a more preferred manner.

[0024] The method for identifying Codonopsis pilosula germplasm resources using ISSR fingerprints provided in this example includes such operational steps as acquisition of Codonopsis pilosula plant resources, DNA preparation, ISSR-PCR amplification, and agarose gel electrophoresis detection. Regarding the acquisition of Codonopsis pilosula plant resources, two methods are usually adopted, one is to transplant the whole Codonopsis pilosula from different origins; the other is to collect the seeds of Codonopsis pilosula plants from different origins for...

Embodiment 2

[0050] On the basis of Example 1, this example provides an identification method for Codonopsis pilosula samples to be tested. The sample to be tested was taken to construct an ISSR fingerprint according to the method described in Example 1, and the ISSR fingerprint of the sample to be tested was compared with the ISSR fingerprint of the above-mentioned 7 origins of Codonopsis pilosula. The ISSR fingerprints of the samples to be tested are shown in Table 6. It can be seen from Table 6 that sample 1 is consistent with the ISSR fingerprint of Codonopsis pilosula in Hadapu area, Gansu Province, and identified as the Codonopsis pilosula species in Hadapu area; while sample 2 and the ISSR fingerprint of Codonopsis pilosula from 7 production areas constructed by primer UBC844 After comparing the spectra, no matching fingerprints were found, which proves that the Codonopsis pilosula does not belong to the 7 production areas.

[0051] Table 6 ISSR (primer UBC844) fingerprints of Codo...

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Abstract

The invention discloses a method for identifying the germplasm resource of radix codonopsis by use of ISSR (inter-simple sequence repeat) fingerprint. The method comprises the following steps: (1) collecting and numbering radix codonopsis from different production places; extracting the DNA of the radix codonopsis from different production places and performing PCR amplification; detecting amplification products by agarose gel electrophoresis; according to the result of agarose gel electrophoresis and by adopting the number of radix codonopsis as a horizontal coordinate and the length of DNA fragment as a longitudinal coordinate, recording the detection results of the DNA amplification stripes of the radix codonopsis from different production places at the intersections between the horizontal coordinate and the longitudinal coordinate respectively; marking the detection result with DNA amplification stripe, and not marking the detection result without DNA amplification stripe, so as toobtain the ISSR fingerprints of the radix codonopsis from different production places; (2) comparing the ISSR fingerprint of the to-be-detected radix codonopsis with the ISSR fingerprints of the radix codonopsis from different production places, and judging the production place of the to-be-detected radix codonopsis according to the comparison result. The method disclosed by the invention can beused for conveniently and quickly identifying the radix codonopsis medicinal materials with little difference in appearance and easily confused, and quick and accurate identification can be performedin the seedling stage of the radix codonopsis; therefore, the method is of great significance on guaranteeing the accuracy and reliability of the base source of a medicinal material.

Description

technical field [0001] The invention belongs to the technical field of quality detection of traditional Chinese medicinal materials, and relates to a method for identifying origins of Codonopsis pilosula, in particular to a method for identifying germplasm resources of Codonopsis pilosula using ISSR fingerprints. Background technique [0002] Codonopsis pilosula is the dry root of Codonopsis pilosula (Franch.) Nannf), C.pilosula Nannf.var.modesta (Nannf.) L.T.Shen or C.tangshenOliv. The commonly used traditional Chinese medicine for medicine and food in my country has the effects of invigorating the spleen and lungs, nourishing blood and promoting body fluid. Driven by economic interests and limited by the level of species identification, Codonopsis pilosula is currently used in many places, and the value of different varieties is different. The aboveground parts of Codonopsis pilosula belong to closely related plants in appearance, and the types of active ingredients contai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2531/113C12Q2565/125
Inventor 卢超张美德何银生艾伦强刘海华
Owner 湖北省农业科学院中药材研究所
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