Construction and identification method of molecular marking fingerprint of Dendrobium huoshanense and similitude species thereof

A Dendrobium Huoshanense, fingerprinting technology, applied in the construction and identification of Dendrobium germplasm fingerprints, can solve the problems of high cost, long time-consuming, complicated procedures, etc., and achieve simple steps, optimized reaction system, and strong genotyping ability Effect

Inactive Publication Date: 2009-06-10
陈乃富
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of high cost, complicated procedures and long time-consuming methods for identifying medicinal plants in the prior art, the present invention provides a molecular marker fingerprint of Dendrobium Huoshanense and its similar species that is easy and quick to identify, has good repeatability and high resolution. Atlas construction and identification methods

Method used

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  • Construction and identification method of molecular marking fingerprint of Dendrobium huoshanense and similitude species thereof
  • Construction and identification method of molecular marking fingerprint of Dendrobium huoshanense and similitude species thereof
  • Construction and identification method of molecular marking fingerprint of Dendrobium huoshanense and similitude species thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0037] In this example, the collected leaves of 8 Dendrobium strains that are similar to Dendrobium Huoshanense in terms of medicinal value and morphology are used as test materials, the total DNA of the genome is extracted, and the ISSR molecular marker analysis of the specified primers is carried out. The presented bands established ISSR-PCR molecular marker fingerprints of each test sample, and the constructed ISSR-PCR molecular marker fingerprints were used for germplasm identification of Dendrobium.

[0038] The specific operation steps are as follows:

[0039] 1), collecting fresh dendrobium sample pretreatment;

[0040] Consume starch with fresh Dendrobium dendrobium more than 6 hours;

[0041] Take 10 grams of fresh dendrobium leaves after dark treatment, put more than 50 grams of acetone and water mixed solution with a ratio of 3:1 and store for two days, take out the materials, soak in more than 50 grams of distilled water for one hour, and then take out the materia...

Embodiment 2

[0082] Identification of Dendrobium Huoshanenseensis by the DNA Fingerprints of Eight Dendrobium Lines Constructed

[0083] Get the unknown sample to establish fingerprints according to the method in Example 1, compare the fingerprints of the unknown samples with the obtained DNA fingerprints of the 8 strains of Dendrobium, and if it is the same as the Dendrobium Huoshanense spectrum, it is Dendrobium Huoshanense, otherwise it is not Dendrobium Huoshanense.

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Abstract

The invention relates to a method for constructing and identifying molecular marker fingerprint chromatogram of Dendrobium huoshanense and similar species thereof, which solves the problems that a method of identifying medicinal plants in the prior art has high cost, complicated procedures and long time consumption. The method comprises the steps of: 1, the collection of Dendrobium huoshanense samples; 2 the extraction and purification of DNA of genomes of the samples; 3. ISSR-PCR amplification; 4, agarose gel electrophoresis; 4, the construction of ISSR molecular marker fingerprint chromatogram of the samples to be tested; and 5, the identification of Dendrobium huoshanense germplasm by utilizing the constructed ISSR molecular marker fingerprint chromatogram. The method has the advantages that the method saves the time and the cost, and can obtain an accurate and reliable identification chromatogram through the extraction to DNA of Dendrobium plants, the ISSR-PCR amplification, and the agarose gel electrophoresis; the method has the advantages of simplicity, convenience, quickness, good repetitiveness, and high resolution on the identification of raw materials which are easy to confuse in appearance; and the method can identify in a seedling stage, which has important effect on ensuring the accuracy and stability of base resources of medicinal materials.

Description

technical field [0001] The invention relates to the construction and identification method of dendrobium germplasm fingerprints. Background technique [0002] There are three kinds of Dendrobium produced in Huoshan that can be processed into high-quality maple buckets, and Huoshan Dendrobium is the most popular. Dendrobium huoshanense C.Z.Tang et S.J.Cheng, also known as Dendrobium huoshanense C.Z.Tang et S.J.Cheng, is a kind of medicinal Dendrobium mainly containing mucopolysaccharides. Dendrobium candidum (Dendrobium candidumWall.ex Lindl.) from Huoshan is the most famous herb in China. Dendrobium candidumWall.ex Lindl. is also of this kind. However, both of them are short of resources. For a long time, there are no goods in the market. Now the domestic and foreign markets are called "Huoshan "Dendrobium", "Huodou", and "Jinhuodou" maple bucket products are not processed by the former two, but mostly copper-skinned Dendrobium produced in Huoshan (also known as Dendrobium ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 邓辉李耀亭陈乃富刘明珍
Owner 陈乃富
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