Molecular identification method of Yunnan manyleaf Paris rhizome

A technology of molecular identification and Dianzhong building, which is applied in the measurement/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of undiscovered molecular identification, avoid the uncertainty of PCR amplification success, make identification easy, Ensure authentic results

Inactive Publication Date: 2014-01-01
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through two novelty searches in 2011 and 2012, no relevant reports on the DNA Barcode and molecular identification of the plant of Diana dianthus have been found at home and abroad.

Method used

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  • Molecular identification method of Yunnan manyleaf Paris rhizome
  • Molecular identification method of Yunnan manyleaf Paris rhizome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 The construction of the barcode of Dianzhonglou

[0030] 1. Collection of samples of Dianzhonglou

[0031] Dianzhonglou is mainly distributed in Northwest Yunnan, Northeastern Yunnan, and Central Yunnan. For these areas, the present invention collected samples from 24 main distribution areas of Dianzhonglou in the early stage (including some southwest and southeast samples) , see Table 1 for details, which excludes the variation of the sequence information loci between individuals of different origins of Chonglou, making the unique DNA barcode of Chonglou more authentic and effective. All the collected samples of Zhonghua were confirmed to be Zhonglou through plant morphological identification. In addition, other different species under the genus Chonglou were also collected to distinguish the sequence difference between the DNA barcode of Zhonglou Dianchi and other species.

[0032] 2. Extraction of sample DNA

[0033] Each sample was 0.5g, according to...

Embodiment 2

[0058] Example 2 Molecular identification method of Paris polyphylla

[0059](1) Extract the total DNA of the sample to be identified: extract the total DNA of the sample to be identified according to the conventional CTAB method (Doyle J. DNA protocols for plants-CTAB total DNA isolation. In: Hewitt GM, Johnston A, eds. Molecular techniques in taxonomy. Berlin: Springer, 1991, 283-293.). Take 0.5g of the sample to be tested, and extract the total DNA according to the conventional CTAB method. After the extracted total DNA sample is purified by RNase A, measure the optical density values ​​at 260nm and 280nm with a UV spectrophotometer. When the DNA purity reaches OD260 If the / OD280 is between 1.6-1.8, the total DNA concentration is diluted to 200ng / μl for step (2) PCR reaction. If the optical density values ​​at 260nm and 280nm are measured with a UV spectrophotometer, and the DNA purity does not reach OD260 / OD280 between 1.6-1.8, it indicates that the DNA purity is not eno...

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Abstract

The invention discloses a molecular identification method of Yunnan manyleaf Paris rhizome, belonging to the technical field of molecular markers. The method comprises the steps of performing PCR (polymerase chain reaction) on a sample to be identified to obtain an amplified product, sequencing the amplified product, then comparing with a barcode of the Yunnan manyleaf Paris rhizome, and judging that the sample to be identified is the Yunnan manyleaf Paris rhizome when the similarity between the sequence of the amplified product and a base sequence shown in the barcode of the Yunnan manyleaf Paris rhizome is more than 98.2%, the bases at sites 1-4 are CTGA, the base at the site 19 is deleted, the bases at the site 77 are all C, the bases at the site 138 are all C, the base at the site 792 is G, the bases at the sites 1110-1113 are GGCG, the base at the site 1116 is G and the bases at the sites 1119-1120 are CC. By the method disclosed by the invention, the technical problem that sensory evaluation or physical and chemical analysis and other methods are difficult to identify the authenticity of the Yunnan manyleaf Paris rhizome and different Paris plants under Paris can be overcome and the plants of the Yunnan manyleaf Paris rhizome can be effectively, simply, conveniently and quickly identified.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to the molecular identification technology of the polyphylla dianthus plant under the genus Chonglou by using the PCR technology. Background technique [0002] The genus Paris L. belongs to the Parideae family of Liliaceae. There are 28 species in the genus, all of which are perennial herbs distributed in tropical and temperate regions of Eurasia. There are 22 species in China (18 species are endemic to my country), and the distribution and diversification center of the genus is from the Yunnan-Guizhou Plateau to the Qionglai Mountains in Sichuan; among the provinces and regions in China, 16 species are produced in Yunnan, of which 6 species are endemic to Yunnan, and its species diversity It also ranks first in the country (Ji et al. Phylogeny and classification of Paris (Melanthiaceae) inferred from DNA sequence data [J]. Annals of Botany, 2006, 98: 245-256). [0003] Pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q1/6895C12Q2563/185
Inventor 刘涛杨生超谢世清年丽菊李玛王玲
Owner YUNNAN AGRICULTURAL UNIVERSITY
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