Method for identifying zygophyllum xanthoxylum

A kind of overlord and species technology, applied in the field of plant species identification, can solve the problems of the limitation of overlord identification, the distinction of unfavorable varieties, and the impact on the accuracy of identification, and achieve the effect of rapid identification, easy operation and strong practicability

Inactive Publication Date: 2019-04-05
CHINESE RES ACAD OF ENVIRONMENTAL SCI
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Problems solved by technology

[0004] At present, the variety identification of Bawang is limited to traditional morphological analysis, microstructure analysis, chemical fingerprint, etc., but these traditional identification methods have their own shortcomings
For xerophytic species with similar shapes, it is difficult to distinguish through trait identification, and plant morphology is easily affected by many factors such as geographical environment and growth period, thus affecting the accuracy of identification; chemical analysis methods based on the determination of active ingredient content are easily affected by their physiological Conditions, storage and other factors are not conducive to the distinction of varieties; at the same time, HPLC, CE, MS and other chemical analysis instruments and equipment are expensive and cannot be afforded by ordinary laboratories
Relying on traditional morphological taxonomy methods has limitations in the identification of Bawang

Method used

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  • Method for identifying zygophyllum xanthoxylum
  • Method for identifying zygophyllum xanthoxylum
  • Method for identifying zygophyllum xanthoxylum

Examples

Experimental program
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Embodiment 1

[0027] This embodiment has determined the ribosomal DNA ITS sequence from 9 place of origin Bawang (Paragon, Bayin Wendur, Zongbieli, Chaogetu Huresumu, Bayannuorigong, Xini Town, Baiyin Engel, Zhuozhuoshan, Mu Us), the steps are as follows:

[0028] 1. Sample collection and processing

[0029]The mature Bawang leaves (collected locally) of the current year were used as the material for extracting genomic DNA, and they were dried in a drying box filled with silica gel.

[0030] 2. Extraction of total genomic DNA

[0031] The kit (EE111-12) provided by Beijing Quanshijin Biotechnology Co., Ltd. was used

[0032] ①Place EB in a water bath (60-70°C) for constant temperature preheating;

[0033] ② Cut a small amount of sample (approximately 30 mg of dried tissue) into a 2ml centrifuge tube, put several small steel balls into the centrifuge tube at the same time, place the centrifuge tube in a plant high-speed tissue grinder for full grinding, centrifuge program Set 800rpm, 50s...

Embodiment 2

[0061] The difference from Example 1 is only that the contrast plants are replaced with pearl pigtail, red sand, quadrangle wood, caragana, and holly. The ITS sequences of the above species obtained through the NCBI database were used as reference sequences for comparison and cluster analysis with the obtained Bawang ITS sequences. The sequence comparison results are as follows Figure 4 As shown, the homology rate of the sequence comparison is greater than 95%, which can be regarded as originating from the same species, so that different germplasm resources can be distinguished according to the comparison results, and the purpose of identifying the overlord can be achieved. Clustering results such as Figure 5 As shown, the homology rate of Bawang in 9 regions is high, and they are clustered into one branch, and other species are clustered into another branch, but the homology with Bawang is far away, so they can be distinguished from Bawang. In summary, the method of the pr...

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Abstract

The invention provides a method for identifying whether a sample is zygophyllum xanthoxylum or not. The method comprises the following steps that (1) the total DNA of a genome is extracted from the sample; (2) the DNA extracted in the first step is used as a template; a specific primer pair used for amplifying ITS sequences is used for PCR amplification; (3) a PCR product is sequenced; (4) the sequenced result is subjected to sequence comparison with ITS sequences of one or various known zygophyllum xanthoxylum species; (5) whether the sample belongs to the zygophyllum xanthoxylum or not according to the sequence comparison result. The invention also provides a relevant purpose and a kit.

Description

technical field [0001] The invention relates to the field of identification of plant species, in particular to a method for identifying whether a sample belongs to the overlord species, in particular to a method for identifying the overlord species by using an ITS sequence. technical background [0002] Overlord (Zygophyllum xanthoxylum) is a xerophytic succulent shrub of Tribulus terrestae [1] , mainly distributed in deserts and steppe desert areas [2] . It is one of the main dominant species and construction species of shrub vegetation in the western desert area of ​​China. It has strong stress resistance, great ecological plasticity, strong drought resistance, salt tolerance and sand collection ability, and plays an important role in maintaining desert ecological balance. effect [3-5] . In recent years, due to the deterioration of the natural ecological environment in the distribution area and excessive firewood harvesting and grazing by the surrounding farmers and he...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6869
CPCC12Q1/6869C12Q1/6895C12Q2531/113C12Q2535/122
Inventor 沈奇关潇全占军臧春鑫赵志平冯朝阳
Owner CHINESE RES ACAD OF ENVIRONMENTAL SCI
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