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Method and application for identifyingtaxonomic statusand species of pharbitis choisy and closely related genera

A technology of a closely related genus, Petunia, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as experimental design and methodological defects, contradictory conclusions, and inability to determine taxonomic status and species, etc., to achieve Reliable and accurate identification results, the effect of accurate identification results

Active Publication Date: 2018-01-09
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Although some domestic scholars have done some exploratory studies on molecular science, there are defects in experimental design and methodology, and the conclusions obtained are inconsistent with "Flora of China" or "Chinese Pharmacopoeia", so they cannot be used as reference standards
At present, the existing research is generally one species and one sample. Such a sampling method ignores the relationship between the intra-species genetic diversity and the inter-species genetic diversity, and the results obtained can only determine the close relationship between species. Its taxonomic status and species cannot be determined, and its species identification can only be established when the intra-species diversity is less than the inter-species diversity

Method used

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  • Method and application for identifyingtaxonomic statusand species of pharbitis choisy and closely related genera
  • Method and application for identifyingtaxonomic statusand species of pharbitis choisy and closely related genera
  • Method and application for identifyingtaxonomic statusand species of pharbitis choisy and closely related genera

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Embodiment 1

[0054] Identification methods for the taxonomic status and species of Pharbitis hederacea Jacq., Pharbitis nil(L.) Roth. and Round-leaf morning glory (Pharbitis purpurea(L.) Roth):

[0055] 1. Plants to be tested: a total of 64 plants to be tested, including 7 split-leaf morning glory, 21 round-leaf morning glory, and 18 round-leaf morning glory.

[0056] Experimental equipment: bead mill tissue grinder, desktop high-speed centrifuge, PCR instrument, electrophoresis instrument, gel imaging analysis system.

[0057] 2. DNA extraction: Take 20mg of dried silica gel leaves or 1-2 morning glory seeds into a 2mL centrifuge tube, and use a bead-type tissue grinder to crush for 40 seconds until the material becomes powdery. Then use the plant genomic DNA extraction kit for DNA extraction. For specific methods, refer to the instructions of the DNA extraction kit.

[0058] 3. PCR amplification:

[0059] (1) Reaction system: 25uL, that is, 10×buffer (containing Mg 2+ ), 2.5uL; 2.5mM ...

Embodiment 2

[0074] The identification method of the Chinese herbal medicine Petunias: the plants to be tested include the Chinese medicinal materials to be tested and Petunia, and the rest of the steps and judging principles are consistent with the steps 2-8 described in Example 1. The specific judgment results are as follows: in the phylogenetic trees obtained respectively based on the rbcL gene and the ITS gene as primers of the Chinese medicinal material to be tested and the Fructus Petuniae, it is judged whether the Chinese medicinal material to be tested and the Fructus Petuniae are respectively aggregated into a monophyletic, and if so, it is judged that the Chinese medicinal material to be tested is not Belongs to the morning glory, if not, it is judged that the Chinese medicinal material to be tested belongs to the morning glory.

[0075] Calculate the sequence matrix obtained after the amplified rbcL gene and ITS gene of the Chinese herbal medicine to be tested and Petunia chinens...

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Abstract

The invention belongs to the technical field of biological detection and in particular relates to a method and an application for identifying taxonomic status and species of pharbitis choisy and closely related genera. The method comprises the following steps: extracting genome DNA of to-be-detected plants to obtain a sequence matrix; establishing an evolutionary tree based on the sequence matrix;establishing DNA bar codes based on the sequence matrix, respectively taking the evolutionary tree and the DNA bar codes to perform result judgment, thereby obtaining the identification result, namely the results of the taxonomic status and species of the identified to-be-detected plants if the judgment results are consistent. According to the method disclosed by the invention, a standard methodis established to accurately determine the taxonomic status and species of pharbitis choisy and closely related genera. According to the identification method, chloroplast gene rbcL and karyogene ITSareused for independent analysis, so that the obtained results can be confirmed with each other, the experiment conclusion is reliable, and the application range is wide.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a method and application for identifying the taxonomic status and species of the genus Petunia and related genera. Background technique [0002] Petunia is a genus under the family Convolvulaceae, which is an annual or perennial twining herb. There are about 24 species in this genus, which are widely distributed in temperate and subtropical regions. There are 3 species in our country, morning glory, color-changing morning glory, and round leaf morning glory, which are produced in both the north and the south. For the related plants of the genus Petunia and related genera, there have always been plants of this undetermined taxonomic status and species in the prior art, for example, for the three common "petunias" in China, namely the split-leaf morning glory (Pharbitis hederacea Jacq .), morning glory (Pharbitis nil (L.) Roth.) and round-leaved morning g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895
Inventor 邵秀玲张伟范晓虹吴兴海厉燕张京宣宋涛王简
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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