Identification methods and uses of seed plant species

An identification method and technology for gymnosperms, applied in the field of biological detection, can solve problems such as being too large, not considering the genetic distance between species, etc., and achieve the effects of high application value, high accuracy, and low coverage requirements.

Inactive Publication Date: 2019-12-06
SHANDONG UNIV
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Problems solved by technology

Previous similar studies did not consider the value of the genetic distance between species when identifying species, or when determining the genetic distance between species, only obtained the average value by pairwise comparison of species within the same genus, and the value was obviously too large

Method used

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  • Identification methods and uses of seed plant species
  • Identification methods and uses of seed plant species

Examples

Experimental program
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Effect test

Embodiment 1

[0049] A method for identifying a seed plant species, comprising the steps of:

[0050] Determining molecular cut-off values: according to e.g. figure 1 The group relationship shown in the "Angiosperm APG II Classification System (Revised Edition)" and all representative classes of gymnosperms except the monotype (Ginkgophyla), selected 7866 species, including 99 angiosperms families, 179 genera, and 99 families, so that it basically covers the diversity of angiosperms, and the genera of the selected species meet the species coverage of more than 75%; download the ITS2 gene sequence of the group from GenBank, or download the group The ITS sequence of the selected species is intercepted according to the GenBank annotation to obtain the ITS2 gene sequence; the ITS2 gene sequence matrix of the selected species is imported into the MEGA software to build an evolutionary tree in units of genus. Multiple sequences of the same species must be input adjacently. According to the princi...

Embodiment 2

[0059] Determining the molecular threshold is consistent with the steps described in Example 1, so it will not be repeated in this example.

[0060] Determining the species category: a method for identification of a suspected medicinal plant yew, the specific implementation steps include:

[0061] Step 1. Sequencing of the species to be tested: extract the DNA of the species to be tested, perform PCR amplification, use ITS universal primers ITS4 and ITS5B as PCR amplification primers, and perform DNA sequencing on the PCR products. The sequence results of the species to be tested are shown as SEQ ID No: Shown in 4: the sequence of ITS4 is shown in SEQ ID No: 5; the sequence of ITS4 is shown in SEQ ID No: 6;

[0062] Step 2. Sequence alignment: Obtain the GenBank sequence numbers AF259300, AF259292, and AF259291 of the target species Taxus chinensis from GenBank, wherein the sequence of the GenBank sequence number AF259300 is shown in SEQ ID No: 2, The sequences of GenBank seq...

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Abstract

The invention relates to an identification method and application of a seed plant species, comprising the following steps: selecting a seed plant species, obtaining the ITS2 gene sequence of the seed plant species, constructing an evolutionary tree based on the ITS2 gene sequence, and selecting multiple pairs of sisters in the evolutionary tree Species, calculate the genetic distance between each pair of sister species, conduct a significant difference test on the genetic distances of multiple groups belonging to the same species, select the genetic distance with no significant difference to calculate the average value to obtain the molecular cut-off value; calculate the test species and the target The genetic distance between species is compared with the molecular cut-off value of the phylum to which the species to be tested is compared to determine the species category. The method of the present invention only needs to compare the degree of divergence between the ITS2 gene sequence of the sample to be tested or the suspected species and the target species, which is simpler and easier to implement, with high accuracy and strong objectivity, and does not require a large number of samples of other species of the same genus as For reference, it has high application value.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to an identification method and application of a seed plant species. Background technique [0002] At present, the main method of species identification is morphological classification. However, because of its poor accuracy, strong subjectivity and low efficiency, it can no longer meet the requirements of today's species classification and identification. With the improvement of sequencing level and the reduction of sequencing cost, the role of DNA sequence in species identification is becoming more and more prominent. In 2003, Canadian zoologist Paul Hebert first proposed the concept of DNA barcoding (DNA barcoding). It uses a standard, sufficiently variable, easy-to-amplify and relatively short DNA fragment to achieve rapid, accurate and automated species identification by establishing a biological DNA barcode database. DNA barcoding technology is currently the most...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G16B30/10G16B25/10G16B50/00
CPCC12Q1/6895C12Q2600/156G16B25/00G16B30/00G16B50/00
Inventor 张伟赵宏
Owner SHANDONG UNIV
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