Method for identifying plants by using DNA characteristic sequence

A DNA molecule and plant technology, applied in the field of plant identification using DNA characteristic sequences, can solve the problems of long development cycle, identification success rate of less than 80%, and heavy workload.

Active Publication Date: 2021-08-31
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the identification success rate of this scheme at the species level is less than 80%, and the operation and analysis process of identifying species using multiple barcodes is very cumbersome (Li D Z, Gao L M, Li H T, et al. Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants [J]. Proceedings of the National Academy of Sciences, 2011.)
However, traditional SCAR markers are usually converted from RAPD, SRAP, SSR markers, etc. Using the above types of markers to convert to SCAR markers often faces problems such as long development cycle and heavy workload.
At present, there is no report on the use of DSS to develop markers for medicinal plant identification and for identification of medicinal plants

Method used

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  • Method for identifying plants by using DNA characteristic sequence
  • Method for identifying plants by using DNA characteristic sequence
  • Method for identifying plants by using DNA characteristic sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, the establishment of the method for identifying plants using DNA signature sequences

[0048] The inventor of the present invention has established a method for using DSS to identify plants through a large number of experiments, and the specific steps are as follows:

[0049] 1. Obtaining DSS

[0050] 1. Obtain 4356 chloroplast genome sequences of 3899 (plant) species from database download or analysis of large amounts of data.

[0051]3899个物种中具有代表性的500个分别为Solidago decurrens、Erycibeobtusifolia、Erycibeschmidtii、Syzygiumaromaticum、Panax ginseng、Illiciumverum、Murraya exotica、Murrayapaniculata、Senegalia catechu、Canavalia gladiata、Panax notoginseng、Saururus chinensis、Sparganiumstoloniferum、Berberis vernae、Berberis poiretii 、Berberis soulieana、Berberis wilsoniae、Zingiber officinale、Toxicodendron vernicifluum、Inula helenium、Bolbostemmapaniculatum、Pseudolarixamabilis、Smilax glabra、Callicarpa macrophylla、Sargentodoxa cuneata、Glycinemax、Gleditsia sinensis、Isatis ti...

Embodiment 2

[0069] Embodiment 2, the method for identifying Atractylodes macrocephala using DNA characteristic sequence

[0070] 1. Acquisition of Baizhu DSS

[0071] According to the method of step 1 in Example 1, Atractylodes macrocephala DSS was obtained.

[0072] A total of 1327 DSS items of Atractylodes macrocephala were obtained, among which the first 100 items are shown in Table 1.

[0073] Table 1

[0074]

[0075]

[0076]

[0077] 2. Randomly select 5 pieces from the 1327 pieces of Atractylodes macrocephala DSS obtained in step 1, and design and synthesize primer pairs for amplifying the DSS according to the upstream and downstream 200 bp fragments of their positions in the genome.

[0078] The five selected Atractylodes macrocephala DSSs, the five primer pairs (composed of upstream primers and downstream primers) used to amplify the DSSs, and the length of the amplification products are shown in Table 2.

[0079] Table 2

[0080]

[0081]

[0082] Note: prime...

Embodiment 3

[0095] Embodiment 3, the method for identifying ginseng using DNA signature sequence

[0096] 1. Obtaining Ginseng DSS

[0097] Ginseng DSS was obtained according to the method of step one in Example 1.

[0098] A total of 5,927 ginseng DSSs were obtained, of which the top 100 are shown in Table 4.

[0099] Table 4

[0100]

[0101]

[0102]

[0103] 2. Five ginseng DSSs were randomly selected from the 5927 ginseng DSSs obtained in step 1, and primer pairs for amplifying the DSSs were designed and synthesized according to the upstream and downstream 200 bp fragments of their positions in the genome.

[0104] The five selected ginseng DSSs, the five primer pairs (composed of upstream primers and downstream primers) used to amplify the DSSs, and the length of the amplified products are shown in Table 5.

[0105] table 5

[0106]

[0107] 3. Use the 5 primer pairs synthesized in step 2 to identify Atractylodes macrocephala

[0108] The samples to be tested were 8...

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Abstract

The invention discloses a method for identifying plants by using a DNA characteristic sequence. The method is realized by detecting whether a genome of a plant contains DSS of a target species or a reverse complementary sequence thereof; the DSS of the target species is a nucleic acid sequence fragment which has a length of 20-100bp, exists in a nucleic acid sequence of the target species and does not exist in a nucleic acid sequence of a non-target species. For example, bighead atractylodes rhizome DNA can be identified from other atractylis plants with extremely close genetic relationship, and the defect that only rhizoma atractylodis and bighead atractylodes rhizome can be distinguished in the prior art is overcome. The DSS is not limited to a specific region of the genome, the data source is rich, and the specificity is good. The method has important application value in identifying medicinal plants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for identifying plants using DNA characteristic sequences. Background technique [0002] DNA detection technology has gradually become the preferred technology for species identification. Appropriate mutation rate, development of sequencing technology and increasing nucleic acid data endow DNA detection technology with advantages. Among the existing DNA detection technologies, the DNA barcoding technology (DNA barcoding) proposed by Hebert et al. is currently one of the most widely used methods. DNA barcodes refer to standard, sufficiently variable, easy-to-amplify, and relatively short DNA fragments that can represent the species in an organism. In recent years, with the increase in the data volume of reference databases (such as Barcode of Life Data System, BOLD), the improvement of computer algorithms (such as OverlapPER), and the increase in related researc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 袁媛华中一蒋超黄璐琦
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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