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SNP locus based CAPs molecular marker for identifying single seeds of amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola and applications of molecular marker

A molecular marker, Amaranthus amaranthus technology, applied in the field of molecular biology, can solve the problems of hindering application, long detection time, high cost and high cost, and achieve the effect of accurate and effective identification method, accurate detection and identification, and reliable detection method.

Active Publication Date: 2020-08-11
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many methods for SNP detection, but many methods are hindered in application due to high technical difficulty, high cost, and long detection time.

Method used

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  • SNP locus based CAPs molecular marker for identifying single seeds of amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola and applications of molecular marker
  • SNP locus based CAPs molecular marker for identifying single seeds of amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola and applications of molecular marker
  • SNP locus based CAPs molecular marker for identifying single seeds of amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola and applications of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 SNP sites and acquisition of CAPs molecular markers based on SNP sites

[0041] By analyzing and comparing the sequences of 18S, ITS, 26S and other genes of 52 samples from 18 species of Amaranthus L., and analyzing their phylogenetic relationships and sequence differences, the results were used to distinguish between amaranth and amaranth The SNP sites of Amaranthus sylvestris and other plants of the genus Amaranth, wherein the SNP site used to distinguish Amaranthus sylvestris from other plants of the genus Amaranth is located at position 233 of the sequence shown in SEQ ID NO.1. This position is A, and that of other amaranth plants is C. The SNP site used to distinguish Amaranthus squarrosa / Amaranthus from other amaranths is located at the 200th position of the sequence shown in SEQ ID NO.2. The position of Amaranthus squarrosa and Amaranthus sphaerocarpa is C, and other amaranths This site in plants is A / T.

[0042] The specific names and sources of the 52 pl...

Embodiment 2

[0060] Example 2 Method for distinguishing and identifying single seeds of Amaranthus longifolia, Amaranthus scabra, Amaranthus dulcis and other amaranth plants

[0061] The CAPs molecular markers provided in Example 1 and their specific detection primers were used to identify Amaranthus longifolia, Amaranthus scabra, and Amaranthus sphaerocephala. The specific methods are as follows:

[0062] 1. DNA extraction

[0063] Grind the single seed of the amaranth plant to be tested and place it in a 1.5ml centrifuge tube, use Lysase A in a water bath at 95°C for 10 minutes, and centrifuge for 2 minutes for later use.

[0064] 2. PCR amplification

[0065] (1) Amplification of CAPs molecular marker shown in SEQ ID NO.1

[0066] Using the DNA obtained in step 1 as a template, the forward primer 78F: 5'-GTCCCTGCCCTTTGTACACA-3' and the reverse primer 419R: 5'-GGGTTCGTTGTTGCATCGAG-3' were used for PCR amplification.

[0067] The PCR reaction system in 50μl is: 30ng / μl template DNA 1μl, 10pmol / μl pri...

Embodiment 3

[0080] Example 3 Discrimination and identification of Amaranthus longifolia, Amaranthus scabra and Amaranthus sylvestris

[0081] The single seeds of 52 plant material samples shown in Table 1 were used as the samples to be tested, and the method provided in Example 2 for distinguishing and identifying Amaranthus serrata, Amaranthus scabra, Amaranthus sylvestris, and other amaranths was used to perform Amaranthus spp. Discrimination and identification of seeds of Amaranthus scaber and Amaranthus sphaerocephala.

[0082] A mixed sample of single seeds of different plant materials in 52 plant material samples shown in Table 1 was used as a control.

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Abstract

The invention relates to the technical field of molecular biology, and specifically relates to a SNP locus based CAPs molecular marker for identifying single seeds of amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola and applications of the molecular marker. The CAPs molecular marker is shown as SEQ ID NO. 1 and SEQ ID NO. 2, and specific detection primers are shown as SEQ ID NO. 3-6. A PCR-RFLP method used for distinguishing and identifying the single seeds of amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola and other amaranthus seeds is provided. Through the utilization of the provided CAPs molecular marker and the PCR-RFLP method, the rapid and accurate detection identification of the single seeds of the amaranthus palmeri, amaranthus tuberculatus and amaranthus aeronicola can be realized; and a reliable detection method can be provided for the rapid identification of quarantine weeds of the amaranthus palmeri, the amaranthus tuberculatus andthe amaranthus aeronicola at ports.

Description

Technical field [0001] The present invention relates to the technical field of molecular biology, in particular to CAPs molecular markers based on SNP sites for identifying single seeds of Amaranthus longifolia, Amaranthus scabra and Amaranthus serrata and its application. Background technique [0002] The quarantine weeds Amaranthus longifolia, Amaranthus scabra, and Amaranthus dulcis belong to the subgenus Amaranthus heterophyllus of the family Amaranthaceae. They are native to North America and are super weeds in soybean and corn farms in the United States. The seeds of Amaranth are small, only about 1mm. The morphology of seeds of plants of the same genus is similar, and it is difficult to distinguish effectively according to the morphology of seeds. Therefore, the development of a molecular biological detection method that can quickly and efficiently identify the single seeds of Amaranthus serrata, Amaranthus scabra and Amaranthus serrata is of great significance for port de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/683C12N15/11
CPCC12Q1/6895C12Q1/683C12Q2600/13C12Q2600/156C12Q2531/113
Inventor 徐晗
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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