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SNP-based caps molecular markers for identification of single seeds of Palmer amaranthus, Amaranth amaranthus and Amaranthus sativus and its application

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 identification, and reliable detection method.

Active Publication Date: 2022-06-07
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-based caps molecular markers for identification of single seeds of Palmer amaranthus, Amaranth amaranthus and Amaranthus sativus and its application
  • SNP-based caps molecular markers for identification of single seeds of Palmer amaranthus, Amaranth amaranthus and Amaranthus sativus and its application
  • SNP-based caps molecular markers for identification of single seeds of Palmer amaranthus, Amaranth amaranthus and Amaranthus sativus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 SNP site and acquisition of SNP site-based CAPs molecular markers

[0041] By analyzing and aligning the sequences of 18S, ITS, 26S and other genes in 52 samples of 18 species of Amaranthus L., and analyzing their phylogenetic relationships and sequence differences, the results obtained for the distinction between Amaranthus amaranthus and Amaranthus amaranthus , SNP sites of Amaranthus serrata and other Amaranthus plants, wherein, the SNP site used to distinguish Amaranthus from other Amaranth plants is located at position 233 of the sequence shown in SEQ ID NO. This site is A, and this site is C in other Amaranth species. The SNP site for distinguishing Amaranthus amaranthus / Amaranthus from other amaranth plants is located at the 200th position of the sequence shown in SEQ ID NO. This site in plants is A / T.

[0042] The specific names and material sources of the 52 plant materials used for gene amplification and sequence analysis are shown in Table 1, and ...

Embodiment 2

[0060] Embodiment 2 The method for distinguishing and identifying the single seed of Amaranthus amaranthus, Amaranthus chinensis, Amaranthus serrata and other Amaranth plants

[0061] Utilize the CAPs molecular markers provided in Example 1 and their specific detection primers to identify Amaranthus amarantensis, Amaranth amaranthus, Amaranthus serrata, and the specific methods are as follows:

[0062] 1. DNA extraction

[0063] The single seed of the amaranth plant to be tested was ground and placed in a 1.5 ml centrifuge tube, and was then used in a water bath at 95° C. for 10 minutes with Lysase A, and then centrifuged for 2 minutes for use.

[0064] 2. PCR amplification

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

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

[0067] The PCR reaction system is cal...

Embodiment 3

[0080] Embodiment 3 Differentiation and identification of Amaranthus amaranthus, Amaranthus amaranthus, Amaranthus serrata

[0081] Taking the single seeds of 52 plant material samples shown in Table 1 as the sample to be tested, the method for distinguishing and identifying Amaranth, Amaranth serrata, Amaranth and other Amaranth plants provided in Example 2 was used to carry out Amaranth. , Distinctive identification of Amaranthus amaranthus and Amaranthus amaranthus seeds.

[0082] A mixed sample of single seeds of different plant materials in the 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, in particular to CAPs molecular markers based on SNP sites for identifying single seeds of palmer amaranth, amaranth and amaranth and application thereof. The CAPs molecular marker of the present invention is shown in SEQ ID NO.1 and SEQ ID NO.2, and its specific detection primer is shown in SEQ ID NO.3‑6. The invention provides a PCR-RFLP method for distinguishing and identifying the seeds of palmer amaranth, amaranth, and amaranth from other amaranthus seeds. Utilize the CAPs molecular marker provided by the present invention and the PCR-RFLP method can realize rapid and accurate detection and identification to the single seed of Palmer amaranth, Amaranthus amaranthus, and Amaranthus saponifera, and quickly identify quarantine weeds Palmer amaranth, Amaranth Amaranth and amaranthus provide reliable detection methods.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a SNP site-based CAPs molecular marker for identifying single seeds of Amaranthus lanceolata, Amaranthus serrata and Amaranthus serrata and application thereof. Background technique [0002] Quarantine weeds Amaranthus amaranthus, Amaranthus amaranthus and Amaranthus amaranthus belong to the subgenus Amaranthus in the family Amaranthaceae. They are native to North America and are super weeds in soybean and corn fields in the United States. The seeds of amaranth are small, only about 1mm, and the seeds of different species of the same genus are similar in shape, so it is difficult to effectively distinguish according to the seed shape. Therefore, it is of great significance for port detection to develop a molecular biological detection method that can quickly and efficiently identify single seeds of Amaranthus amaranthus, Amaranthus chinensis and Amaranthus serrata. [...

Claims

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Application Information

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Patent Type & Authority Patents(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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