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ITS (Internal Transcribed Spacer) used for identification of lepidium meyenii and accurate identification method of lepidium meyenii

A sequence and maca technology, applied in biochemical equipment and methods, resistance to vector-borne diseases, measurement/inspection of microorganisms, etc., can solve problems such as interference from microbial contamination, achieve low cost, and facilitate large-scale promotion and use , the effect of simple operation

Active Publication Date: 2020-08-11
广东天葆堂健康科技产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The ITS region sequence of the ribosomal genome has advantages in plant identification due to its fast evolution rate, good primer versatility, large number of sequences included in the NCBI database, and rich genetic information, but it is easily interfered by microbial contamination. As a result, the use of ITS identification has certain limitations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Obtaining the sequence of the rbcL region of maca samples

[0021] 1. DNA extraction of samples to be tested

[0022] (1) Multiple maca samples were collected from different origins, as shown in Table 1.

[0023] Table 1: Maca sample collection information

[0024] serial number sample Sampling site collection place 1 Maca the root Yunnan 2 Maca the root Tibet 3 Maca the root sichuan

[0025] (2) Adopt CTAB method to carry out DNA extraction to the sample to be tested in Table 1, concrete operation is as follows:

[0026] 1) Soak the sample to be tested in 75% ethanol for 5 minutes, then take it out, and place it in a sterile environment to air dry naturally.

[0027] 2) Take 2g of the sample to be tested, add liquid nitrogen and grind it into powder, put it in a 10mL centrifuge tube, then add 4.5mL of 3×CTAB extract preheated at 65°C, 0.5mL of SDS and mix well, then bathe in water at 65°C for 1h, and Shake...

Embodiment 2

[0053] Compared with Example 1, the difference is that the Maca in Example 1 is prepared into a powder, and the powder is used as a sample to extract DNA for identification. As a result, the above-mentioned 8 bases can also be specifically detected point.

[0054] SEQ ID NO: 1

[0055] tatactcctg aatatgaaac caaggatact gatatcttgg cagcattccg agtaactcct

[0056] caacccggag ttccacctga agaagcaggg gctgcggtag ctgctgaatc ttctactggt

[0057] acatggacaa ctgtgtggac cgatgggctt accagccttg atcgttacaa aggacgatgc

[0058] taccacatcg agcccgttcc aggagaagaa agtcaattta ttgcgtatgt agcttaccca

[0059] ttagaccttt ttgaagaagg ttcggttatact aacatgttta cctcgattgt gggtaatgta

[0060] tttgggttca aagccctggc tgctctacgt ctagaggatc tgcgaatccc tcctgcttat

[0061] actaaaactt tccagggacc gcctcatggt atccaagttg aaagagataa attgaacaag

[0062] tatggacgtc ccctattagg atgtactatt aaaccaaaat tgggattatc tgcgaagaac

[0063] tatggtagag cagttt

[0064] SEQ ID NO: 2

[0065] atgtcaccacaaacagagactaaagc

[0066] SEQ ID ...

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PUM

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Abstract

The invention discloses an ITS (Internal Transcribed Spacer) used for identification of lepidium meyenii and an accurate identification method of lepidium meyenii. The nucleotide sequence of the rbcLITS of lepidium meyenii is SEQ ID NO:1. The identification method comprises steps as follows: extracting DNA of a to-be-detected sample as an amplification template; using downstream and upstream primers of the rbcL ITS to perform a PCR amplified reaction to obtain an rbcL ITS; sequencing the amplified product, matching and pairing sequences and judging whether the to-be-detected sample is lepidium meyenii by checking base information of 8 sites. Compared with methods for taking ITS as a basis for identifying lepidium meyenii in existing patents or documents, the method can effectively performmolecular identification on the lepidium meyenii on the level of molecular biology, the risk of possible fungal contamination of the ITS is also avoided, and the method has the advantages of being simple to operate, low in cost and the like and can rapid and accurate identification of lepidium meyenii as well as sibling species and powder thereof.

Description

【Technical field】 [0001] The invention relates to the technical field of identification of plant species, in particular to a region sequence for identification of maca and a method for its precise identification. 【Background technique】 [0002] Maca is the root of Maca (Lepidium meyenii), also known as Peruvian ginseng. It is native to the Andes Mountains above 4,000 meters in South America and is one of the main food sources for local residents. Studies have shown that maca has the functions of improving fertility, enhancing immunity, anti-fatigue, relieving menopausal syndrome, and anti-oxidation. Due to these special effects of maca, it is widely used in health care products and food industries at home and abroad. [0003] It is recorded in "Flora of China" (1987 edition) that there are about 150 species of plants in the genus Lepidium, which are widely distributed all over the world. There are 15 species and one variety in my country, which are distributed all over the c...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2537/165Y02A50/30
Inventor 谭贵良朱爽胡敏李雪雁李梅
Owner 广东天葆堂健康科技产业有限公司