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ITS (Internal Transcribed Spacer) sequence and method for identifying certified lepidium meyenii walp and counterfeit lepidium meyenii walp as well as doped lepidium meyenii walp

A technology of maca and counterfeit products, applied in the field of identification of plant germplasm resources, can solve the problems of affecting product content, high price, multiple standards, etc.

Active Publication Date: 2014-07-02
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is extremely difficult to identify Maca dry slices and Maca powder from the aspects of character, smell and taste; the identification method of Maca secondary metabolites is analyzed by gas phase, liquid phase and mass spectrometry. On the one hand, it is expensive, and Secondary metabolites will affect the content of product ingredients to a certain extent due to different plant growth stages and environmental influences, which may lead to multiple standards, etc.

Method used

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  • ITS (Internal Transcribed Spacer) sequence and method for identifying certified lepidium meyenii walp and counterfeit lepidium meyenii walp as well as doped lepidium meyenii walp
  • ITS (Internal Transcribed Spacer) sequence and method for identifying certified lepidium meyenii walp and counterfeit lepidium meyenii walp as well as doped lepidium meyenii walp
  • ITS (Internal Transcribed Spacer) sequence and method for identifying certified lepidium meyenii walp and counterfeit lepidium meyenii walp as well as doped lepidium meyenii walp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] This embodiment has determined the amplifiability of ITS universal primers ITS4 and ITS5 to the yellow, red and black maca produced in Lijiang, the maca produced in Peru, the turnip produced in Lijiang, the rDNA ITS sequences of commercially available radish, corn and potato, and Comparing the full ITS sequences of these 8 materials, it shows that the Maca ITS region is highly conserved and has great differences from the ITS regions of turnip, radish, corn and potato. The full ITS sequence of Maca can be used as a commodity for Maca Barcodes are used to distinguish authentic Maca from counterfeit Maca. The method adopted includes the following specific steps:

[0019] (1) Extraction of total DNA: take dried yellow, red and black maca slices from Lijiang, maca powder from Peru, dried turnip slices, commercially available radish leaves, commercially available corn leaves and commercially available potato tubers, Use the Genestar kit to extract total DNA. The specific meth...

Embodiment 2

[0043] This embodiment utilizes the whole sequence of Maca ITS to identify the dry powder of Maca mixed with 50% potato powder, and the method adopted includes the following specific steps:

[0044] (1) Extraction of total DNA: Take about 2 g of maca dry powder mixed with 50% potato powder, and use the Genstar kit to extract total DNA. The specific method is as follows:

[0045] ① Add 700 μL of solution A in the kit, shake gently for 10 seconds, place in a water bath at 65°C for 30 minutes, and shake once every 15 minutes;

[0046] ② Add 700 μL of chloroform:isoamyl alcohol (24:1) mixture and shake vigorously for 10 seconds, then centrifuge at 12000 rpm for 10 minutes;

[0047] ③ Cut off the tip of the 200μL pipette tip, carefully absorb the supernatant and transfer it into a new 1.5mL centrifuge tube;

[0048] ④ Add twice the volume of pre-cooled absolute ethanol, shake slowly and place in a -20°C refrigerator for 20 minutes;

[0049] ⑤Pick flocculent DNA samples with a pip...

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Abstract

The invention relates to an ITS (Internal Transcribed Spacer) sequence and a method for identifying certified lepidium meyenii walp and counterfeit lepidium meyenii walp as well as doped lepidium meyenii walp. The invention discloses the r DANITS complete sequence of the certified lepidium meyenii walp and a method for identifying the certified lepidium meyenii walp and the counterfeit lepidium meyenii walp by using the sequence. The method comprises the following steps of: extracting a total DNA, amplifying by using an ITS region primer, recycling purified products, connecting carriers and screening positive clones, sequencing, carrying out sequence alignment, and distinguishing the authenticity of the lepidium meyenii walp. According to the method, the authenticity of the lepidium meyenii walp and whether other possible counterfeits such as turnips, radishes, corns, potatoes and the like are doped in lepidium meyenii walp powder can be effectively detected, and the certified lepidium meyenii walp and the counterfeit lepidium meyenii walp as well as the doped lepidium meyenii walp can be reliably, stably and efficiently identified.

Description

technical field [0001] The invention belongs to the technical field of identification of plant germplasm resources, and in particular relates to a nucleotide sequence and identification method for the authenticity identification of Maca, a new resource food plant. Background technique [0002] Maca (Lepidium meyenii Walp.) is an annual herbaceous plant belonging to the genus Brassicaceae. It is native to the Andes Mountains at an altitude of 3500-4500m in South America. Maca is rich in nutritional value, and the local Incas used it as food. In addition, Maca can also improve fertility and is used to solve the problem of human and animal infertility in the deserted and cold highlands. With people's understanding of Maca's nutritional value and medicinal value, the demand for Maca is also increasing worldwide. At present, the United States, Japan, Spain, Ecuador, Bolivia, Australia, etc. have successively planted, but have not formed a scale. my country began to introduce a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 赵兵陈金金王丽卫赵庆生王晓东袁晓凡
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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