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Method for identifying age limit of ginseng by using telomere length of ginseng

A ginseng and aging technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of mechanical damage, difficult observation of reed bowls, difficulty in ensuring the integrity of ginseng shape, etc., to achieve less sample volume and ginseng damage small effect

Active Publication Date: 2015-01-21
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional method of identifying the age of ginseng is character identification, that is, judging the age of ginseng by observing the number of reed bowls of ginseng. The way to increase the number of reed bowls is shoddy
Chemical analysis is also often used as one of the methods to identify the age of ginseng, but this method often requires several grams of ginseng powder samples, which is difficult to ensure the integrity of ginseng shape, which greatly damages the value of ginseng

Method used

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  • Method for identifying age limit of ginseng by using telomere length of ginseng
  • Method for identifying age limit of ginseng by using telomere length of ginseng
  • Method for identifying age limit of ginseng by using telomere length of ginseng

Examples

Experimental program
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Embodiment 1

[0032] Example 1. Establishment of a method for identifying the age of ginseng by using ginseng telomere length

[0033] Ginseng tested: ginseng samples of different years from two regions, see Table 1 for details.

[0034] Table 1 Ginseng samples of different years in two regions

[0035]

[0036] 1. Extraction of Ginseng Genomic DNA

[0037] Genomic DNA of each tested ginseng was extracted by CTAB method. details as follows:

[0038] Take 0.1 g dry weight (no mildew) of the ginseng to be tested located 1-2 cm below the reed head, put it in a pulverizer, grind it, and pass it through a 40-mesh sieve. Transfer the powder to a 2.0mL Eppendorf tube, add 900μL of sterilized CTAB extract (recipe: 2% (2g / 100ml) CTAB, 100mmol / L Tris-HCl pH=8.0, 20mmol / LEDTA, 1.4mol / L NaCl), 0.02g PVP 40000, 10μL β-mercaptoethanol, fully vortexed and mixed, in a water bath at 65°C for 1.5h-2h, and gently shaked 2-3 times during the period. After completion, take it out and cool to room temper...

Embodiment 2

[0056] Example 2, the application of the method of identifying the age of ginseng by using the telomere length of ginseng

[0057] In order to further verify the accuracy of the method established in Example 1 to identify the age of ginseng by using the telomere length of ginseng, the inventors of the present invention collected cultivated ginseng samples of different ages, and conducted a double-blind experiment on the age prediction of ginseng using the evaluation formula established above .

[0058] Referring to steps 1 to 3 in Example 1, the telomere restriction fragment lengths were measured for each ginseng sample of known age. Substituting the measurement results into y=0.827x+8.231, the obtained x value is the predicted years.

[0059] The results are shown in Table 3, which shows that the prediction of ginseng age using this formula is close to the actual age.

[0060] Table 3 Prediction results of ginseng age

[0061]

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Abstract

The invention discloses a method for identifying the age limit of a ginseng by using the telomere length of the ginseng. The method comprises the following steps: 1) taking a sample, 1-2 cm below a fibrous root, of a to-be-detected ginseng, and extracting total DNA; 2) taking the total DNA, and carrying out digestion on the total DNA by using an incision enzyme which can recognize and cut double-stranded DNA shown in 5'-TCGA-3'; 3) carrying out Southern blotting hybridization or dot hybridization on a digestion product by taking a single-stranded DNA shown in 5'-CCCTAAA-3' as a probe; 4) according to a hybridization result, measuring the length of a restriction fragment of a hybridization product so as to obtain the length of the telomere restriction fragment of the ginseng, and taking the length as to-be-detected TRF; and 5) by taking the to-be-detected TRF as a y value, substituting the to-be-detected TRF into a formula y = 0.827x + 8.231, so that an obtained x value is the age limit of the to-be-detected ginseng. Experiments show that the goodness of fit between an age limit of a ginseng measured by using the method and an actual life of the ginseng is extremely high; and the required sample amount in the method is extremely few, so that the method has extremely minimal damages on the ginseng, and therefore, the method is a nondestructive identifying method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for identifying the age of ginseng by utilizing the telomere length of ginseng, in particular to a method for identifying the age of ginseng by using the length of the telomere restriction fragment (TRF value) of ginseng. Background technique [0002] Ginseng, the root of the Araliaceae plant, has been used as a tonic and strong medicine in traditional Chinese medicine for nearly a thousand years. Ginseng blooms in the fourth year after planting, and its roots mature gradually in 4-6 years and can survive for hundreds of years. Studies have shown that the longer the age of ginseng roots, the higher the content of ginsenosides, making it more medicinal and commercial. [0003] The traditional method of identifying the age of ginseng is character identification, that is, judging the age of ginseng by observing the number of reed bowls of ginseng. The way to increase the number...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6816C12Q2521/301
Inventor 黄璐琦蒋超袁媛王尧龙
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI