DNA molecule identifying method for bluish dogbane and poacynum hendersonii

A technology of DNA molecules and large-leaf white hemp is applied in the field of DNA molecular identification of apocynum and large-leaf white hemp, and can solve the problems of considerable difficulty in accurate identification, no literature reports, and high repeatability.

Inactive Publication Date: 2007-04-11
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] There are many identification methods for plant Chinese herbal medicines and their counterfeit products, such as identification from morphology, identification from unique substances, identification from specific chemical reactions, identification by fingerprints, etc., but these identification methods are not High repeatability without identification by genetic analysis, not affected by environmental and human conditions
Predecessors have done some identification research on the morphology, microscopic characteristics and effective chemical components of the leaves of Apocynum and white hemp. It exists on both sides with the same density, the microscopic properties of the stomata are very similar, and the texture of the leaves is herbaceous; and when the leaves of Apocynum apocynum are used for medicine and tea, they have to be further processed, which increases the difficulty of morphological identification; predecessors The types and contents of flavonoids in two kinds of "Apocynum" leaves were also compared and studied, but it was found that the main chemical components contained in them were similar, and the chemical components and contents were easily affected by the environment, individual development stage, tissues and organs, etc. Therefore, it is more feasible to use genetic analysis method to identify Apocynum and Dayebaba
At present, there is no literature report on the genetic analysis and identification methods of Apocynum cylindrica and Dayebaba

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 47 samples of apocynum from 12 provinces including Xinjiang, Inner Mongolia, Henan, and Jilin, and 17 samples of big-leaf white hemp from three provinces including Xinjiang, Gansu, and Qinghai (all passed expert identification), after first extracting genomic DNA; Use primers SEQ ID No.1 and primers SEQ ID No.2 to carry out PCR amplification reaction on the extracted genomic DNA. The specific procedure is: 95°C pre-denaturation for 5 minutes, 94°C for 1 minute, 50-62°C for 1 minute, 72°C Extend for 1 min, and after 30 cycles, extend for 7 min at 72°C. PCR products were detected by conventional agarose gel electrophoresis, and all samples were found to have a clear single band at 220bp.

[0028] Further use primers SEQ ID No.1 and primers SEQ ID No.2 to continue the second round of PCR reaction on the extracted genomic DNA. The specific procedures are: 95°C pre-denaturation for 5 minutes, 94°C denaturation for 1 minute, 62.7°C renaturation for 1 minute, 72°C Extended at...

Embodiment 2

[0030] Four samples of two kinds of Apocynum tea and two dried leaves of Apocynum were purchased from the market respectively. Genomic DNA was extracted from each sample separately. First, use the specific primers SEQ ID No.1 and SEQ ID No.2 to carry out the first round of PCR amplification reaction. The specific reaction procedure is: pre-denaturation at 95°C for 5 min, denaturation at 94°C for 1 min, renaturation at 50-62°C for 1 min, and 72°C. Extended at ℃ for 1 min, and after 30 cycles, extended at 72°C for 7 min. After the PCR reaction was completed, conventional agarose electrophoresis was performed, and it was found that there was a clear single band at 220 bp in the electrophoresis pattern. It shows that the 4 samples to be detected are all Apocynum or Pleurotus chinensis, not other plants.

[0031]Progressively use primers SEQ ID No.1 and primers SEQ ID No.2 to continue the second round of PCR reaction on the extracted genomic DNA. The specific procedures are: pre-...

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Abstract

The present invention discloses DNA molecule identifying method of bluish dogbane and poacynum handersonii. The method includes the steps of extracting the genome DNA of bluish dogbane and poacynum handersonii separately, twice PCR proliferation and agarose gel electrophoresis in two different annealing temperatures with the extracted genome DNA as template and the specific primer, and identifying bluish dogbane and poacynum handersonii based on the electrophoresis behaviors of the PCR products. The method is not affected by environment factors, is independent on individual growth stages, and has no tissue and organ specificity, high repeatability, high stability and other advantages.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a DNA molecular identification method of apocynum and white hemp. Background technique [0002] Apocynum is a perennial herb of the genus Apocynum in the family Apocynaceae. It has been used as medicine for more than a thousand years. According to the "Dictionary of Chinese Medicine", Apocynum contains flavonoids, cardiac glycosides, amino acids and other medicinal ingredients. It has a good effect on the prevention and alleviation of hypertension, hyperlipidemia, coronary heart disease, asthma, bronchitis and other diseases; Apocynum leaves are also used for tea and cigarettes, according to "The Pharmacopoeia of the People's Republic of China" and "Compendium of Materia Medica" According to records, Apocynum tea has functions such as calming the liver and calming the nerves, clearing away heat and diuresis, stopping dizziness, reducing inflammation and relieving cough, strengt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陆长梅张卫明顾龚平彭雪梅
Owner NANJING NORMAL UNIVERSITY
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