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Method for identifying variety of glechoma longituba by utilizing ITS2 sequence

A Huoxuedan and sequence technology, applied in biochemical equipment and methods, special data processing applications, microbial measurement/inspection, etc., can solve the problem of low accuracy of identification methods and achieve the effect of improving accuracy

Active Publication Date: 2018-12-07
XIAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for identifying Huoxuedan varieties using the ITS2 sequence, which solves the problem of low accuracy of existing identification methods

Method used

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  • Method for identifying variety of glechoma longituba by utilizing ITS2 sequence
  • Method for identifying variety of glechoma longituba by utilizing ITS2 sequence
  • Method for identifying variety of glechoma longituba by utilizing ITS2 sequence

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Embodiment Construction

[0038] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0039] 1. Sample source, see Table 1 for details.

[0040] Table 1 Sample source

[0041]

[0042]

[0043] 2. DNA extraction of the sample to be tested

[0044] Step 1.1, pretreatment of Huoxuedan samples to be identified

[0045] Take 3 dried leaves of Huoxuedan to be identified in a centrifuge tube with a volume of 1.5mL, and place them in liquid nitrogen for quick freezing, then take out the centrifuge tube, and place the dried leaves of Huoxuedan to be identified in a plastic foam container containing liquid nitrogen , ground into powder with a small stick;

[0046] Step 1.2, after the temperature of the powdery dry leaves rises to room temperature, add 550uL of CTAB to the plastic foam container in step 1.1, heat in a dry bath at 65°C for at least 30 minutes, invert and mix well during the heating period, then place it for 10 m...

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Abstract

The invention discloses a method for identifying variety of glechoma longituba by utilizing an ITS2 sequence. The method comprises the following steps: 1, taking a plurality of dry leaves of the to-be-identified glechoma longituba, and extracting DNA of samples; 2, performing PCR amplification on the ITS2 nucleotide sequence; 3, performing bidirectional sequencing on a PCR product obtained in thestep 2 through a sequencing instrument to obtain a sequencing result, and performing sequence splicing, annotation and comparison on the sequencing result to obtain rDNA ITS2 sequences; and 4, building a phylogenetic tree based on the obtained rDNA ITS2 sequences and identifying the variety of the glechoma longituba. According to the method for identifying the variety of the glechoma longituba disclosed by the invention, through improvement of a DNA extraction method of dry medicinal materials and special PCR amplification reaction conditions, a molecular identification evidence is provided for identifying the glechoma longituba medicinal materials, the glechoma longituba and easily-confused products of the glechoma longituba can be accurately and quickly detected from the multiple traditional Chinese medicines, the accuracy of detection is improved, and the method is suitable for wide popularization and application.

Description

technical field [0001] The invention belongs to the technical field of plant variety identification methods, and in particular relates to a method for identifying Huoxuedan varieties by using ITS2 sequences. Background technique [0002] Glechoma longituba (Nakai) Kupr. is a perennial herbaceous plant of the Lamiaceae, also known as Lianqiancao, Diqianer, Qiandiqian, Penetrating Wind, Crossing the Wall Wind, Jiulixiang, etc., with a stem height of 10-20cm. Quadrilateral; leaves opposite, round or kidney-shaped; cymes axillary, with 2-6 flowers in each round; nutlets, fruiting period 4-5 months. Huoxue Dan likes dampness, and grows in grasslands, riversides or fields in forests. Its dry aerial part is used as medicine. The medicinal material is Lianqiancao (collected in "Chinese Pharmacopoeia" (2010 edition)), mainly produced in Jiangsu and Zhejiang. It is slightly cold in nature and pungent in taste. , inducing diuresis to relieve stranguria, dissipating blood stasis and d...

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Application Information

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IPC IPC(8): C12Q1/6869G06F19/24
CPCC12Q1/6869C12Q2531/113
Inventor 郑梦迪张彦张寒
Owner XIAN MEDICAL UNIV