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DNA identification method of Isodon serra(Maxim.)Kudo and Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara

A technology of Fragrant Tea Vegetables and Xihuangcao, which is applied in the field of plant species identification

Active Publication Date: 2015-03-11
GUANGZHOU BAIYUSN HUTCHISON WHAMPOA CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RAPD technology is the second generation of molecular marker technology, not as stable as DNA barcode technology
Moreover, although researchers continue to improve the application of DNA barcoding technology in plant species identification, there are still reports that any DNA barcode can be applied to the identification of all plants, so for specific plant species, experiments are still needed to find out Suitable gene sequences are used as DNA barcodes

Method used

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  • DNA identification method of Isodon serra(Maxim.)Kudo and Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara
  • DNA identification method of Isodon serra(Maxim.)Kudo and Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara
  • DNA identification method of Isodon serra(Maxim.)Kudo and Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Screening of DNA barcodes and establishment of a gene bank including rbcL gene sequence fragments

[0059] In this example, with regard to the screening of the DNA barcode genes of Xihuangcao, Xianwen Xiangchacai and Xianwen Xiangchacai (variety Xianhuaxiangchacai), the most suitable ones were selected from the 4 sequences of ITS2, matK, psbA-trnH, and rbcL. The DNA barcodes for the identification of Xihuangcao and Xianwenxiangchacai, that is, rbcL gene sequence fragments. In addition, this example established a gene bank including the rbcL gene sequence fragments of C. chinensis and C. chinensis.

[0060] 1 Instruments, materials and reagents

[0061] 1.1 Materials

[0062] At 7 sampling points (Guangdong, Fujian), a total of 41 samples of Xihuangcao, Xianwen Xiangchacai and Xianwen Xiangchacai (variety Xianhuaxiangchacai) were collected, and the details are shown in Table 1 below. The following barcode screening experiments use plant leaves as materials...

Embodiment 2

[0104] Example 2 Identification of the Plant Species of Unknown Creek Yellow Grass and Streakia japonica

[0105] In this example, the method provided by the present invention is used to identify the samples of Unknown Xihuangcao and Cercis chinensis plant species.

[0106] Select the suspected but unidentified, fresh or dried whole grass of Xihuangcao and Xianwen Xiangchacai, and divide them into two parts, one part is identified by traditional taxonomic methods. Another copy was identified using the method used in the present invention.

[0107] Sample preparation for the method used in the present invention: unless otherwise specified, the surface of the medicinal material is scrubbed with 75% ethanol aqueous solution and then dried, and 5 mg to 2 g of leaves are weighed for later use.

[0108] The steps described in Example 1 were used to extract genomic DNA from leaves of plant tissues, and the rbcL gene sequence fragment was amplified using SEQ ID NO.42 and SEQ ID NO....

Embodiment 3

[0115] Example 3 Identification of Species of Unknown River Huangcao and Xianwen Xiangchacai Decoction Pieces

[0116] In this example, the method provided by the present invention is used to identify the species samples of Unknown Xihuangcao and Xianwen Xiangchacai decoction pieces.

[0117] The suspected but unidentified decoction pieces of Xihuangcao and Xianwen Xiangchacai were selected and divided into two parts, one part was identified by traditional taxonomic methods. Another part is identified by the method used in the present invention, and the identification steps are the same as in Example 2.

[0118] After identification by this method, the conclusion obtained is consistent with that of the traditional method.

[0119] However, after comparison, the requirements of this method for samples are easier to obtain than those of traditional methods.

[0120] The sample required by the method is a part of the plant where plant DNA is easily obtained. Although the deco...

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Abstract

The invention provides a gene pool containing rbcL gene sequence fragments of labiate Isodon serra(Maxim.)Kudo and Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara. The invention also provides a method for identifying plant species of the labiate Isodon serra(Maxim.)Kudo and Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara. The method comprises the step of comparing a rbcL gene or its sequence fragment of a plant sample, plant species of which is to be identified, to the gene pool provided by the invention. The invention also provides an application of the rbcL gene or its sequence fragment in identifying the labiate Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara and the labiate Isodon serra(Maxim.)Kudo. It proves that the rbcL sequence is general, is easy to amplify and compare, and has good amplification and identification effects on the labiate Rabdosia lophanthoides (Buch.-Ham. ex D. Don) Hara var. graciliflora (Benth.) Hara and the labiate Isodon serra(Maxim.)Kudo.

Description

technical field [0001] The invention belongs to the field of identification of plant species, and in particular, the invention relates to a method for identification of species of xihuangcao and threaded fragrant tea vegetables. Background technique [0002] Xihuangcao, a traditional Chinese medicine, is a traditional medicine used by residents in Fujian, Guangdong. It can relieve dampness and reduce jaundice. It is used for the treatment of hepatitis, acute and chronic cholecystitis, and dysentery. It is also used as a herbal tea for daily health care. [0003] In the field of traditional Chinese medicine, the names of some Chinese medicinal materials are inconsistent with their origin, that is, the plant taxonomic name. This situation exists in the traditional Chinese medicine Xihuangcao. The traditional Chinese medicine Xihuangcao used in the folk tradition is the Lamiaceae plant line grain fragrant tea dish [Latin name Rabdosia lophanthoides (Buch.-Ham.ex D.Don) H.Hara o...

Claims

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Application Information

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IPC IPC(8): C40B40/08C12Q1/68
Inventor 张慧晔马新业李楚源邓乔华刘峰
Owner GUANGZHOU BAIYUSN HUTCHISON WHAMPOA CHINESE MEDICINE
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