Method for identifying lines of lonicera japonica thunb (Damaohua and Jasminum duclouxii) based on PCR-RFLP

A technology of honeysuckle and chicken feet, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., and can solve the problems of glandular hair loss and indistinguishability.

Active Publication Date: 2015-02-04
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

However, the morphology of Damaohua and Gallina japonica are similar at the seedling stage, while the commercial honeysuckle in the market has changed in shape after processing, transportation and storage, and the glandular hairs fall off, making it difficult to distinguish

Method used

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  • Method for identifying lines of lonicera japonica thunb (Damaohua and Jasminum duclouxii) based on PCR-RFLP
  • Method for identifying lines of lonicera japonica thunb (Damaohua and Jasminum duclouxii) based on PCR-RFLP
  • Method for identifying lines of lonicera japonica thunb (Damaohua and Jasminum duclouxii) based on PCR-RFLP

Examples

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Effect test

Embodiment 1

[0051] Example 1. Preparation of kits for identification of honeysuckle strains Damaohua and Gallinarum japonica and establishment of identification methods

[0052] 1. Composition of kits for identification of honeysuckle strains Damaohua and Chickenpaw

[0053] The present invention is used to identify and identify the honeysuckle strains Damaohua and Chizuna. The kit is designed based on PCR-RFLP technology and contains a pair of primers composed of two single-stranded DNA molecules shown in sequence 1 and sequence 2 in the sequence table , and the restriction enzyme MfeI. The design process is roughly as follows:

[0054] The inventors of the present invention analyzed the COi gene sequences related to the development of glandular hairs of the honeysuckle strains "Da Maohua" and "Chicken's Claw", obtained the SNP sites of differences between "Da Maohua" and "Chicken's Claw", and found that there is a SNP The existence of the site (recorded as SNP site A) caused the recog...

Embodiment 2

[0076] Example 2, Effective verification of the method for identifying honeysuckle strains Damaohua and Chickenpaw based on PCR-RFLP

[0077] Tested honeysuckle: 75 known lines of honeysuckle from different origins shown in Table 1.

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Abstract

The invention discloses a method for identifying lines of lonicera japonica thunb (Damaohua and Jasminum duclouxii) based on restrictive fragment length polymorphism polymerase chain reaction (PCR-FRLP). The invention provides a kit for identifying lines of lonicera japonica thumb (Damaohua and Jasminum duclouxii) which comprises a primer pair composed of two single-stranded DNA molecules as shown in sequence 1 and sequence 2 in a sequence table as well as restriction enzyme MfeI or isoschizomer thereof. The method disclosed by the invention, by analyzing COi gene sequences related to development of glandular hairs of lines of lonicera japonica thumb (Damaohua and Jasminum duclouxii), can obtain a difference SNP site of Damaohua and Jasminum duclouxii; identification primers and corresponding restriction enzyme are designed; and accurate identification of two lines of the lonicera japonica thumb is realized through the PCR-FRLP technology.

Description

technical field [0001] The invention belongs to the field of identification of plant varieties, and relates to a PCR-RFLP-based method for identifying honeysuckle strains Damaohua and Chickenpaw, in particular to a PCR-RFLP-based kit for identifying honeysuckle strains Damaohua and Chickenpaw and its identification method. Background technique [0002] The base plant of the medicinal material honeysuckle is Lonicera japonica Thunb., and its morphology has undergone obvious variation and differentiation during the long-term cultivation process, forming many farm cultivars or strains. The investigation of the traditional honeysuckle production areas in Shandong and Henan found that the honeysuckle cultivars "Damaohua" and "Chicken's Foot" are the mainstream commodities of honeysuckle, and there are certain differences in their original plant morphology, medicinal properties, and chemical components. "Da Maohua" is named for its leaves and buds densely covered with hairs. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2521/301C12Q2525/131
Inventor 黄璐琦袁媛蒋超胡添源
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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