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Method for identifying opium poppy by using chloroplast DNA bar code

An identification method and barcode technology, applied in the field of molecular identification, can solve the problems of low accuracy, unsafe, poor reliability, etc., and achieve the effect of fast detection process and high accuracy of results.

Inactive Publication Date: 2018-08-14
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing identification methods have the problems of spending a lot of time, wasting manpower and material resources, and also have poor reliability, low accuracy, and even unsafe conditions.

Method used

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  • Method for identifying opium poppy by using chloroplast DNA bar code
  • Method for identifying opium poppy by using chloroplast DNA bar code
  • Method for identifying opium poppy by using chloroplast DNA bar code

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The present invention increases the number of individuals as much as possible by widely sampling the plants of the genus Papaver distributed in China, constructs a database of chloroplast DNA barcodes of the plants of the genus, finds the unique information sites of the DNA barcodes of poppy (opium) by comparison, and further obtains its unique DNA barcode.

[0025] The poppy DNA barcode of the present invention is amplified from four fragments matK, psbA-trnH, rbcL and trnL-trnF, and the total length is 2645bp. In this sequence, the specific recognition sites are: 105bp is A, 121bp is A, 333bp is A, 748bp is T, 1539bp is G, 1751bp is A, 2392bp is G, 2604bp for A. What the amplification primer of the present invention adopts is common DNA bar code universal primer, there are 8 specific recognition sites in the sequence of 2645bp that amplifies, can identify opium poppy (opium) according to the base type of above-mentioned specific site .

[0026] The poppy DNA barcod...

Embodiment 2

[0047] 1. Collection and protection of specimens of Poppy plants:

[0048] Consult the literature and the collection information of domestic herbaria, formulate a sampling plan according to the sampling requirements of DNA barcodes, collect individuals from different sources in the distribution area of ​​each species as much as possible, and dry the leaves of all individuals with silica gel. There are 7 species of Papaver, and the total number of individuals is the leaf material of 25 individuals (including variants). The sequence information of some samples is downloaded from the NCBI gene bank (total 165 sequences). Among them, the samples of opium poppy (opium) came from wild sporadic growth in the wild in Ili, Xinjiang, and 3 leaf samples were collected from each location. See Tables 1 and 2 for the list of materials and sources of samples.

[0049] Table 1 Specimens and sources of Papaver

[0050]

[0051] Table 2 Primers and Genebank numbers of Poppy plants

[0052...

Embodiment 3

[0092] With the flowerless and fruitless poppy and other poppy leaves collected in medicinal planting areas and in the field as materials, the method in Example 1 was used for DNA extraction, PCR amplification and sequencing, and the identification results showed that in the known poppy plants The 105th bp of the obtained DNA barcode sequence was A, the 121st bp was A, the 333rd bp was A, the 748th bp was T, the 1539th bp was G, the 1751st bp was A, the 2392th bp was G, and the 2604th bp was A. However, the above-mentioned specific sites were not found in other unknown plants of the genus Papaver, so the samples of Papaver could be identified.

[0093] The sequence list of the present invention is the trnH, matK, rbcL, trnL sequence of a poppy (Papaver somnferium) individual of the present invention, and there are 7 bases missing in comparison with other species.

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Abstract

The invention provides a method for identifying papaveraceae papaver opium poppy (opium) by using chloroplast gene DNA bar code. According to the method, the distribution region of papaver plants is widely surveyed; samples in a plurality of places are collected; a database of a chloroplast sequence of the plant in the papaver is built; through comparison, special information sites of the opium poppy are discovered; in the sequence with the total length of 2645bP, the site at105bp is A; the site at 121bp is A; the site at 333bp is A; the site at 748bp is T; the site at 1539bp is G; the site at1751bp is A; the site at 2392bp is G; the site at 2604bp is A. By using the differences between the sites and other species, the opium poppy can be fast and accurately identified.

Description

technical field [0001] The invention belongs to the technical field of molecular identification, and in particular relates to an identification method of a chloroplast DNA molecular barcode, which performs DNA barcode sequencing and molecular identification on poppy (Papaver somniferum). Background technique [0002] Poppy (Papaver somniferum L.), also known as opium and opium, is known as the world's three major drug plants along with marijuana and coca. It has always been a taboo, and its cultivation and use are strictly controlled. It is native to southern Europe and is a plant of the genus Papaver (Papaver L.) in the family Papaveraceae. There are about 700 species in the family Papaveraceae, and about 100 species in the genus Papaver. It is mainly distributed in the temperate zone from Europe to Asia, and a few species are produced in America, Oceania and Africa. There are 7 species in my country, namely: poppy (Papaversomniferum), poppy (P.rhoeas), black ring poppy (...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6895C12Q2600/156C12Q2531/113
Inventor 杨俊波杨志云伊廷双李洪涛林春燕曾春霞杨继雄
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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