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RAPD special primers and method for identifying genus and species of common melastoma herb

A technology of peony and peony leaves, applied in the molecular field

Inactive Publication Date: 2016-10-12
FUJIAN INST OF TROPICAL CROPS
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the taxonomic research on the genus of P. peony mainly focuses on macroscopic morphology, cytogenetics, palynology, cytology, and phenotypic diversity. The use of molecular markers to study the heritability of P. peony germplasm resources is relatively incomplete. few

Method used

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  • RAPD special primers and method for identifying genus and species of common melastoma herb
  • RAPD special primers and method for identifying genus and species of common melastoma herb
  • RAPD special primers and method for identifying genus and species of common melastoma herb

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Embodiment 1

[0023] See Table 1. The test materials in this study are 44 wild germplasm resources of the genus Peony, which come from five provinces: Fujian, Guangdong, Guangxi, Hainan, and Yunnan. Collected fresh leaves were frozen in liquid nitrogen and stored at -80°C.

[0024] Table 1 Germplasm resources of 44 tested species of Mesophyllum

[0025] Table 1 44 individuals of Melastoma

[0026]

[0027]

Embodiment 2

[0028] Example 2 DNA Extraction

[0029] The improved CTAB method determined in the previous experiment was used to extract the genomic DNA of the wild peony. The main steps are as follows:

[0030] (1) Take about 1g of fresh young leaves, put them into a pre-cooled mortar, quickly grind them into powder in liquid nitrogen, transfer them to a pre-cooled 10mL centrifuge tube, add 4mL of CTAB extract preheated to 65°C At the same time, 80 μL of β-mercaptoethanol was added, slowly mixed up and down, and placed in a water bath at 65° C. for 60 minutes (during this period, the mixture was mixed upside down once every 10 minutes).

[0031] (2) Take out the centrifuge tube and cool it down to room temperature, add an equal volume of chloroform / isoamyl alcohol (24:1), mix well, shake slowly on a horizontal shaker for 30 minutes, and after standing still, centrifuge at 4°C and 7500g for 5 minutes to recover upper (water) phase.

[0032] (3) Add 1 / 10 volume of CTAB / NaCl solution (65° ...

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Abstract

The invention discloses RAPD special primers and a method for identifying genus and species of common melastoma herb, wherein molecular markers include the following two primers: 1) S19 having the sequence of ACCCCCGAAG, and 2) S43 having the sequence of GTCGCCGTCA. The method comprises the steps of firstly, establishing an RAPD fingerprint spectrum of the common melastoma herb, then carrying out amplified product gel electrophoresis and comparing with the fingerprint spectrum. The method has the following advantages: 30 common melastoma herb germplasm resources can be identified by using the primer S19, 29 common melastoma herb germplasm resources can be identified by using the primer S43, and with adopting of the combination of the primers S19 and S43, 44 common melastoma herb germplasm resources for test can be identified.

Description

technical field [0001] The invention relates to the field of molecular technology, in particular to RAPD special primers and a method for identifying wild peony species. Background technique [0002] There are about 240 genera and more than 3000 species in the Melastomaceae plant in the world. The genus Melastoma belongs to the family Melastomaceae, and there are about 100 species. The distribution center is in Southeast Asia. There are 9 species and 1 variety in my country, and the main distribution areas are in various provinces and regions south of the Yangtze River. The flower colors of wild peonies are white, pink, and purple. They have a long flowering period and a large amount of flowers. The plant types are distributed from ground cover, shrubs to small trees, and have good ornamental properties. In addition, the medicinal properties of this genus are The value has also been studied in depth recently, and its development prospects are broad. At present, the vast ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 林艺华陈振东苏金强郑涛林秀香
Owner FUJIAN INST OF TROPICAL CROPS