Camellia polymorphic chloroplast genome microsatellite molecular marker primers and method for screening and discriminating related species

A technology of chloroplast genome and polymorphic primers, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve problems such as flooding of false seedlings, market influence on seedling production links, etc.

Active Publication Date: 2019-02-15
JIANGXI ACAD OF FORESTRY
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Problems solved by technology

Another object of the present invention is to solve the problem that Zhejiang red camellia fake seedlings flood the market and affect its seedling production link, and use some cpSSR polymorphic primers to screen Zhejiang red camellia and its close relatives

Method used

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  • Camellia polymorphic chloroplast genome microsatellite molecular marker primers and method for screening and discriminating related species
  • Camellia polymorphic chloroplast genome microsatellite molecular marker primers and method for screening and discriminating related species
  • Camellia polymorphic chloroplast genome microsatellite molecular marker primers and method for screening and discriminating related species

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Embodiment 1

[0035] 1. Screening of polymorphic primers for the cpSSR molecular marker of Camellia

[0036] 1. Mining and screening of chloroplast microsatellite molecular markers

[0037] (1) Acquisition and pretreatment of the cpDNA sequence of Zhejiang Red Camellia

[0038]Firstly, the chloroplast suspension was obtained through the Chloroplast DNA Isolation kit, and then the chloroplast DNA of Zhejiang Hongshan tea leaves was extracted with the DNeasyPlant Mini Kit to construct a sequencing library. After the sequencing data were screened to remove low-quality sequences, SOAPdenovo was used for skeleton assembly (the optimal Kmer was 31), and Newbler and MITObim were used to assist assembly, and finally GapCloser was used to fill holes, and finally the complete gene sequence of Zhejiang Red Camellia chlorophyll (MG431968 ).

[0039] (2) Screening of cpSSR polymorphism primers

[0040] From the GenBank public database (http: / / www.ncbi.nlm.nih.gov / ) download the Camellia genus Camelli...

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Abstract

The invention belongs to the technical field of forestry molecular biology and particularly relates to camellia polymorphic chloroplast genome microsatellite molecular marker primers and a method forscreening and discriminating related species. Based on the relative consistency of camellia species chloroplast DNA sequence microsatellite sites, cpSSR primers are designed aiming at the specific microsatellite sites with interspecies variation; DNA of different camellia species is execrated, mixed with the primers and amplified and then is subjected to electrophoresis detection; finally, the 16pairs of camellia polymorphic chloroplast genome microsatellite molecular marker primers are obtained. The method for discriminating the related species comprises the following steps: extracting bothDNA of Zhejiang red camellia and DNA of related species of the Zhejiang red camellia, performing capillary electrophoresis separation by adopting the polymorphic primers, and separating and discriminating the related species according to PCR product fragments. The primer screening method provided by the invention is quick, efficient and widely applicable to other plant species with chloroplast genome sequence information; an obtained polymorphic cpSSR marker can be used for the systematic classification of camellia plants, related species discrimination, genetic diversity analysis and other studies.

Description

technical field [0001] The invention relates to the technical field of forestry molecular biology, in particular to a polymorphic primer of a chloroplast microsatellite molecular marker of the genus Camellia, a screening method thereof, and a method for discriminating close relatives. Background technique [0002] Camellia (Camellia L.) is the largest genus in the family Theaceae, and many species in the genus have important economic value. For example, tea (C. sinensis) in the tea group is one of the three major beverages in the world; The seeds of the species have high oil content and are edible, the most famous of which is Camellia oleifera (C. oleifera), which is called the oriental olive tree; in addition, many species of the red camellia group have great ornamental value, such as the internationally famous camellia (C.japonica), as well as ornamental and oil species such as Zhejiang red camellia (C.chekiangoleosa). The relationship between the species of Camellia is c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 温强殷鑫刘丽婷李田王建文杨军朱恒叶金山徐立安
Owner JIANGXI ACAD OF FORESTRY
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