Molecule tag in use for identifying strain of Porphyra haitanensis and application

A technology of molecular markers and identification of laver, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of difficulty in satisfying the cultivation and production of laver, time-consuming and labor-intensive, and quality decline, so as to shorten the breeding cycle and use Convenience and low detection cost

Inactive Publication Date: 2005-08-31
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, there has been a common problem of quality decline in the algae laver cultivation industry. Therefore, it has become very urgent to use molecular biology methods combined with traditional breeding methods to cultivate new strains of high-quality algae laver.
At present, the breeding of fine strains of algae laver mainly adopts field selection, physi

Method used

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  • Molecule tag in use for identifying strain of Porphyra haitanensis and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Identification of the acquisition of AFLP markers for the excellent strains of Porphyra alba

[0025] 1. Materials: See Table 1 for the names and sources of 16 laver samples used in the experiment (stored in the shade at -20°C).

[0026] breed name

source

growing conditions

breed name

source

growing conditions

GL1

Fujian Pingtan

artificial breeding

XSD

Fuqing City, Fujian Province

wild breeding

breed

LSD

Fujian Pingtan

the wild

NH

Fujian Pingtan

the wild

BBD

Fujian Pingtan

the wild

DXY

Fujian Pingtan

wild breeding

breed

JJS

Fujian Pingtan

the wild

LH

Longhai City, Fujian Province

artificial breeding

SYS5

Xiamen University

Radiation Mutation Breeding

LPZ

Fujian Pingtan

artificial breeding

NH1 ...

Embodiment 2

[0051] Example 2. Acquisition of excellent strain-specific SCAR markers

[0052] 1. Materials: Laver samples are the same as in Example 1, Escherichia coli DH5α, pMD18-T Vector

[0053] 2. Method:

[0054] 1. Recovery of specific AFLP fragments: According to the obtained AFLP polypropylene gel electrophoresis pattern, look for the specific AFLP marker band of the excellent strain, and cut off the gel containing the band with a sharp blade. The recovered gel pieces were dissolved in 400 μL of high-salt solution (20% ethanol, 1mol / L LiCL and 10mmol / L Tris-HCL, Ph7.5), placed at room temperature for 24 hours, incubated at 65°C for 2 hours, and the DNA was precipitated with absolute ethanol and Dissolve in 20 μL TE.

[0055] 2. PCR re-amplification of specific AFLP fragments: Take 5 μL of recovered DNA as a template, use the same primers as in selective amplification, and amplify for 35 cycles at 94°C for 30S; 56°C for 30S; 72°C for 60S according to the following procedure.

[...

Embodiment 3

[0075] Example 3. PCR primer sequences obtained by SCAR markers

[0076] 1. Method

[0077] Using the SCAR marker sequence of ACA-CAC (203) as a primer design template, primers were designed by Primer 5.0 primer design software.

[0078] 2. Primer Design Principles

[0079] 1. The GC content in the two primers should be as similar as possible;

[0080] 2. Avoid obvious secondary structure inside the primer;

[0081] 3. Avoid complementary sequences between the two primers to avoid the formation of "primer dimers";

[0082] 4. The pairing of primers and target sequence fragments must be precise and strict, especially at the 3' end;

[0083] 5. The specific primer should have a certain annealing temperature (not lower than 50°C) and a certain length (not less than 18 bp).

[0084] 3. Results

[0085] Specific primers were designed according to the SCAR marker sequence, and its sequence is:

[0086] Marker1: 5’-AGT AAA ATG GGG AAC AGC-3’

[0087] Marker2: 5'-ATC GCA GAA T...

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Abstract

A molecular marker for discriminating the excellent variety of ampullaceous laver is disclosed. Its application includes using AFLP marker technique to analyze the excellent varieties of ampullaceous laver, finding out the AFLP marker specific to excellent variety, converting it to SCAR marker, designing specific primer Marker and Marker 2, preparing reagent kit, PCR amplifying to individual, and discriminating. It can be also be used for marker said seed culture.

Description

technical field [0001] The invention relates to a characteristic sequence amplification (Sequence Characterized Amplified Region, SCAR) marking technology and its application in germplasm purity identification, strain identification and molecular marker assisted breeding. Background technique [0002] Porphyra haitanensis belongs to Rhodophyta (Rhodophyta), Bangiacea (Bangiacea) Porphyra, is one of the main varieties of artificial laver cultivation, has high nutritional value and economic value, and its economic benefits are relatively high. it is good. However, in recent years, there is a common problem of quality decline in the algae laver cultivation industry. Therefore, it is very urgent to use molecular biology methods combined with traditional breeding methods to cultivate new strains of fine almanac laver. At present, the breeding of fine strains of algae laver mainly adopts field selection, physical mutagenesis or chemical mutagenesis, and somatic cell engineering s...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 左正宏陈奕欣李博文王重刚陈骁赵扬陈昌生
Owner XIAMEN UNIV
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