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Molecular genetic linkage mapping method of cotton

A genetic linkage, cotton technology, applied in the field of cotton molecular genetic linkage mapping, can solve the problems of high DNA requirements, time-consuming development of SSR, limited SSR, etc., to achieve high marker yield, overcome uneven marker distribution, and ensure accuracy. Effect

Inactive Publication Date: 2004-11-03
HUAZHONG AGRI UNIV
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Problems solved by technology

Restriction fragment length polymorphism (RFLP) markers have the advantages of co-dominance, complete information, good repeatability and stability, etc., but the experimental operation process of RFLP technology is complicated, it is not easy to realize automation, and the requirement for DNA is high, and the dosage Large, expensive to analyze large groups
Randomly amplified polymorphic DNA (RAPD) markers require less DNA, have low requirements for DNA, and are easy to operate, but RAPD markers are easily affected by experimental conditions, resulting in poor reproducibility and low yield
Simple sequence repeats (SSRs) are a good marker, but cotton has limited SSRs, and developing SSRs is time-consuming and expensive
The linkage map density of molecular markers in cotton is not enough, and markers must be added to meet the needs of the application. The existing markers have limited ability to increase the map density, and new markers must be applied

Method used

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  • Molecular genetic linkage mapping method of cotton
  • Molecular genetic linkage mapping method of cotton
  • Molecular genetic linkage mapping method of cotton

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Embodiment 1

[0031] 1. Using the sea-island cotton line Pima90 as the male parent and the upland cotton line Han 208 as the female parent, a hybrid combination was prepared, and the obtained F 2 A single plant is used as the starting material for genetic linkage mapping, and the material is planted in the field;

[0032] 2, get described parent and F from described field material 2 The total DNA was extracted from the tender green leaves of a single group plant;

[0033] 3. Using SRAP markers to construct a linkage map of cotton molecular markers, a 39 linkage groups containing 237 loci were obtained, with a total length of 3030.7cM, covering 65.4% of the entire genome, and an average distance between markers of 12.79cM. This is the first molecular genetic linkage map of cotton constructed by SRAP. Specific steps are as follows:

[0034] 3.1 SRAP marker analysis:

[0035] SRAP analysis: PCR primers except Li and Quiros (Li G, Quiros C F. Sequence-related amplified polymorphisim (SRAP),...

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Abstract

The present invention is linked inheritance mapping method utilizing SRAP molecular mark for cotton. The mapping population is hybridization between sea island cotton and upland cotton to generate F2 single plant. Applying SRAP molecular mark and PCR proliferation for detecting population polymorphism in inheritance mapping obtains 285 polymorphic bands in high yield. The 285 marks are used in constructing MAPMAKER linkage group, and 237 marks enter 39 linkage groups, each of which has 2-13 marks. All the linkage groups have the total length of 3030.7 cM, covers 65.4 % of the total cotton genome, and the average mark interval is 12.79 cM. The marks are distributed in the linkage groups relatively homogeneously. The specific molecule marking steps and electrophoresis parameters are also proposed.

Description

technical field [0001] The invention belongs to a method for evaluating cotton linkage genetics, in particular to a method for cotton molecular genetic linkage mapping, in particular to the invention of a method for cotton molecular genetic linkage mapping using sequence-related amplified polymorphism (SRAP) molecular markers . Background technique [0002] Molecular genetic linkage maps have been constructed in a variety of plants. The construction of cotton molecular marker linkage map began in 1994. Reinish applied restriction fragment length polymorphism (RFLP) markers to construct a map containing 705 polymorphic sites distributed into 41 linkage groups of different sizes. Total length 4675cM (Reinisch A J, Dong J M, Brubaker C L, StellyD M, Wendel J F, Paterson A H.A detailed RFLP map of cotton Gossypium hirsutum×Gossypium barbadense: chromosome organization and evolution in a disomic polyploid genome. Genetics, 1994, 138: 829 ). Sha...

Claims

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Application Information

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IPC IPC(8): A01H1/02C12N15/10C12Q1/00
Inventor 张献龙林忠旭聂以春贺道华
Owner HUAZHONG AGRI UNIV
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