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EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application

A technology of upland cotton and markers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems that have not been reported in the development, and achieve rich polymorphism, stable amplification, and uniform distribution Effect

Active Publication Date: 2015-06-24
SHANDONG COTTON RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the EST-SSR developed in cotton is mainly related to fiber development genes, and the development of SSR markers for genes related to cotton early morphogenesis has not been reported yet.

Method used

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  • EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application
  • EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application
  • EST-SSR marker primers developed on basis of upland cotton transcriptome sequence and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0042] Application of EST-SSR in Coker312 Transcriptome of Upland Cotton in Interspecific and Intraspecific Polymorphism Analysis of Cotton

[0043] 1. Extraction of cotton genomic DNA

[0044] (1) Material

[0045] A total of 13 materials from 5 cotton species were selected from the cotton germplasm materials (see Table 1) for the evaluation of newly developed primers, including amplification effect and polymorphism analysis.

[0046] Table 1 is used for the material information of primer polymorphism analysis in the present invention

[0047]

[0048] (2) Genomic DNA extraction by CTAB method

[0049] Cotton DNA was extracted by the improved CTAB method, and the specific steps were as follows:

[0050] 1) Take 0.2 g of fresh leaves of the cotton material to be tested, wash them, dry them, shred them, put them into a 2 mL centrifuge tube A, add liquid nitrogen, and grind them with a Retsch (MM400) tissue grinder.

[0051] 2) Add 700 μL of extraction buffer to the centr...

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Abstract

The invention relates to EST-SSR marker primers developed on the basis of an upland cotton transcriptome sequence and an application. EST-SSR markers are respectively SCRC056, SCRC371, SCRC878 and SCRC1022. The invention also relates to the application of the EST-SSR marker primers in genetic diversity analysis and high-density genetic map construction. The EST-SSR primers developed on the basis of the upland cotton stem tip transcriptome sequence, disclosed by the invention, have the advantages of stable amplification, clear electrophoretic bands, rich polymorphism and the like, so that the EST-SSR primers can be effectively used in the research fields of genetic diversity analysis of cotton germplasm resources, construction of high-density maps, varietal purity and authenticity identification, molecular marker-assisted breeding and the like.

Description

technical field [0001] The invention relates to EST-SSR marker primers developed based on upland cotton transcriptome sequences and applications thereof, in particular to EST-SSR primers developed based on upland cotton Coker312 shoot tip transcriptome sequences and applications thereof, belonging to the technical field of molecular markers. Background technique [0002] Cotton is one of the important economic crops and the most important textile fiber crop. There are 4 cultivars, including 2 diploid cotton species, namely grass cotton (Gossypium. herbaceum) and Asian cotton (G. arboreum) and 2 tetraploid cotton species, namely upland cotton (G. hirsutum) and Sea Island cotton (G. barbadense). Due to its high yield and adaptability, upland cotton has become an important variety among cultivated species. However, the diversity level of upland cotton varieties currently bred is low, and the genetic basis is narrow, which makes it difficult to distinguish and identify in prod...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 张军高阳王芙蓉刘国栋张传云张景霞周娟
Owner SHANDONG COTTON RES CENT