Method for building cotton fiber transcription genetic linkage map by EST-SSR sign

Abstract

The invention belongs to the technical field of cotton molecule breeding, particularly relating to a method for building a cotton fiber transcription genetic linkage map by EST-SSR signs. The method comprises the following steps: taking gossypium barbadense Pima 3-79 as a male parent and Gossypium hirsutum Emian 22 as a female parent, and hybridizing to obtain F1; planting F1, and causing F1 to be inbreeded to obtain F2; taking the F2 single plant as a starting material for the fiber transcription map to be plotted; extracting the RNA of the fiber of the field F2 single plant in 5 days after the single plant blooms, and carrying out reverse transcription of the RNA into cDNA to be served as a plotting group of the transcription map; utilizing the reported EST-SSR primer and the primer shown by a self-designed sequence table SEQ ID NO:1-90 to carry out the plotting group analysis to the F2 single plant by a denatured polyacrylamide gel electrophoretic analysis; and finally, analyzing data by MAPMAKER / EXP.3.0 mapping software, and manufacturing and obtaining the transcription map. Compared with the existing method, the method of the invention is simple and practical, has accurate QTL positioning and is convenient for cloning genes relevant to cotton fiber development.

Description

technical field [0001] The invention belongs to the technical field of cotton molecular breeding. Specifically, it relates to a method for constructing a genetic linkage map of cotton fiber transcription using EST-SSR 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. Since Reinish applied restriction fragment length polymorphism (RFLP) markers in 1994 to construct a cotton genetic linkage map containing 705 polymorphic sites and a total length of 4675cM, many researchers (Rong, J., C.Abbey , et al, 2004; Nguyen, T., M.Giband, et al, 2004; He, D., Z.Lin, et al, 2007; Guo, W., C.Cai, et al, 2007; Zhang, Y., Z.Lin, et al, 2008) also constructed multiple cotton genetic linkage maps using different marker methods. [0003] However, the previous markers were all carried out at the DNA level, which has its limitations: firstly, the previous ge...

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Problems solved by technology

[0003] However, the previous markers were all carried out at the DNA level, which has its limitations: firstly, the previous genetic linkage maps used randomly distributed markers to locate the QTL of fiber traits, and the results obtained by different researchers were almost inconsistent; secondly, biological The information content of RNA is far greater than that of DNA, and it is impossible to study the "compressed" part of genetic information at the DNA level

[0005] However, using the cDNA-AFLP strategy to construct transcription maps has many disadvantages: first, the AFLP operation process is cumbersome, requiring enzyme digestion and two amplifications; second, the quality of cDNA is very high, and some crops (such as cotton) The RNA extraction is complicated and the quality is not high, and the double-stranded cDNA obtained by reverse transcription is difficult to reach the quality and concentration of enzyme digestion; the third is the high cost, which is not suitable for population analysis and mapping

[0009] There is no report on the method of using EST-SSR markers to construct transcriptional genetic linkage maps of cotton fibers

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