Wheat BSR-Seq gene locating method

Abstract

The invention discloses a wheat BSR-Seq gene locating method. The wheat BSR-Seq gene locating method comprises the following steps of: constructing and sequencing mixing pools, performing quality variation mining, screening a transcript tightly linked to a target gene, developing and locating a molecular marker, etc. The next-generation transcriptome sequencing technology (transcriptome sequencing, RNA-Seq) and a mixing pool technology (Bulked Segregant Analysis BSA) are combined; a wheat sequencing draft sequence is used as a reference sequence at first; then, a lot of high-quality SNP heritable variations on the transcript is mined at high throughput by adopting the next-generation sequencing technology; the allele frequency is precisely calculated in combination with the mixing pool technology, so that the transcript tightly linked to the target gene possibly can be rapidly screened out; false positive is precisely checked and controlled through Fish; the method is independent of a reference genomic sequence, low in cost, rapid and high in precision; the wheat gene locating efficiency and precision are increased; the wheat polymorphic molecular marker developing cost is reduced; therefore, the wheat gene fine locating working time is reduced to several months from several years; the locating precision is reduced to a fraction or 0 cM from several cM; and the fine locating cost is reduced to several thousands from several ten thousands.

Description

technical field [0001] The invention relates to a genetic breeding technology, in particular to a wheat BSR-Seq (Bulked SegregantRNA-Seq, mixed pool transcriptome sequencing) gene positioning method. Background technique [0002] Wheat is one of the main ration crops for human beings. Under the background of decreasing arable land and increasing demand, its production is critical to food security. Breeding high-yielding, high-quality, stress-resistant wheat varieties and genetically improving important agronomic traits of wheat are important methods for stabilizing and increasing wheat yield. Improving the efficiency of traditional wheat breeding methods, continuous innovation and application of new breeding techniques are the only way to go. At present, molecular marker technology, marker-assisted selection breeding and genome selection are getting more and more attention. The use of closely linked molecular markers to identify and screen chromosome segments or genes relat...

AI Technical Summary

Problems solved by technology

However, the high-quality SNP markers obtained by this technology are limited, and the high-copy characteristics of wheat genes are prone to errors, which requires high sequencing costs for each individual, so its application is limited

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