Development And Application Of High-throughput Molecular Detection Methods On The TaCO9-1A In Wheat

Wheat is one of the world’s most important food in daily life. However, with the global warming climate, the production of wheat is facing great challenges. Jarovization and photoperiodic responses are important characteristics of wheat, they not only directly affect the use of wheat varieties、planting regionalization、introduction and cultivation techniques, but also indirectly affect some important agronomic characters, such as yield、disease resistance. So, it is of great significance for breeding by excavating the new photoperiod genes, studying their function and applying high-througput detection method with the meocular maker.Our lab have got the wheat photoperiod homologous gene HvCO9-TaCO9 by using of the comparative genomics-homology-based cloning technology. A preliminary analysis found it is springness varieties in Ningchun4, analysis revealed that in the second exon 321 sites, there are 6 bp insertion in TaCO9 genes. Then, we used Chinese spring material with deficiency-four segments, located its deficiency in spring wheat chromosomes 1A, named TaCO9-1A.Based on this, we developed specific molecular markers. Polypropylene acyl ammonia gel electrophoresis(PAGE) and high resolution melting curve(HRM) techniques are used to explore TaCO9-1A genetic polymorphisms and their correlation with wheat growth habit. We aims to providing a theory basis in widely adaptive wheat cultivars, the distribution of wheat varieties、regional test. The conclusions are as followed:1. We confirmed that there exactly exist 6 bp differences.Through the comparison of strong winter wheat ZhongMai895 and spring wheat NingChun4 TaCO9-1A gene sequences, we designed the specific molecular marker, named TSW.2. Polymorphism test were conducted by using the designed markers and 223 wheat varieties(natural). Polypropylene acyl ammonia gel electrophoresis(PAGE) results showed that 99 materials among the natural popution have the same belt type(TS) with NingChun4, 133 materials have the same belt type(TW) with Zhongmai895.3. We used high resolhongution dissolution curve(HRM) technology for genotyping in natural population wheat materials, and they were consistent with the results of polyacrylamide gel electrophoresis.So, HRM analysis can use of the high-throughput molecular detection.4. Using the molecular markers TSW,genetic polymorphism analysis of TaCO9-1A gene in F2 group is conducted, PAGE gel electrophoresis showed three bands, and the separate ratiao are 1:2:1. This proves, use the tag can be done group genotype selection in the separation of breeding.5. We investigated the heading stage and the flowering period of wheat field material. Correlation analysis found that there is a significant correlation in spring and winter(P < 0.01).This conclusion can assist us development and application of the tag and highthroughput molecular detection method have vital significance for wide eligibility breeding of new varieties of wheat, winter and spring testing and breeds division.