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Tobacco (Tobacco) as an important economic crops, natural conditions and artificial selection training, and constantly introduced from abroad, China has formed a colorful variety of resources. Since 1986, the tobacco varieties-trial (share) since the system is fixed until 2010, China National Tobacco Variety Approval Committee approval (or approval) of 91 tobacco varieties, the promotion and application of these improved varieties of tobacco production has played a positive role in promoting. In this study, based on SSR fluorescent labeling and capillary electrophoresis detection technology, the establishment of a set of high-throughput tobacco varieties SSR markers detection system, and to establish the technical system of the main trial (recognized) as tobacco germplasm genetic diversity and phylogenetic relationship provide the scientific basis for tobacco breeding parent selection and use of favorable genes. The main findings are as follows: (1) the experimental based on SSR fluorescent labeling and capillary electrophoresis detection technology, by adjusting the amount of the PCR reaction components and cycling conditions, and ultimately established a set of suitable the automatic genetic analysis system of the Beckman CEQ8000 PCR amplification and detection the system. The capillary electrophoresis fluorescence detection technology and polyacrylamide gel electrophoresis technique for the detection result of the comparison shows that the fluorescence detection method to overcome the lack of silver staining, and has the advantages of simple, reliable, and high-throughput. Microsatellite fluorescent labeling of low-cost, high-throughput multiplex PCR system to establish some of the technical issues were discussed. Select 36 pairs of SSR primers 71 trial (recognized) the given the tobacco germplasm SSR amplification, produced a total of 2437 DNA bands were polymorphic 205; average each primer pair could amplify 2 to 15 , and the average polymorphism detection rate of 8.4%. Show that the efficiency of the SSR tobacco germplasm genetic diversity analysis. 3 based on Dice genetic similarity coefficients UPGMA cluster analysis, the results show that the clustering threshold of 0.35, flue-cured and burley into classes. , Tobacco groups can be further divided into 10 sub-categories, the Burley groups is divided into three sub-categories. In addition, tobacco groups, 13 of flue-cured tobacco germplasm from abroad are distributed in the domestic flue-cured tobacco breeding groups, the same place breeding tobacco varieties due to their location closer to the higher genetic similarity gathered in the same group or sub-category. 4 63 flue-cured tobacco germplasm and burley tobacco germplasm genetic diversity, to provide a scientific basis for tobacco breeding parent selection and use of favorable genes.
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