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Blue fruit honeysuckle (Lonicera Caerulea L.) is a the Caprifoliaceae (caprifoliaceae), honeysuckle (Lonicera L.), a perennial deciduous shrub with good environmental adaptability, mainly distributed in Russia, Japan, North Korea, and North America, China's large Mountains, Changbai Mountain also contains a wealth of wildlife resources; blue-purple fruit, its processing into jams, drinks, canned food, wine and other pollution-free food, both contain anthocyanins, minerals and vitamins and other biologically active substances, and has good medicinal value, and loved by people around the world, is one of the third generation of fruit trees and blueberries comparable market prospects. The experiment in 2009 to 2010 in the Northeast Agricultural University as test materials to the the germplasm nursery of the first Blue Lonicera 58 Blue Lonicera resources, established a blue fruit of honeysuckle leaves genome DNA. good extraction method, discussed the factors amplification results and reaction system optimized SRAP-PCR reaction system, the analysis of the 58 Blue Lonicera genetic diversity, understand blue Lonicera Home Genetic diversity and the protection and exploitation of the resources to provide theoretical basis. The main results are as follows: the separation and extraction of DNA is the basis of the analysis of genome diversity, the basic techniques of molecular biology research. Different plant material because of its tissue cells contained in the types of metabolites of this life, different content, the extraction method is also different. The blue fruit honeysuckle leaves are rich in polysaccharides, polyphenols and other secondary metabolites, these substances will affect the quality of the leaf genomic DNA extracted, the experiment is carried on the conventional CTAB and improved CTAB method compares the results show that the improved CTAB method of DNA strips and bright, no smearing, suitable Blue of Lonicera genome DNA extraction. Improved CTAB method with PVP powder in liquid nitrogen grinding process to prevent the oxidation of phenols with the SET nuclear separation buffer prewash samples, separated from the nucleus and secondary metabolites such as polysaccharides and polyphenols extract better OD 260 / OD 280 maintained at between 1.7 to 1.9, to meet the requirements of the SRAP amplification. 2, PCR reaction system, the factors used in an amount of each of the reaction results of the PCR reaction have a great influence of the experiment on the DNA tags in the reaction system polymerase, template DNA, dNTPs, the of Mg 2 and primers for single factor experiments, blue Lonicera SRAP-PCR reaction system (20 μl): 2.0μl 10 × PCR buffer, 1.0U Tag DNA polymerase, 20ng template DNA, 0.20 mmol / l dNTPs, 2.0mmol/lMg < sup> 2 , 0.3μmol / l primers. Reaction program was: 94 ° C denaturation 5min; denaturation at 94 ° C for 1 min, 35 ° C renaturation 1min, 72 ° C extension 90s, 5 cycle; denaturation at 94 ° C for 1 min, 50 ° C renaturation 1min, 72 ° C extension 90s, 35 cycle; 72 ℃ for 8min. 4 ℃. Primer screening process to Beirui Er, Sakhalin, the Daxinganling wild resource materials, screened 20 polymorphisms, clear and stable primer combinations amplified bands from 90 primer combinations. Amplified using primers screened 58 were 172 bands were amplified polymorphic bands of 143, 83.1% of the total strip, in which each pair of primers 5-12 strip. The genetic similarity coefficient between 0.53-0.96. 4, using the primers and screened after optimization system for PCR amplification of the varieties tested, electrophoretic bands of the form of 1/0 data conversion, select clear, in reproducing strong bands as a statistical object referred to as l, no The amplified bands Hutchison for the the UPGMA method clustering build tree and genetic diversity analysis. 58 resources are clustered into seven categories, the resources from the Russian regions clustered into one group, Yichun, Hisashi Boli's resources together, Altay, Xinjiang, the Daxinganling wild and Changbai Mountain wild resources separately clustering, each group Beirui Er, Bluebird, Blue spindle three cultivars and E3, E6, VIR together hybrid seedlings together as a class. Unknown No. 23 and Vladivostok resources together, No. 23 may be from the Vladivostok region resources or hybrids. Resources from the Northeast region is divided into four groups, be seen, the northeast region of the Blue Lonicera high genetic variation, as well as the northeastern region is one of the the blue the Lonicera genetic diversity center.
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