Sea Buckthorn Cultivars Identification Based On Rapd Markers

Sea buckthorn (Hippophae rhamnoides L.) is a kind of plant of sea buckthorn in elaeagnacae, it hasecological benefits, economic benefits, social benefits and many valuable features (Welsh et al.,1990).Sea buckthorn is Pioneer species of the Three-North protect forest in china which has windbreak andsand, soil and water conservation, slope revetment, soil building, enhancing soil fertility and otherecological functions, sea buckthorn is also a kind of medicinal fruit trees, value and high nutritionalvalue medicinal, it spreads a large area in China as the cultivation of fruit trees. Because sea buckthornhas varieties type, a wide distribution and high degree of hetero zygosity of the genome, it forms newspecies and clones with ecological adaptability quite different and huge number in long term the naturalevolution and the process of inter specific hybridization, in the past the identification method is difficultto research and analysis accurately. Emerging molecular marker technology to overcome manyshortcomings of the morphological methods, especially the RAPD technique with simple, fast,non-isotopic pollution, and less DNA polymorphism, and it is widely used in cropgermplasm andgenetic diversity analysis. That is a very important significance for Understanding of the geneticrelationship between all varieties of sea buckthorn on the study and utilization of plant germplasm.In this experiment, the author chooses11-nucleotide random primers RAPD amplification of Seabuckthorn constructed according to specific bands corresponding map relationships, rapid identificationby Manual the Cultivar Identification Diagram (MCID), in order to distinguish varieties. The method isconvenient, fast, accurate, and seven PCR can be to separate67sea buckthorn cultivars at the molecularlevel, cultivar identification diagram is more intuitive than the clustering tree, according to cultivaridentification diagram can be find to distinguish any two species of primers. The method can achieveearly identification of seedlings, with a wide range of versatility in other species, it has a positivemeaning for the identification of sea buckthorn germplasm resources and promotion of sea buckthornindustry continued development of.According to the experiment result, extracted by two methods of DNA CTAB method and SDSmethod, the experimental results show that, the two methods can extract the genomic DNA more, butcompared with CTAB method, better results. Through two times of continuous gradient PCR striptemperature as primer annealing temperature, the experimental results show that, this experimentchooses41℃as the annealing temperature is appropriate; in order to make RAPD technology,designed a pair of primers for PCR reaction of11bases in length, selected with random amplifiedpolymorphism of high, good stability, higher characteristic; through the band contrast screening,screening out the study consisted of11nucleotides with polymorphism and stability characteristics ofthe random primer seven, namely67seabuckthorn varieties in7separate PCR can at the molecular levelarea.Method of MCID amplification products of the RAPD not only makes the purity of seabuckthornseedlings were identified, but also improve the accuracy of screening of germplasm resources, broaden the research method of identification of varieties of fruit bodies of knowledge, the method can beapplied in variety property protection seedling management support, germplasm resources management,nursery stock enterprises, is research and further study of this topic can also target identification ofvarieties of male and female.