POD Isoenzyme And RAPD Analysis Of Longan(Dimocarpus Longana L.) Genetic Resource

Longan(Dimocarpus longana Lour.) is one of the most important subtropical fruit trees in Fujian province. But the development of Longan resources are restricted severity for knowing little about the genetic background. The genetic diversity and the genetic relationship among cultivars of longan were studied by using the random amplified polymorphic DNA technique (RAPD)and the analysis of POD sioenzyme in this paper. The major results obtained from the study are summarized as follows:1. The method of POD isoenzyme extract the system of electrophoresis were optimized. According to the factors which effect the activity of POD isoenzyme and number of the bands in the gel such as the fresh and tender of the material, the component of the extracting buffer, the methods of electrophoresis and dyeing and so on, the method fit for extracting the POD isoenzyme and its system of electrophoresis were established after iterative experiments. The longan leaves age from 3 to 6 months were the best material for POD isoenzyme. The 0.05mol/L PBS (pH7.5) adding 0.1%Triton was the adaptive extracting buffer. PAGE was chosen for electrophoresis, methods of acetic acid and benzidine for dyeing.2. An analysis of the genetic relationship of longan resources using the analysis POD isoenzyme. The whole zymogramic includes 17 bands and was divided into 4 regions. Regionâ…  and â…¢ are active regions, the bands of the enzymes in the two regions were more fixed and active. The activity of the enzymes in Region â…  and â…¢ were weaker. The second and third bands (whose RF were respectively 0.220 and 0.253) were the fundamental bands of longan. Using clustering analysis by POD isoenzyme, the 95 genetic resources of longan were divided into 2 types and 7 subtypes.3. Establish the method of longan genomic DNA extraction from leaves. Some secondary metabolites such as polyphenols, polysaccharides and pigments were rich in the leaves which are easy to form complex with DNA. The complex was insoluble and could inhibit theactivity of Taq...