Analysis And Study Of RAPD-PCR On Lactarius Deliciosus And Isolated Strains

It' s the first time for this dissertation to analyze Lactarius deliciosus and isolated strains with RAPD-PCR. Four different DNA extractions which are improved after selection are used in this research. The DNA of Lactarius deliciosus and isolated strains are extracted, Lactarius deliciosus come from LiLing mountainous area of china, and the isolated strains which are cultivated in shaking flask are obtained from tissue culture of Lactarius deliciosus. A highefficiency DNA extraction, synthetical extraction, which can be used for Lactarius deliciosus is obtained. A series of searches are experimented about the conditions of RAPD-PCR , with which the heredity differentia between Lactarius deliciosus and isolated strains are analysed on the molecule level. Moreover, the heredity stability of isolated stains is analysed. Twenty five primers whose DNA fingerprinting patterns are clear and repeated highly are selected from one hundred 10bp random primers. 185 loci are obtained in the amplification for the experimental samples with these 25 primers, 7.4 loci per primer averagely. The similarity coefficients are calculated on the base of the six samples' DNA fingerprinting patterns. The similarity coefficients which range from 0.8000 to 0.9784 are high between Lactarius deliciosus and isolated strains. The similarity coefficients between strain 2, strain 3 and sample 1 are all over 0.9351. Especially, the similarity coefficient of sample 1 and sample 2 is higher to 0.9784. Strain 4 is the second generation from Lactarius deliciosus, and the similarity coefficient of sample 1 and sample 4 is 0.8541. The similarity coefficients between Lactarius deliciosus to strain 5 and strain 6, which are isolated and cultivated from generation to generation for third times, are 0.8000 and 0.8108 respectively. These similarity coefficients are high. The conclusion that Lactarius deliciosus and isolated strains are the same species can be drawn. Furthermore, cluster analysis(UPMGA) is applied in the light of the six samples' similarity coefficients. The result of the cluster analysis shows that six samples cluster together at similarity coefficient 0.8, more or less. The initial conclusion that Lactarius deliciosus and five isolated strains are the same species can also be obtained. Both in the similarity analysis and the cluster analysis, the conclusion that Lactarius deliciosus and isolatedstrains are the same species can be indicated. What' s more, the heredity differentia and heredity stability of the isolated strains are analysed according to the similarity confident and cluster analysis, the analysis indicate that heredity stability of the isolated strains are relative. A high efficiency DNA extradition for mycorrhizal fungi is obtained. In addition, a new, fast, convenient and scientific approach which is used in the discriminate for the isolated strains of Lactarius deliciosus comes into being. The very convincing references are provided in this dissertation to study the molecule biology of Lactarius deliciosus further. This paper lays a foundation for the research and exploiture on Lactarius deliciosus.