Studies On Taxonomy Of Cauline Species Of Rhytismatales And Molecular Biology Of Lophodermium On Conifers

Rhytismatals is an important part in fungi resources of the world. Systematics of Cauline species of Rhytismatals was studied from morphology, development biology, ecology, distribution,etc. Twenty-two Lophodermium strains on conifiers were charaterized by ISSR-PCR reaction with 10 random primers whose gel electroersis diagrams can be amplified repeatedly and clearly. By the comparison of ISSR analytic results and the phenotypic characters, indicated relationships of some intraspecific and interspecific strains of Lophodermuim on conifiers were affirmed. The main result is as follows:(1) specimens from the south and west mountains of Anhui province and Anhui Agricultural University Forest Protection (AUUFP) were identified according to the taxonoic principles of Darker (1967), Cannon & Minter (1986) and Kirk et al. (2001) .8 species of Rhytismatales were studied, including A. Rugosa of Ascodichaena, L. puerariae sp. nov., L. caulinum sp. nov., L. camelliicola and L cephalotaxi of Lophodermium, C. leptideus and C. Taiwanensis of Coccomyces, H. Stephanandrae of Hypoderma. A. Rugosa is a new record species in china, L.puerariae sp. nov. and L.caulinum sp. nov. are new species among them. Dichotomous key to these species were complied. Morphological characteristics were described and illusteated, and habits, hosts and distributions wre recorded for the species.(2) Template DNA of 22 Lophodermium strains and on conifers were extracted by Benzyl Chloride. The ISSR-PCR amplified system was optimized from aspects of the concentra- tions of DNA templates, primers, Mg²⁺, dNTPs and Taq DNA polymerase, and annealing temperature. The results showed that the optimal ISSR-PCR system were suitable for Lophodermium. as follows: template DNA (45ng·μl^-1) 2μl, primer (10umol·L^-1) 0.6μl, Mg²⁺(25mmol·L^-1)1.5μl, dNTPs (10mmol·L^-1)0.3, Taq DNA polymerase (5U·μl^-1) 0.3μl, 10×Buffer 1.5μl, ddH₂O 8.8μl in 15μl reaction system. 22 Lophodermium strains on conifiers were amplified using inter-simple sequence repert (ISSR)markers, which amplified 167 sites including 160 polymorphic sites, accounting for 95.8%. Its genetic distance cluster figure was constracted using UPGMA method. From the clustering figure we can find that the distance of genetic ranged from 5.93 to 7.41. 22 strains of Lophodermium on conifiers were classified to 5 groups. By the comparison of ISSR analytic results and the phenotypic character, The strains of intraspecies and similar interspecies can cluster very well, and the strains of different species also can distinct quite well. From the results we can conclude that ISSR technique has good applicability in the study of relationships of intraspecies and similar interspecies .At the same time ,we can confirm the importance of some morphological character such as the embedded position of ascomata, stromata, asci, ascospores, paraphyses and the category of hosts on the classification of Lophodermium.