| Rhytismataceae is an important part in fungi resources of the world.Systematics of part of species of Rhytismataceae was studied from morphology,development biology, ecology,distribution,culture characters etc.with the techniques of modern mycology.And phylogenetic relationships of intraspecies and similar interspecies of Lophodermium and its two similar genera(Ploioderma,Soleella)strains on conifiers were studied.It is clear further to the phylogenetic relationships among these groups.Some samples from the mountains of south and west Anhui and were systematically studied with the techniques of modern classification.Morphology structure and development biology of these fungi were roundly examined according to taxonomic principles of Darker(1967),Korf(1973),Cannon & Minter(1986),Kirk et al.(2001). 8 species among 4 genera were identified.Among the taxa there were 3 new species including Hypoderma querci sp.nov.,Lophodermium castanopsis sp.nov.and Coccomyces brevisporus sp.nov.;the studies of 5 known species involved in adding characters of morphology,development biology,new record of host and geographical distribution.Morphological characteristics were described and illustrated,and hosts and distribution were recorded for all the species,and their similar groups,parasititism and nosogenesis were discussed.Dichotomous key of the species was compiled.Living materials of the fungi from the mountains of south and west Anhui were isolated by tissue isolation technics,and 10 strains of Lophodermium and its similar genera on conifiers were obtained.Culture characters were studied by observing,recording and screening the chromophotographs of pure cultures periodically.The result showed that there were definite difference in growth rate,shape,structure and color of colonies and mature period of fruiting bodies in MA culture media.Few of the species formed pycnidia, and none produced ascomata during the culture.Only very few of the strains produced coloring matter that make culture media colored.Template DNA of 26Lophodermium strains and 2 Ploioderma destruens and Soleella chinensis strains on conifiers were extracted by Benzyl Chloride.The RAPD-PCR amplified system was optimized from aspects of the dosage of template DNA,prime,Mg2+, dNTPs and Taq enzyme,and anneal temperature and time,elongation temperature and time, cycle number.The amplified diagrams were quite well.The final reaction system(25ul) included ddH2O 14.2ul,10×Buffer 2.5,Mg2+(25mmol.L-1)2ul,dNTPs(10mmol.l-1)1.5ul, primer(12.5umol.L-1)0.6ul,Taq enzyme(5U.ul-1)0.2ul,template DNA(100ng.ul-1)1ul. Its amplified procedure was pre-denature at 95℃for 5min.cycle 50 times,and every cycle included denature at 94℃for lmin,anneal at 36℃for 100s,elongation at 72℃for 140s. And the latest cycle elongation time is 10min.24 Lophodermium and 2 related genera strains on conifiers were amplified by the optimal procedure,which amplified 111 sites including 104 polymorphic sites,accounting for 94%.The length of most amplified fragments ranged from 250 to 2000bp.Its genetic distance cluster figure was constracted using UPGMA method.From the clustering figure we can find that the distance of genetic ranged from 1.98 to 5.35.26 strains of Lophodermium and 2 related strains on conifiers were classified to 6 groups(A,B,C,D,E and F)in the genetic distance 4.91.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 RAPD technique has good applicability in the study of relationships of intraspecies and similar interspecies.Compared with phenotypic character and RAPD analysis results,we can conform the importance of the phenotypic characteritics such as zone lines,ascomata and hosts etc.in the classification of intraspecies and similar interspecies of Lophodermium on conifiers. And the type of ascopores was proved being important in distinction of Lophodermium and its similar genera on conifiers from another ascept.
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