【作者】 王士娟；

【导师】 林英任；

【作者基本信息】 安徽农业大学 ， 微生物学， 2007， 博士

【摘要】 本文采用现代菌物分类技术,从形态解剖学、个体发育学、生态及生物学等方面对安徽黄山风景区的斑痣盘菌科(Rhytsmataceae Chevall.)的分类和物种多样性进行了较系统、全面的研究。同时在表型性状分类的基础上,基于ITS1-5.8S rDNA-ITS2区序列,构建了散斑壳属(Lophodermium Chevall.)系统发育树,探讨了散斑壳属种间及种内的遗传多样性及亲缘关系。以Darker(1967)、Cannon&Minter(1983,1986)、Johnston(1986)、Spooner(1991)、Kirk et al.(2001)等人提出的若干分类原则为主要依据,对自行采集和森林保护教研室保藏的650余号斑痣盘菌科标本进行鉴定与分类,共查明9属55种,其中除47个已知种外,还包括8个新种:冬青齿裂菌Coccomyces ilicis sp.nov.、苔草皮下盘菌Hypoderma caricis sp.nov.、树参散斑壳Lophodermium dendropancis sp.nov.、群生散斑壳L.gregarium sp.nov.、八角生散斑壳L.illiciicola sp.nov.、石楠生散斑壳L.photiniicola sp.nov.、斑茅舟皮盘菌Ploioderma sacchari sp.nov.及奇异小鞋孢盘菌Soleella mirabilis sp.nov.。编制了二叉式分种检索表。对新种提供了形态学特征的描述与图解以及寄主、生态和产果期等的记载,并加以讨论;对本人近期发表的新种提供了拉丁文特征简介。新种的模式标本保藏于安徽农业大学森保教研室(AAUFP)。本文在分类基础上对黄山区景区的斑痣盘菌从物种组成、寄主种类、寄生性及致病性等方面进行物种多样性研究,并分析了斑痣盘菌物种多样性形成的原因。其结果表明该风景区内斑痣盘菌科物种资源十分丰富,共发现55个种,即Lophodermium属21种、Coccomyces属18种、Hypoderma属5种、Rhytisma属4种、Soleella属3种,Bifusella、Hypohelion、Necoccomyces、Ploioderma属各1种;优势种为Coccomycesdelta、C.huangshanensis、C.multangularis、Hypoderma rubi、Lophodermium agathidis、Rhytisma punctatum等。该科菌物寄主范围十分广泛,共涉及21科(水龙骨科、松科、杉科、柏科、槭树科、五加科、冬青科、莎草科、杜鹃花科、壳斗科、樟科、金缕梅科、木兰科、防已科、木犀科、悬铃木科、蔷薇科、芸香科、山矾科、山茶科及禾本科)35属52种。它们是众多裸子植物、被子植物及少数蕨类植物上的强寄生物、兼性寄生物或腐生物,其中不少种类是维管植物的重要病原菌,有的甚至对黄山松、黄山杜鹃等特有树种的生长已构成严重威胁。采用改良的氯化苄法,获取了散斑壳属(Lophodermium Chevall.)29个菌株的基因组DNA。通过真菌通用引物ITS4和ITS5对rRNA基因的ITS1-5.8S rDNA-ITS2区进行了特异性扩增,扩增产物经纯化后测序,获得长600～1000bp的ITS区序列。以柔膜菌目的Helicodendron triglitziense(EF02923429)为外群,基于隶属20种的39个菌株ITS区序列(含29个自行测定和10个从GenBank下载序列),构建了散斑壳属的系统发育树。本系统发育树由3个进化分支构成。所有的针叶树生散斑壳形成进化分支Ⅰ,自举支持率为62%,它又由3个小的分支组成。5个Lophodermiumaustrale、6个L.conigenum以及L.pine-bungeanae、L.indianum和L.seditiosum菌株形成了自举支持率为54%的分支A。L.pinastri、L.macci、L.pini-excelsae和L.petrikii这4个种形成分支B,得到的自举支持率为74%。针叶树生散斑壳进化分支的基部是分支C,它由L.baculiferum、L.molitoris和L.kumaunicum组成,其自举支持率为71%。子囊果具唇的阔叶树生散斑壳菌株被聚在了一起而形成进化分支Ⅱ,得到了80%的自举支持率。所有子座覆盖层裂缝两侧均具严重碳化、无明显结构的黑色向内延伸物、覆盖层与基部层毗邻处为矩胞组织构成的内部基质的种被聚在一起组成了进化分支Ⅲ,它的自举支持率达99%。对该系统发育树的分析结果表明:1、散斑壳属种间关系与其寄主具很大的相关性,针叶树与阔叶树上的散斑壳属成员可能是亲缘关系较远的两大类群;2、位于进化树基部的进化分支Ⅲ与Ⅰ和Ⅱ组成的进化分支形成一个姊妹群,这说明散斑壳属很可能是多源的。3、寄主、生境和产地的不同会引起散斑壳属种内一定的遗传变异;4、子囊果的埋生位置、子座覆盖层与基部层的发育程度、唇细胞的有无等特征在散斑壳属种水平分类上具有重要的意义。更多还原

【Abstract】 Taxonomy and species diversity of Rhytismataceae from Huangshan scenic spot were systematically studied with the techniques of modern mycology.Moreover nucleotide sequences of the internal transcribed spacer(ITS)region of nuclear ribosomal DNA were used to infer phylogentic relationships within Lophodermium.More than 650 specimens were examined.According to the taxonomic principles of Darker(1967),Cannon & Minter(1983,1986),Johnston(1986),Spooner(1991),Kirk et al.(2001),9 genera 55 species of Rhytismataceae were identified.Among them, Coccomyces ilicis,Hypoderma caricis,Lophodermiurn dendropancis,L.gregarium,L. illiciicola,L.photiniicola,Ploioderma sacchari and Soleella mirabilis were new species. Keys were compiled for identification of all species.Morphological characteristics were described and illustrated,and hosts,ecology,fruiting period and discussions were provided for every taxon.The hosts of Rhytismataceae included 21 families 35 genera 52 species.Latin diagnoses of new species been published recently by the author were provided.Type specimens of all new species were deposited in the Forest Protection Laboratory of Anhui Agricultural University(AAUFP),Hefei,China.Species diversity of Rhytismataceae from Huangshan scenic spot were investigated in species composition,host range,parasitism and pathogenicity,and the reason of species diversity was analysed.The results showed that species resources of Rhytismataceae were abundant.55 species were identified including 21 species of Lophodermium,18 species of Coccomyces,5 species of Hypoderma,4 species of Rhytisrna,3 species of Soleella,and 1 species of Bifusella,Hypohelion,Necoccornyces and Ploioderma.The dominant species were Coccomyces delta,C.huangshanensis,C. multangularis,Hypoderma rubi,Lophodermium agathidis and Rhytisma punctatum and so on.Host ranges of Rhytismataceae were very broad that involved the following 21 families: Aceraceae,Aquifoliaceae,Araliaceae,Cupressaceae,Cyperaceae,Ericaceae,Fagaceae, Gramineae,Hamamelidaceae,Lauraceae,Magnoliacae,Menispermaceae,Oleaceae, Pinaceae,Platanaceae,Polypodiaceae,Rosaceae,Rutaceae,Symplocaceae,Theaceae and Taxodiaceae,35 genera,52 species.The members of Rhytismataceae were the parasites, facultative parasites or saprophytes of many Gymnospermae and Angiospermae plants,and less ferns,some of which were the pathogens of vascular plants,even harmed the endemic species of trees(Pinus taiwanensis,Rhododendron maculiferum ssp.anhwuiense etc.)in Huangshan scenic sopt. 29 genomic DNA from isolates of Lophodermium were extracted from mycelia following the improving method of benzyl chloride DNA extraction.ITS5 and ITS4 primers were used to amplify the ITS1-5.8S rDNA-ITS2 regions of isolates by the polymerase chain reaction(PCR).PCR products were sequenced after been purified. 600～1000bp ITS regions were obtained.Helicodendron triglitziense(EF02923429)of Helotiales was selected as outgroup. Thirty-nine sequences of 20 species,which were twenty-nine sequences been sequenced and ten sequences from Genbank,were analyzed.Phylogenetic tree of Lophodermium included 3 clades.All Lophodermium species on conifer trees composed CladeⅠwith 62%boostrap support,which involved 3 subclades:A, B and C.5 L.australe stains,6 L.conigenum stains,and L.pine-bungeanae,L.indianum and L.seditiosum formed Subclade A with 54%boostrap support.L.pinastri,L.macci,L. pini-excelsae and L.petrikii composed Subclade B,which had 74%boostrap support.The basal clade of conifer trees was Subclade C with 71%boostrap support,which included L. baculiferum,L.molitoris and L.kumaunicum.Lophodermium species with lips on broad-leaf trees were clustered together and formed CladeⅡwith 80%bootstrap support.CladeⅢwere composed of some species, whose ascomata have carbonaceous extending substance and internal matrix of textura prismatica in the edge of the ascomata.CladeⅢobtained very high bootstrap support (99%).The analysis of phylogenetic tree showed:1.Interspecific relation of Lophodermium were interrelated with their hosts.They were probably tow distant relation group between species on conifer trees and broad-leaf trees.2.CladeⅢin basal clade of the phylogenetic tree and another clade which consisting of Clade I and CladeⅡ,formed a sister clade, which suggested that Lophodermium was probably polyphyletic.3.The same Lophodermium species from different hosts and locations displayed certain genetic variability.4.Depth of embedding,degree of development of clypeus and lower wall and character of lips have important significance in taxonomy of Lophodermium.更多还原