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安徽常绿杜鹃花上斑痣盘菌物种多样性研究及散斑壳属两近似种的分子鉴定

Studies on Species Diversity of Rhytismataceae on the Evergreen Rhododendron Plants from Anhui and Molecular Identification of Lophodermium Conigenum and L. Australe

【作者】 顾婷婷

【导师】 林英任;

【作者基本信息】 安徽农业大学 , 森林保护学, 2010, 硕士

【摘要】 斑痣盘菌科(Rhytismataceae)及相关类群是世界菌物资源中的重要组成部分,广泛分布于世界各地。本文采用多性状综合分类的方法,以形态学特征、个体发育、生态习性、地理分布等表型性状为主要依据对安徽常绿杜鹃花上的斑痣盘菌进行较系统的分类与物种多样性研究。同时在表型性状分类的基础上,根据rDNA-ITS区序列,对散斑壳属两近似种进行分类鉴定,以明确二者种间亲缘关系及种内遗传差异。主要研究结果如下:1.以Darker(1967)、Cannon & Minter(1983, 1986)、Johnston(1986)、Spooner(1991)、Kirk et al.(2001)[10, 86, 14, 87, 74, 16]等人提出的若干分类原则为主要依据,对自行采集和安徽农业大学森林菌物研究室(AAUF)保藏的安徽省常绿杜鹃花上的干标本进行鉴定与分类。共查明隶属齿裂菌属(Coccomyces)、散斑壳属(Lophodermium)、新齿裂菌属(Neococcomyces)3属的7个种,即小齿裂菌(Coccomyces leptideus)、纠丝散斑壳(Lophodermium implicatum)、厚唇散斑壳(L. pachychilum)、红褐散斑壳(L. rufum)、坛状散斑壳(L. urniforme)、强壮散斑壳(L. validum)、杜鹃新齿裂菌(Neococcomyces rhododendri),其中坛状散斑壳为新种。编制了二叉式分种检索表,各种均附有形态解剖学特征的描述与图解以及寄主、生境的记载,并加以讨论。2.对针叶树散斑壳(Lophodermium conigenum)与其近似种南方散斑壳(L. australe)的形态学特征及生态习性等进行了比较研究,同时对这两个种8个菌株的rDNA-ITS区进行了PCR扩增和序列测定,结合GenBank中16个相关ITS序列构建了系统发育树。结果表明,L. conigenum与L. australe有着非常密切的关系。L. conigenum除子囊果形状、子座基部层、线纹和寄生性外,其余特征与L. australe基本相同。L. conigenum的ITS序列的G+C含量(51.0%)小于L. australe的G+C含量(54.0%)。在系统发育树中,此二种形成两个明显独立的分支,支持了依据形态学等表型性状的分类。二者种间及种内的遗传差异与寄主有较大的相关性,而与产地无明显关联。通过表型性状和ITS序列的分析与比较,可准确地将L. conigenum与L. australe鉴别开来。

【Abstract】 Rhytismataceae and its related groups are an important part in fungi resources of the world, which are distributed worldwide. Some members of Rhytismatales on the evergreen Rhododendron plants from Anhui were systematically studied by the synthetical classification means on multiple character and according to phenotypic character, such as morphology, ontogeny, ecology, and distribution. At the same time, Lophodermium conigenum and related species L. australe were identified and classified to definite the relationships in interspecies and genetic diversity in intraspecies on the basis of Phenotypic character and the sequences of rDNA-ITS region. The main result is as follows:1. Dry specimen on the evergreen plants of Rhododendron in Anhui from own collection and the Herbarium of Forest Fungi of Anhui Agricultural University (AAUF) were identified and classified according to the taxonoic principles of Darker (1967), Cannon & Minter (1983, 1986), Johnston (1986), Spooner (1991)and Kirk et al. (2001). 7 species of Rhytismatales were studied, including C. leptideus of Coccomyces, L. implicatum, L. pachychilum, L. rufum, L. urniforme and L. validum of Lophodermium, N. rhododendri of Neococcomyces. L. urniforme is a new specie 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. Morphological characters and ecological habit, etc. of Lophodermium conigenum and related species L. australe were compared; besides, rDNA-ITS region of eight strains of the two species was amplified and sequenced, and phylogenetic trees were constructed based on these sequences as well as sixteen related ones in GenBank. Phenotypic characteristics of L. conigenum are remarkably similar to those of L. australe, except shape of ascomata, stroma basic layer, zone lines and parasitism. G+C content of L. conigenum (51.0%) is less than that of L. australe (54.0%). The phylogenetic analysis shows the two related species are sharply classified into two independent groups, which are consistent with the classification based on phenotypic characters. The interspecific and intraspecific genetic diversity is closely related to hosts, but not to distribution. The comparison analysis of phenotypic characteristics and ITS sequences could be used to distinguish L. conigenum and L. australe accurately.

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