Knowledge Resource Center for Ecological Environment in Arid Area
| 中国叶上衣属分类及糙聚盘衣物种资源生物学研究 | |
| 其他题名 | The research on taxonomy of Strigula and species resources of Glypholecia from China |
| 蒋淑华 | |
| 出版年 | 2017 |
| 学位类型 | 博士 |
| 导师 | 魏江春 |
| 学位授予单位 | 中国科学院大学 |
| 中文摘要 | 由于地衣多样性和系统性研究是自然界地衣多样性与地衣资源研发之间的桥梁，本研究结合《中国地衣志》编前研究任务进行叶上衣属地衣物种多样性与系统性生物学并结合沙漠生物地毯方向进行糙聚盘衣物种资源生物学研究。通过对采集自我国热带、亚热带地区的800余份叶上衣属地衣标本，在形态学、解剖学、化学、地理分析和系统发育学综合分析的基础上研究，得出以下结果：获得了该属ITS rDNA序列430条，SSU rDNA 65条，LSU rDNA 87条，TEF1-α 120条，RPB2 71条。通过多基因联合的系统发育学分析，同时与表型相结合，将原来的叶上衣属重新划分为6属，除狭义的Strigula属外，其中3属为新属，另外2属为恢复原叶上衣科的Raciborskiella属和Phylloporis属。基于LSU和SSU序列，用更多的分子数据进一步证明了叶上衣属应置于座囊菌纲独立的目叶上衣目。本研究从叶上衣科3个属的14个物种中扩增得到28条LSU全长序列（3071–4352 bp）。以该属模式种Strigula smaragdula为模板，在LSU rDNA全序列中找到了其中10个结构区。通过核苷酸序列的比对分析，鉴定出狭义叶上衣属6条特有的核苷酸序列。本研究记载和详细描述了中国叶上衣属地衣30种，其中包括14个新种和6个中国新记录种。对复合种Strigula smaragdula和S. nitidula进行了研究，并支持S. difformis和S. hypothelia分别提升为种的水平，不再作为变种和变型处理。采用LSU和TEF联合基因片段进行系统发育学分析发现石生种类 Strigula jamesii形成的分支与叶上衣科另外2属Flavobathelium和Phyllobathelium嵌合在一起，表明该种并不是真正意义的Strigula成员，本论文将其处理为恢复原属Geisleria的成员。对糙聚盘衣属物种资源进行了相关研究，研究结果如下：通过对地衣型真菌的抑菌活性初筛发现，糙聚盘衣属两种的共生菌甲醇浸提液具有抑制革兰氏阳性菌和阴性菌的活性。通过与地衣体次级代谢产物丰度比较发现，糙聚盘衣属两种共生菌次级代谢产物丰度均高于其对应地衣体。从Glypholecia scabra分离培养的共生菌中通过各种柱层析技术分离获得了7种代谢产物，利用各种现代波谱技术已解析6个化合物的结构，包括5种聚酮类化合物和1种含硫的二酮哌嗪类化合物，均为首次在地衣中报道。同时，从该属另一种Glypholecia sp.纯培养的共生菌中分离获得1个萘醌类化合物和2个生物碱类新化合物。目前，正在进行新化合物单晶培养，通过X-Ray单晶衍射的方法确定其绝对构型、并对其相关活性如细胞毒活性、抗病原真菌等进行测试。 基于目前的研究结果表明，分离培养的地衣共生菌能产生多于共生状态地衣体的、并具有一定生物活性的次级代谢产物，具有开发优势和价值，可作为挖掘真菌次级代谢产物生物合成潜力的有力材料。 |
| 英文摘要 | The research on biology of diversity and systematics of lichens is a bridge between species diversity and species resources. On the basis of the diversity and systematics of Strigula for the project of the book “Lichens in China”, species resources of Glypholecia was also included in this thesis for the Desert Biological Carpet Engineering.Strigula Fr. are belonging to Strigulaceae, Dothideomycetes, Ascomycota, Fungi. Mostly are found on leaves in tropical to subtropical regions. Strigula has been previously circumscribed based on morphology, and species delimitation in this group generally lack comprehensive analysis. This is the first time to focus on the systematic studies of the group by the combination of phenotype and genotype. The main results and conclusions presented in this thesis are as follows:More than 800 foliicolous Strigula specimens were examed in this study. A total of 430 ITS rDNA sequences, 65 SSU rDNA sequences, 87 LSU rDNA sequences, 120 TEF1-α sequences and 71 RPB2 sequences were obtained. We add, for the first time, molecular data to elucidate phylogenetic relationships at the species level in this genus. Multi-locus phylogeny analysis including ITS, SSU, LSU, TEF1-α, RPB2, showed Strigula species clustered in six clades. It indicates that Strigula can be divided into six genera based on morphological and molecular evidence, including three new genera, with the revival of Raciborskiella and Phylloporis in Strigulaceae.Molecular phylogenetic approach using LSU rDNA and SSU rDNA present an updated phylogeny of Strigulaceae with more molecular data, demonstrating that the lineage is best recognized in a separate order Strigulales in Dothideomycetes.In this study, twenty-eight LSU full-length sequences (3071–4352 bp) were amplified from fourteen species of the group. As an example of Strigula smaragdula, the full length LSU rDNA can be divided into 10 structure region in terms of the different evolutionary rates in the genus Strigula s.str., and the diagram of Strigula smaragdula LSU rDNA gene structure was built. There were six characteristic sequences of Strigula s.str. in the region of the LSU rDNA sequences.Thirty species of this group were found from China, including fourteen species new to science and six species new to China. The broad concept of Strigula smaragdula and Strigula nitidula indeed represent species complex, rather than one species. According to our study, some minor morphological traits are diagnostic characters for segregating different species. Taking molecular difference into consideration with morphology, here we suggest the revival of the dinstinct species Strigula hypothelia and Strigula difformis, rather than a form or a variety of S.nemathora.The saxicolous species Strigula jamesii nested with Flavobathelium and Phyllobathelium in Strigulaceae in phylogenetic analysis. This suggested that Strigula jamesii should be excluded from Strigula, and revived into another independent genus Geisleria. However, other specimens and sequences of Geisleria are needed for more research.In addition, the secondary metabolites of lichenized fungi Glypholecia was also studied. The main results are as follows: In order to explore the antibacterial activity of lichenized fungi, the mycobiont isolated from desert lichens were chosen for antibacterial activity screening. Two species, Glypholecia scabra and Glypholecia sp., showed inhibition of three bacteria. More compounds are detected in the mycobiont by TLC than in corresponding lichens.Seven compounds were isolated from the mycobiont of Glypholecia scabra, and. six of them have been elucidated the structure, including five polyketides and one Sulfur-containing diketopiperazine. Bacilosporin D and two new alkaloid compounds were isolated from another species Glypholecia sp., but the absolute configurations of the two new compounds need to be determined by X-ray crystallography in further study. It is also necessary to test the activity of the two new compounds. The research indicates that the mycobiont can produce more secondary metabolites with biological activity than lichens. Screening of compounds with new structure and biological activity will improve the utilization of desert lichen resources, which are worthy of in-depth and systematic study. |
| 中文关键词 | 分类学 ; 表型 ; 基因型 ; 共生菌 ; 次生代谢 |
| 英文关键词 | Taxonomy Phenotype Genotype Mycobiont Secondary metabolites |
| 语种 | 中文 |
| 国家 | 中国 |
| 来源学科分类 | 微生物学 |
| 来源机构 | 中国科学院微生物研究所 |
| 资源类型 | 学位论文 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/287928 |
| 推荐引用方式 GB/T 7714 | 蒋淑华. 中国叶上衣属分类及糙聚盘衣物种资源生物学研究[D]. 中国科学院大学,2017. |
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