南中国海纤毛虫原生动物的区系与多样性研究

Researches on geographic distribution and diversity of ciliated protozoan in marine unknown environments, especially representative habitats, is one of most weakness and blank area in the current international ciliatology; furthermore, studies on tropical and subtropical marine taxa are particularly lacking. Previous work performed by the applicant group in last decades was mainly focusing on the groups in northern China seas while those in southern China waters remain largely unknown. The application project will target at long-term insufficiently explored, free-living forms in various Guangdong coastal mangrove wetlands and other biotopes in south China Sea, and focus on geographic fauna and distribution, community structure, taxonomy and systematics, and ecological function and change. Main research contents include: 1). To carry out studies on species identification and descriptions, spatial and temporal distribution, ecological characteristics of ciliates in mangrove wetlands along the coasts of southern China, and developmental mode of representative forms. It aims to fill in the gaps of study in diversity of ciliates in tropical – subtropical oceans at gene, species and ecosystem levels by means of classic, modern taxonomy and ecology; 2). To construct DNA library of many molecular information missing morphospecies; 3). To sequence about 2000 molecular mark genes (e.g. SSU rDNA, 5.8S, LSU rDNA, alpha-tubulin etc) of unknown taxa and submit data to the international GeneBank; 4). To carry out researches on gene and metagenomic barcoding of representative species based on the above marker gene screening and to perform ontogeny of some evolutionarily important taxa; 5). To complete phylogenetic analysis and system reconstruction of six classes (e.g. Haptoria, Karyorelictea, Phyllopharyngean) and ca. 15 orders of ciliates on the basis of analyseis of molecular and ontogenetic information.

海洋未知环境中纤毛虫的分布和区系研究为当今原生动物学领域中突出的薄弱环节之一，而全球范围内热带-亚热带水体中的研究尤为欠缺，此现状构成了海洋生物多样性、环境变化、生物地理学以及微食物网内能量循环与评估等研究的瓶颈。本项目以我国广东、海南沿海的自由生纤毛虫为目标，聚焦在地理区系、多样性、分子及生态学信息构建等。主要工作包括：1）开展对红树林等典型生境中纤毛虫形态种鉴定、新阶元描述、时空分布、生态学-区系资料库建立及关键类群的细胞分化模式研究，填补热带－亚热带海域中纤毛虫物种-生态系多样性研究的空白；2）构建和完善国际间分子信息缺失的海洋类群的DNA种库；3）完成迄今空缺类群的标记性基因的测序和提交；4）在分子信息采集的基础上，开展代表性类群基因条形码与宏基因条形码的研究；5）在分子与个体发生学信息分析的基础上，完成对钩刺类、叶咽类等6个纲级、约15个目级类群的系统发育分析与系统数的构建。

本项目以我国广东、海南沿海各类生境中自由生纤毛虫的多样性为目标，围绕物种鉴定、个体与系统发育、基因条码和遗传信息库构建、生态分布、生态模型构建等核心内容开展了系列研究，全面、超额完成了预定任务。主要成果包括：1）在大量采集、分类、鉴定基础上，完成了对长期存在混乱、不详属种的重厘定、重定义、纤毛图式资料的建立并在国际刊物上发表、建立了1新科、4新属、35个新种；2）完成了对30种腹毛类代表种的细胞发生模式的刻画；3）完成了170余形态种的DNA建库、150余种SSU rRNA等标记基因的测序及向GenBank数据库的提交；完成了对7个代表种的常规组学及96个类群代表种的单细胞组学数据的测定和建库；4）基于多基因信息及系统基因组学信息完成了对腹毛类、游仆类、盾纤类、管口类等阶元的系统探讨与修订，并在此基础上系统地重建和梳理了全面覆盖纤毛门内55个目级阶元的系统进化关系；5）完成了8个区段的条形码基因筛选、58种1100余条条形码数据库的构建及宏基因条形码在寡毛类纤毛虫中的应用；6）从种群（动力学特征）、细胞（生理生化酶活性）和基因等生物学水平上，研究了环境污染物等环境因子对原生动物的生态毒性效应及其响应模式。.上述成果先后形成国际主流刊物文章53篇，包括在《Mol Ecol Resour》、《Proceedings B》、《Mol Phylogenet Evol》等领域重要刊物上发表论文12篇，出版专著一部《Atlas: Ciliates found from south China sea》（Springer出版社）；分子系统学分支的成果获教育部（2019）自然科学成果二等奖。同期成果还包括培养毕业博士生13人、硕士生21人，毕业生中获“山东省优秀博士学位论文奖”3人、“山东省优秀硕士学位论文奖”1人。