鄂霍次克海天然气水合物区与东海内陆架泥质区沉积物古菌多样性研究

为研究鄂霍次克海天然气水合物区沉积物古菌、甲烷厌氧氧化古菌和硫酸盐还原细菌的多样性分布，我们以PCR技术为基础构建mcrA、dsrAB和古菌16S rRNA 基因文库。对所获得的序列进行系统进化和统计学分析发现：鄂霍次克海古菌类群主要为Marine Benthic Group D (MBG-D)、Marine Benthic Group B (MBG-B)、Marine Crenarchaeotic Group I(MG- I)，另外少量古菌16S rRNA基因序列为Anaerobic Methanotrophs 2c（ANME-2c），主要分布在LV39-25H岩心的表层沉积物中。LV39-40H岩心表层的古菌群落结构与其他六个层位古菌群落结构相比有着显著的差异。mcrA基因序列主要为催化甲烷厌氧氧化的古菌ANME-2（c和d簇），在所研究的各个层位的沉积物中均广泛分布。少量的ANME-1（a簇）发现于LV39-40H岩心表层以下的沉积物中。产甲烷古菌数目不多，集中分布在LV39-25H岩心200cm和LV39-40H岩心180cm的沉积物中。dsrAB基因文库分析表明硫酸盐还原细菌种类丰富，表层沉积物中硫酸盐还原细菌多样性最高。在两个岩心所有层位的沉积物中都有一定数量的克隆属于DSS簇，它们可能与ANME共生催化甲烷的厌氧氧化作用。总之，所有数据表明在鄂霍次克海天然气水合物区存在着较活跃的甲烷厌氧氧化作用，揭示了参与甲烷厌氧氧化作用的微生物群落结构和多样性。 为研究东海内陆架闽浙沿岸泥质区不同深度沉积物中古菌群落垂向分布特征，通过古菌16S rRNA 基因文库共得到473个有效克隆50个OTUs (Operational Taxonomic Units)。16S rRNA基因序列系统进化和统计分析发现古菌分别归属于泉古生菌（Crenarchaeota）和广古生菌（Euryarchaeota），其中以Miscellaneous Crenarchaeotic Group（MCG）为主，仅含少量的MBG-B、South African Gold Mine Euryarchaeotic Group(SAGMEG)、 ANME-3、MG- I和MBG-D。该泥质区沉积物可能存在由ANME-3催化的甲烷厌氧氧化作用，同源序列分析表明其古菌群落分布与周边环境有较大联系。UniFrac与沉积物环境因子分析表明该泥质区古菌群落垂向分布与沉积物有机质含量和粒度变化密切相关。 通过对比发现，鄂霍次克海天然气水合物区甲烷厌氧氧化古菌主要为ANME-2和少量的ANME-1，而东海内陆架泥质区甲烷厌氧氧化古菌仅为极少量的ANME-3；鄂霍次克海天然气水合物区广古生菌和泉古生菌数量各占一半，主要为MBG-D、MBG-B、MG-I。东海内陆架泥质区沉积物古菌序列主要为泉古生菌（MCG）。海域类型的不同以及有机碳含量等环境因子的差异可能是这两个海域古菌群落结构差异的主要原因.

Descriptions of Protospathidium serpens (Kahl, 1930) and P-fraterculum n. sp (Ciliophora, Haptoria), two species based on different resting cyst morphology

Protospathidium serpens (Kahl, 1930) is frequent in semiterrestrial and terrestrial habitats worldwide. Conventionally, all populations are considered as conspecific because they have very similar overall morphologies and morphometrics. We studied in detail not only the morphology of the vegetative cells but also the resting cysts using live observation, protargol impregnation, and scanning electron microscopy. These revealed a cryptic diversity and biogeographic pattern in details of the dorsal brush and cyst wall morphology. The cyst wall is spiny in the Austrian specimens, while smooth in the South African and Antarctic populations. Accordingly, P. serpens consists of at least two species: P. serpens (with spiny cyst wall) and P. fraterculum n. sp. (with smooth cyst wall); the latter is probably composed of two distinct taxa differing by the absence (South African)/presence (Antarctic) of a monokinetidal bristle tail in brush row 3, the number of dikinetids comprising brush row 1 (seven versus three), and the total number of brush dikinetids (29 versus 17). Protospathidium serpens is neotypitied with the new population from Austria. The significance of resting cyst morphology is discussed with respect to alpha-taxonomy and overall ciliate diversity.