Studies On The Microtubular Organelles And Extrusive Organelles Of Onychodromopsis Sp(Hypotrichida, Ciliophora)

Hypotrichous ciliate are highest group in ciliates, it's most typical groups with characteristics namely complicated ciliature apparatus. These ciliature apparatus consist of microtubular organelles and the base associated microtubules of these ciliatures:adoral zone of membranelles (AZM) and its base associated microtubules (membranelle brackets, membranelle associated microtubules), undulating membranes (UM) and its base associated microtubules, frontal-ventral-transverse cirri (FVTC) and its base associated microtubules(anterior longitudinal microtubules (ALM), posterior longitudinal microtubules (PLM), transverse microtubules (TM)), marginal cirri (MC) and its base associated microtubules(ALM, PLM, TM), caudal cirri (CC), dorsal kineties (DK). The main ingredients of ciliature are microtubulins, therefore ciliated protozoa are common materials for research of microtubular organelles and microtubulins of ciliates.According to different characteristics on morphology and morphogenetic, ciliated protozoa can be classified into different groups. In taxonomy, Pleurotrichidae is a transition gronp between Urostylidae and Oxytrichidae. For Pleurotrichidae have low category and quantity, and few related researches, its classification are out of order. In this study, by using direct-fluorescent labeling,TEM,SEM,SDS-PAGE,biochemical extraction, the strucutures and functional components of microtubular organelles, morphological structure of extrosomes, and microtubulin were discussed. Expect to provide materials and evidence for taxonomic research of Pleurotrichidae. Results are as follows:1 The morphology and morphogenetic of the ciliature microtubular organelles in Onychodromopsis Sp The morphology and morphogenetic of the ciliature microtubular organelles in the ventral cortex of Onychodromopsis Sp were visualized using FLUTAX direct fluorescent labeling. In the front of cell, the AZM possesses 30～50 adoral membranelles; UM possesses 2 rows of cilia; FVTC distribute into 8 frontal,5 ventral and 5 transverse cirri; 2 left marginal cirri (LMC) and 3 right marginal cirri (RMC),6 DK rows; Developed ALM and PLM of FVC, and developed ALM of TC, the first four ALM of TC extend forward together into a triangle structure. TM of LMC are extreme underdevelopment. During morphogenesis, the new MC primordium first originate from the outest RMC, and then MC primordium presents in the front and the middle part of th degenerated site if the old marginal cirri. On the features of its ciliature and the base-associated microtubules, this species qualifies to be the genus Onychodromopsis. However, the morphogenetic study shows the originating position of oral primordium of the opisthe and differentiation of the marginal rows were different from other related species Onychodromopsis flexilis, this species was probably another species in the genus Onychodromopsis.2 The ultrastructure of cytoskeleton in Onychodromopsis SpThe results of TEM showed that ultrastructure of microtubular cytoskeleton of Onychodromopsis Sp. It showed that a layer of alveolar membrane beneath the pellicle; ciliary shafts possess "9+2" doublet microtubules; and basal bodies of the cirri are also linked by different kinds of connections. Each membranelle of AZM has two types of compositional model:2 rows of cilia and 3 rows of cilia; A layer fiber is cover around FCB, and lots of microtubules send forth from FCB, developed ALM. There has 3-5 microtubules layers beneath the pellicle. The pellicle closed to AZM has thicken microtubules layers, which was consisted of hundreds of layers. During morphogenesis, Macronucleus appeared replication band, it's a homogeneous structure. Reasults shows compared with others ciliates, Onychodromopsis Sp have different microtubules layers beneath the pellicle and republication band of Macronucleus. These characters are correpond with evolutionary discipline from Urostylidae, Pleurotrichidae, Oxytrichidae to Euplotidae. To provide the mateials for taxonomic research of Pleurotrichidae at sub-microscopy level.3 The ultrastructure of the extrusomes in Onychodromopsis SpBy using SEM and TEM, the ultrastructure of extrusomes in Onychodromopsis Sp to be observed. SEM shows there has granulate structure on surface of the body, and less in vicinity of AZM; TEM shows that extrusomes distributed in the cortical cytoplasm of Onychodromopsis Sp Mature organelles are located beneath pellicle,the top part of extrusome have low density fibrous matter, even appear cavity; In the process of morphogenesis of extrusomes, it appear concentric circles structure with high-low electron-dense matter. And vesicle extended to be divided in two parts, which also has concentric circles structure. The morphology and morphogenetic of extrusomes indicated this extrusome was probably a kind of mucocyst, the divide of its probably a new way of origin. And this extrusome might be a protective or defensive response to the environment changes, irrelevant with predatory behavior.4 The protein composition of Onychodromopsis Sp and cloning analysis of gramma-tubuliln gene fragment from Onychodromopsis SpBy using the methods of SDS-PAGE analyse the protein composition of Onychodromopsis Sp, meanwhile cloning of gamma-tubulin gene fragment from Onychodromopsis Sp. The result of SDS-PAGE shows that the molecular weight of protein range from 66.2-116kD and 35-45KD was rather higher, the protein composition of 30kD,20kD, and 16kD bands come to 7.0-7.9%; The result of clonging showed that the molecular weight of gamma-tubulin is 1000bp approximately. It demonstrated that Onychodromopsis Sp has different characters in the protein composition and gamma-tubulin gene fragment.