伪尖毛虫Oxytricha fallax的形态学研究

伪尖毛虫Oxytricha fallax系原生动物门、纤毛虫纲、腹毛目、尖毛虫科的一种常见的单细胞动物。一般认为分裂间期的伪尖毛虫具有两个长圆形大核和两个圆球形小核。虫体腹面生有八根额棘毛、五根腹棘毛及五根肛棘毛,周缘左右两侧还生有缘棘毛,背面则有五列背触毛。 伪尖毛虫是原生动物中高度进化的类型,大、小核有显著的分化,细胞表面的纤毛小器官复杂而又明显,因此它是用以进行研究的一种良好材料。对它的核器与纤毛小器官以及它们在无性分裂过程中的发生和变化进行认真的探讨,可以使我们较详细地掌握该虫的形态特征,同时也能为进一步的研究提供一些资料。     

背联体贝尾棘虫大核摘除、移位后的调节现象

人工背联体贝尾棘虫是两个正常贝尾棘虫以背部相联接的联体,它具有两个能进行摄食的口,有两套大核和小核。由于背联体两测的细胞质大小相似,无性生殖是同步发生的,所以具有正常的生殖能力(Tchang and Pang,1979)。我们在这种“背联体”的培养中观察到它的大核的多种调节现象(Tchang and Pang,1979),认为这是细胞核质关系协调和平衡的例证。我们考虑,如果在人为干扰的情况下,这种“背联体”又会发生什么变化呢?本文报告对人工背联体贝尾棘虫进行摘除大核和将大核移位所引起的变化过程,从中探讨细胞核质关系及其作用问题。     

氘水诱发伪尖毛虫核与毛基系的变异

本文以伪尖毛虫(Oxytricha fallax)为材料,进行不同浓度的氘水处理。实验表明,氘水诱发伪尖毛虫大核变异,随氘水浓度增加而畸变率上升,但畸变幅度不大,移入正常环境中即恢复正常。氘水诱发伪尖毛虫毛基系发生变异,放入正常环境后观察73天(约150代)仍未复原,证明氘水使伪尖毛虫的毛基系发生能遗传的“傍徨变异”。     

尖毛虫属Actin Ⅰ、α-TBP和DNA pol α乱序基因的模式研究

研究旨在对尖毛虫属内现有物种的3种乱序小核基因结构进行比较,探讨其乱序模式。于湛江湖光红树林水域中采集到一个尖毛虫属物种Oxytricha sp.（ZJ）,成功扩增了其肌动蛋白Ⅰ（ActinⅠ）、端粒结合蛋白（α-TBP）、DNA聚合酶α（DNA pol α）3个乱序基因的完整大核基因序列和完整/部分小核基因序列,并结合已有资料对比研究了尖毛虫属这3个乱序基因的进化。结果表明:（1）Oxytricha sp.（ZJ）与O.nova的小核Actin Ⅰ基因具有相同的乱序模式,区别于其余的尖毛虫属物种;在增加尖毛虫属物种的基础上,对前人推测提出了质疑,我们认为MDS-IES接合处移动现象在乱序MDSs之间并非比非乱序MDSs之间更保守。（2）Oxytricha sp.（ZJ）与O.nova的小核α-TBP基因具有相同乱序模式和相似长度的IESs。（3）Oxytricha sp.（ZJ）的小核DNA pol α基因乱序模式,区别于任一已报道物种,与属内O. trifallax最为相近。基于序列分析,在DNA pol α基因中发现了一例IES转换为MDS的痕迹,以及由此导致原先MDS的丢失。研究发现在编码区内IES向MDS的转变,使得本应删除的序列成为基因组永久保留的一部分。     

长江口地区五种腹毛类纤毛虫的形态学研究

利用活体观察和蛋白银染色技术,对采自长江口泥滩潮间带及河道的五种腹毛类纤毛虫进行了形态学研究。包括对两个国内新记录种仁川巴库虫Bakuella (Bakuella) incheonensis和希斯多利织毛虫Histriculus histrio提供了详细的活体特征和纤毛图式信息,对红色伪角毛虫Pseudokeronopsis rubra、黄色伪角毛虫Pseudokeronopsis flava和冠突伪尾柱虫Pseudourostyla cristata进行了形态学重描述。五种腹毛类纤毛虫的长江口种群与国内外种群均存在不同程度的形态学差异:仁川巴库虫较韩国原始种群体型大,口围带小膜和额前棘毛数较多;希斯多利织毛虫与部分国外种群存在体长差异;红色伪角毛虫与青岛种群相比形态特征基本吻合但个体大小的波动范围较大;黄色伪角毛虫与湛江种群相比额棘毛数目较多;冠突伪尾柱虫较奥地利种群体型较小,与日本种群相比大核较多。该工作丰富了中国腹毛类纤毛虫多样性的认识。     

念珠伪角毛虫小核体功能初步研究

为了研究念珠伪角毛虫的小核是否具有及具有怎样的体功能,采用显微切割手术去小核并建立无小核细胞系。经蛋白银染色鉴定,无小核细胞系群体中大多数细胞的形态结构存在缺陷：口围带的部分小膜缺损或排列紊乱,大核的数目和形态也不正常。这表明,念珠伪角毛虫的小核对于保持口围带结构的完整性以及大核的数目和形态结构的稳定性起着明确的作用。     

甘肃农田四种土壤腹毛类纤毛虫（纤毛门， 旋毛纲，腹毛亚纲）的形态学研究

利用非淹没培养皿法、活体观察法和蛋白银染色法,对采自甘肃临夏县和会宁县农田土壤的4种腹毛类纤毛虫,即柠檬管膜虫(Cyrtohymena citrina)、背触毛拟腹柱虫(Gastrostylides dorsicirratus)、沙栖蠕形尖毛虫(Vermioxytricha arenicola)和半残毛虫属一种(Hemisincirra sp.)进行了研究。结果表明,柠檬管膜虫、背触毛拟腹柱虫和沙栖蠕形尖毛虫为中国纤毛虫分布新记录种。半残毛虫属一种因缺失相关分类学、细胞发生学和分子学信息,此处暂作为未定名种处理。文中对4种纤毛虫的形态学特征进行了详细描述和讨论。     

三种纤毛虫对土壤微生物量和有效氮磷含量的影响

采用土壤培养方法研究了 3种纤毛虫对土壤微生物量及氮磷转化的影响 ,结果表明 ,向土壤接种肾形虫 ( Colopodia sp.)、尖毛虫 ( Oxytricna sp.)和澳毛虫 ( Australothrix sp.) ,特别是澳毛虫 ,显著地降低了土壤微生物碳。说明供试的原生动物与微生物之间存在消长关系。接种澳毛虫显著地降低了土壤有效磷含量 ,而肾形虫和尖毛虫对土壤有效磷含量影响很小 ,仅在培养后期显著地降低了土壤铵态氮含量 ,3种原生动物特别是澳毛虫 ,显著地降低了土壤氮矿化量和硝态氮含量 ,但提高了土壤铵态氮含量 ,说明 3种原生动物抑制了硝化作用 ,而增强了氨化作用。     

苔藓织毛虫的形态和形态发生

应用蛋白银染色和扫描电子显微镜技术研究了苔藓织毛虫的形态和形态发生过程。首次显示了由纤毛、毛基体和基体托架组成的棘毛立体构型,揭示了细胞周期中大核内出现为蛋白银着色的纤维状结构,以及新背触毛原基的产生与裸毛基体的密切关系。     

复合肥对花背蟾蜍蝌蚪红细胞微核及核异常的影响

以花背蟾蜍蝌蚪外周血红细胞微核及核异常作为毒理监测指标,对Gosner 31、42和46期蝌蚪进行观察,研究了复合肥对水生生物的毒性效应.结果表明:用复合肥溶液浓度为1、1.5、2、2.5和3g·L-1进行染毒时,红细胞出现了大、小2种类型的微核,在核异常类型中出现了严重的核分叶和双核类型;在3个被观察的蝌蚪发育期中,42期蝌蚪红细胞微核细胞率和核异常细胞率数值明显低于其他两个发育期;在5个浓度的染毒中,2.5g·L-1浓度组在微核、双核和核分叶与对照组间无显著性差异;花背蟾蜍不同发育时期的蝌蚪,对生境中不良因素的抵抗能力存在着明显的差别,且微核和核异常与复合肥浓度之间没有明显的剂量-效应关系.     

Comparison of Singlet and Doublet Paramecium tetraurelia: DNA Content,Protein Content and the Cell Cycle*

SYNOPSIS Doublet Paramecium tetraurelia contain either a single macronucleus which is substantially larger than that in a singlet cell, or 2 smaller macronuclei. Doublets have approximately twice the DNA content and twice the total protein content of singlets. The cell cycle of doublets is 164% as long as that of singlets, but the relative position of the macronuclear DNA synthesis period within the cell cycle is the same as in singlets. The DNA content of doublets is regulated so that differences in the number of macronuclei do not produce corresponding changes in DNA content; bimacronucleate doublets have only 27% more DNA than unimacronucleate doublets.     

Studies on the Macronuclei of Doublet Paramecium tetraurelia: Distribution of Macronuclei and DNA at Fission*

SYNOPSIS. Doublet Paramecium tetraurelia would be expected to contain 2 macronuclei if their nuclear complement were strictly analogous to that of singlets. However, most doublets are unimacronucleate. It is shown in this study that dimacronucleate cells are present only in young clones. Unimacronucleate cells arise either through abnormalities in the determination and distribution of macronuclear anlagen during the first cell cycle after conjugation, or from dimacronucleate cells through abnormal division and segregation of macronuclei during the fission process. When a change in the number of macronuclei occurs through abnormalities in the division and segregation of daughter macronuclei, the daughter cells produced typically have DNA contents more similar than those expected from either random segregation of daughter macronuclei, or from the normal segregation pattern in ciliates in which changes in the number of macronuclei in progeny cells do not occur. This suggests that part of the regulation process of macronuclear DNA content in Paramecium may occur through control of the segregation pattern of daughter macronuclei.     

棘尾虫无小核镜像骈体的获得及其生殖行为的实验观察

用在分裂期进行显微手术的方法和在陈旧培养中诱导的方法,从贻贝棘尾虫天然无小核系S_(10)中获得了无小核镜像骈体。对骈体的各种生殖行为进行了观察,并用孚尔根和改进的黑色素染色技术揭示了它们的细胞学细节。发现在本文描述的无小核骈体和过去报告的有小核骈体之间,在二分裂和生理再生上没有显著区别。无小核骈体和正常单体S_7之间的接合命运则是多样化的。在630个这样的接合对中,有257对发生了真正的接合并形成接合后体;有153对接合不久包囊化;在其余的220对中,一个接合体吸收了另一个,变成营养型的单体和有小核的骈体。前者占总吸收者的96.8％,后者只占3.2％。对接合和吸收两种情况的细胞学事件进行了详细观察。无小核骈体中的两组细胞质和双套大核能吸引其配偶中的小核,形成自己新的双套大核胚基及小核胚基,在本文的讨论中受到充分注意。     

FURTHER OBSERVATIONS ON THE FINE STRUCTURE OF THE MACRONUCLEUS IN TOKOPHRYA INFUSIONUM

In a previous paper (8) an organized structure was described in the macronuclei of certain old organisms of Tokophrya infusionum. It was found that the same honeycomb structure appears in great abundance in the macronuclei of overfed organisms. This permitted a better three-dimensional reconstruction of the described structure. Since the defined structure may be experimentally induced, it offers an opportunity for further more detailed studies as to its nature and meaning.     

瘦尾虫的小核在无性繁殖周期中对细胞形态结构的影响

通过显微切割建立瘦尾虫无小核细胞系,并与原细胞系及切割后再生的有小核细胞系进行对照观察。结果表明,无小核细胞的形态结构,尤其是口器出现高比率的畸形。在生长静止期,无小核细胞系群体中约23 % (36/155)的细胞完全失去了波动膜。与此同时,这些细胞的口围带也显出异常。一些无小核细胞的大核也出现异常,有的细胞仅含有1枚大核,有的则含有4枚,而不是通常的2枚;还有少数细胞的大核在非细胞分裂期进行分裂。上述结果提示,在无性繁殖周期中,瘦尾虫的小核对于维持正常的细胞形态结构、尤其是保持胞口结构的稳定性和大核数的恒定起着重要的作用[动物学报52 (2) : 383 -388 , 2006]。     

Isolation of a Tetrahymena thermophila strain which induced metaphase I meiotic arrest: new pathway of abortive conjugation

A hypodiploid strain of Tetrahymena thermophila has been obtained that shows arrest at the stage of condensed nuclei, corresponding to metaphase I of normal conjugants and induced arrest at meiotic metaphase I (i.e. at the stage of condensed, bivalent chromosomes) in its wt partner mate. The metaphase I arrested conjugants retained their old macronuclei and most of them underwent cell fusion, instead of separation of exconjugants. The doublets were viable and cortically integrated. When the arrest inducing strain was crossed to the haploid tester strain, the haploid micronuclei were arrested in the meiotic metaphase I as the diploid ones had been; the monovalent, chromosomes were condensed, the arms of sister chromatids were not separated, and they were not segregated. Separation of the arms of sister chromatids and disjunction of bivalent chromosomes were not prerequisite for the formation of microtubular spindles in those cells that were arrested in meiotic metaphase I. After re-feeding, the doublet cells resumed cell divisions, segregating two macronuclei and micronuclei at random. One macronucleus was derived from the arrest inducing strain and the other from the tester strain. Heterokaryon strains with macronuclei derived from the parental arrest inducing strain and with the micronucleus derived from the parental wt tester strain were obtained. Surprisingly, these heterokaryons did not induce meiotic arrest. Thus, the arrest in the melotic metaphase I was induced by the micronucleus and not by the macronucleus of the arrest inducing strain.     

A new AluI satellite DNA in the root-knot nematode Meloidogyne fallax: relationships with satellites from the sympatric species M. hapla and M. chitwoodi

A highly abundant satellite DNA comprising 20% of the Meloidogyne fallax (Nematoda, Tylenchida) genome was cloned and sequenced. The satellite monomer is 173 bp long and has a high A + T content of 72.3%, with frequent runs of A's and T's. The sequence variability of the monomers is 2.7%, mainly due to random distribution of single-point mutations. A search for evidence of internal repeated subunits in the monomer sequence revealed a 6-bp motif (AAATTT) for which five degenerated repeats, differing by just a single base pair, could be identified. Pairwise comparison of the M. fallax satellite with those from the sympatric species Meloidogyne chitwoodi and Meloidogyne hapla revealed a high sequence similarity (68.39%) with one satellite DNA subfamily in M. chitwoodi, which indicated an unexpected close relationship between them. Given the high copy number and the extreme sequence homogeneity among monomeric units, it may be assumed that the satellite DNA of M. fallax could have evolved through some recent and extensive amplification burst in the nematode genome. In this case, its relatively short life would not yet have allowed the accumulation of random mutations in independent amplified repeats. Considering the morphological resemblance between the two species and their ability to produce interspecific fertile hybrids under controlled conditions, these results indicate that M. fallax may share a common ancestor with M. chitwoodi, from which it could have diverged recently. All these data suggest that M. fallax could be the result of a recent speciation process and show that Meloidogyne satellite DNAs may be of interest to resolve phylogenetic relationships among closely related species from this genus.      

DIFFERENTIATION OF THE CHLOROPLAST OF ANTHOCEROS

It has been demonstrated that the following changes accompany differentiation in the plastid of the liverwort Anthoceros eckloni. The inner membrane of the plastid folds to give rise to small vesicles which grow and may fuse to form thylakoids, i.e. flat bags. The thylakoids may "pair" to produce the doublets (thick membranes) of the grana. The doublets may be produced also by the invagination of a thylakoid. In both cases, the doublets are produced only where outside-to-outside contact of thylakoid membranes occurs, which supports the thesis that the thylakoid membranes are polarized. The thylakoids also fold outwards, anastomose, and may fuse. This results in a complicated membrane system, for which an interpretation becomes very difficult. The starch is produced in the matrix, and the pyrenoid bodies are interpreted as specialized regions of the matrix. Younger plastids have grana, but the mature plastid has so many doublets that distinct grana cannot be recognized. This interpretation of the changes which occur during the differentiation of the plastid differs radically from those of Menke (1961) and Manton (1962) who studied this same genus, but is compatible with findings in algae and angiosperms.     

Characterization of six Leuconostoc fallax bacteriophages isolated from an industrial sauerkraut fermentation

Six bacteriophages active against Leuconostoc fallax strains were isolated from industrial sauerkraut fermentation brines. These phages were characterized as to host range, morphology, structural proteins, and genome fingerprint. They were exclusively lytic against the species L. fallax and had different host ranges among the strains of this species tested. Morphologically, three of the phages were assigned to the family Siphoviridae, and the three others were assigned to the family Myovidae: Major capsid proteins detected by electrophoresis were distinct for each of the two morphotypes. Restriction fragment length polymorphism analysis and randomly amplified polymorphic DNA fingerprinting showed that all six phages were genetically distinct. These results revealed for the first time the existence of bacteriophages that are active against L. fallax and confirmed the presence and diversity of bacteriophages in a sauerkraut fermentation. Since a variety of L. fallax strains have been shown to be present in sauerkraut fermentation, bacteriophages active against L. fallax are likely to contribute to the microbial ecology of sauerkraut fermentation and could be responsible for some of the variability observed in this type of fermentation.