四膜虫S1有性生殖的电镜观察Ⅱ.接合区的形态与配子核的交换

四膜虫接合膜上的小孔是两接合体细胞质相连的通道。配子核形成后,按合膜由于增生而出现装有细胞器等的囊状折叠,并可脱离下来而落入另一细胞中,这可能是胞质交流的又一途径。配子核的交换不是从膜上原有小孔通过的,而是在溶酶体等作用下、使膜破裂,由核后方的微管推动进入对侧细胞中。 本文记述了四膜虫S1有性生殖过程中接合区的形态和配子核的交换。     

上海四膜虫和两株嗜热四膜虫的rRNA基因ITS-1序列及分子系统关系

测定并比较了自接型的上海四膜虫（Tetrahymena shanghaienisis)和两株接合型的嗜热四膜虫（T.thermopddhilaⅡ和T.thermophilaⅥ）的ITS-1序列,以多态嗽叭虫（Stentor polymorphrus)为外来群,利用最大简约法和邻接法构建了它们的系统发育树。分析指出：三者中,T.shanghaienisis较早地从祖先种中分化出来;自接型可能是一种较接合型原始的生殖方式。     

四膜虫S1株——上海四膜虫,新种

四膜虫S1是一株自接型四膜虫,根据形态特征应属梨形四膜虫复合种。本种除克隆内接合外,和其它十二种四膜虫均不接合。本种的异柠檬酸脱氢酶,四唑氧化酶,谷氨酸脱氢酶和乙酸酯酶的同功酶谱和其它十二种四膜虫相比,差异明显。此外,金亦石等(1987)所提供的S1和其它十种四膜虫的rDNA分子的四种限制性内切酶图谱也说明,S1株与除T.australis以外的其它九种的酶切图谱是显著不同的。据此,作者认为本种应是梨形四膜虫复合种中一新种,并定名为上海四膜虫。     

四膜虫S1营养生长和有性过程中cAMP的含量变化

用放射免疫法测定了四膜虫在群体生长各阶段和在有性过程中cAMP的含量变化。在群体生长的延迟期(Lag phase)之末,即将进入对数期时,cAMP含量出现一个峰值。随着对数期的进展,cAMP含量陆续下降,在对数期之末,cAMP含量降至最低值。群体进入静止期后,细胞内cAMP含量略有回升。 在饥饿诱导接台的过程中,细胞内cAMP含量在开始饥饿的很短时间内即迅速降至一个最低点,并且在整个接合过程中一直保持较低的水平。接合抑制剂伴刀豆球蛋白A(CoaA)和精胺(spermine)等在抑制接合的同时,也阻止饥饿细胞cAMP含量的迅速下降,使之不能达到在正制情况下接合出现时所达到的低水平。 在饥俄细胞的培养液中检出了较高量的cAMP。这可能有助于说明何以饥饿细胞的cAMP含量能够在很短时间内迅速下降。至于排出的cAMP对四膜虫的接合有无促进或其他作用,有特继续研究。     

上海四膜虫接合生殖期间皮层细胞骨架蛋白的研究

本文采用生化抽提,结合电泳及显微技术,对上海四膜虫（Ｔｅｔｒａｈｙｍｅｎａｓｈａｎｇｈａｉｅｎｓｉｓ）Ｓ１皮层细胞骨架（ｃｏｒｔｉｃａｌｃｙｔｏｓｋｅｌｅｔｏｎ）的蛋白组份,及其在接合生殖期间的变化进行了一系列的研究,初次探索了Ｓ１株上海四膜虫在接合生殖中皮细胞骨架的蛋白组份及含量,并分析了它们与间期,接合分开时期同类蛋白相互间的差异,发现在接合生殖时期７６－８８ｋＤ蛋白有突出的表现,而９０ｋＤ和     

携带编码苜蓿根瘤菌结瘤基因的根癌农杆菌和大肠埃希氏菌,可在苜蓿根上形成类根瘤

将编码苜蓿根瘤菌(Rhizobium meliloti)结瘤基因(nod)的质粒,通过接合作用转移到根癌农杆菌(Agrobacterium tumefaciens)和大肠埃希氏杆菌(Eschrichia coli)里,分别形成转移接合体。这两种转移接合体都能诱导苜蓿根毛卷曲,并形成类根瘤。在由根癌农杆菌转移接合体形成的类根瘤里,可看到典型的根瘤分     

棘尾虫接合生殖期间大核功能的研究

为了澄清棘尾虫接合期间大核对小核发育和皮层形态发生的作用,完成了大核摘除,放线菌素D处理,H~3-尿嘧啶核苷标记等实验。摘核实验证明,一个接合体的大核可以通过细胞质桥支持另一除去大核的接合体发育。即使保留接合对大核总数的1/8,这种支持作用仍然存在。还发现来自大核的支持物作用于形态发生更易于作用于小核发育,并对两个接合体的形态发生作用相等,对两个接合体小核发育的作用不等。摘除四分之三大核的实验证明,小核发育和皮层更新在接合后15小时内都不能脱离对残留大核的依赖。放线菌素D处理实验证明,接合后RNA的合成需积累到8.5小时,才可满足核与皮层发育的需要。H~3-尿嘧啶核苷标记实验也支持接合后前9小时内RNA都在持续合成的结论。本文对摘核实验和放线菌素D处理实验结果的差别、以及本文结果与前人结果的差别都做了讨论。     

棘尾虫有性生殖的发动和诱导自配的研究

本文观察了棘尾虫的接合反应。按照接合早期接合状态的变化,将接合反应分成10个时期,并将不同时期的接合对人工拆开,令其继续发育,然后计算每一时期中由营养虫转变为有性过程的虫体的百分比。结果表明,有性过程的发动是在Ⅳ期。一旦发动即不能逆转。将Ⅳ期被拆开的虫体横切成相等的前后切块,发现一些前切块由营养状态转变为有性状态,发生了自配,而所有后切块保持营养状态,发生了再生。这一现象意味着有性过程的发动不是整个细胞的一种突然改变,看来是一种从前向后的信息传递过程。用上述人工方法诱导出棘尾虫的自配,核器发生的观察表明,自配体中的桉行动和自配后体中核胚基的发育,所有这些核的事件都与接合生殖的相似,只不过没有两个配偶之间迁移原核交换罢了。对伴随着自配发生皮层重组也做了观察,自配的形态发生也与接合中的一样。     

棘尾虫接合生殖期间核对形态发生的影响

本文完成了四项实验,即1.促成棘尾虫无小核系与有小核系接合;2.诱导无小核系自系接合;3.在正常接合对上摘除全部大核或一个接合体的大核及小核;4.在正常接合后体上对老大核碎块和新大核胚基进行损伤实验。以改进过的黑色素法显示这些实验的结果,发现第一次接合形态发生和小核的有性过程都主要是由大核支持的,小核在第一次形态发生中也起重要作用。这些核产物可从一个有核接合体穿过细胞质桥去支持另一无核接合体的发育。本文还发现接合第二次形态发生由新大核胚基控制。损伤染色体和多线染色体时期的大核胚基,第二次形态发生消失或不正带。越过第二次形态发生的大核胚基显著增强耐受损伤的能力,这些结果符合Prescott等(1973)对棘尾虫大核遗传装置的设想。     

四膜虫功能基因组数据库增量更新2016:生活史和减数分裂转录组及磷酸化蛋白组资源建设

原生动物嗜热四膜虫是一种优良单细胞真核模式生物,以其作为研究对象在基础生物学领域的研究已经取得了一系列突破性的成果。2006年,其大核基因组测序完成并发表,标志着四膜虫的研究进入了功能基因组时代。2013年基于基因芯片、基因网络和转录组数据,我们构建了四膜虫功能基因组数据库,其目前已成为模式生物嗜热四膜虫研究的两个重要数据库之一。在过去几年里,随着对四膜虫功能基因组学研究的深入,相关组学数据也实现了一定积累,对这些数据的整合也非常迫切。基于此,我们对四膜虫功能基因组数据库进行了增量更新。更新内容主要包括三个方面:(1)四膜虫生活史不同时期转录组数据;(2)四膜虫接合生殖减数分裂过程转录组数据;(3)磷酸化蛋白组数据。此次增量更新进一步提升和完善了四膜虫功能基因组数据库的内容和功能,对以四膜虫为对象的相关研究工作具有重要作用。     

TIME LAPSE ANALYSES OF SEXUAL REPRODUCTION IN CLOSTERIUM EHRENBERGII (CONJUGATOPHYCEAE)1

The process of sexual differentiation was studied using heterothallic clones of Closterium ehrenbergii Meneghini. The first visible sign of sexual reproduction was agglutination of two or more cells in a group and this was followed by gametangiogenic division and conjugation of gametangial cells. Movements of gametangial cells were carefully studied. Gametangial cells occasionally participated again in gametangiogenesis instead of proceeding directly to the formation of conjugation papilla. The whole process of sexual differentiation from vegetative cell to zygospore was considered to be basically similar in both of the two closely related mating groups, A and B, of C. ehrenbergii. Nevertheless, there were some differences between the two groups in patterns of the sexual differentiation. In Group A, vegetative cell division was completely suppressed by mixing the two complementary mating type clones together into the same medium with high light illumination. This suppression was not caused by the nitrogen depletion in the medium, but by the presence of cells of opposite mating type. In Group B, vegetative cell division and sexual reproduction occurred side by side repeatedly for several days.     

Macronuclear Regeneration and Cell Division in Paramecium caudatum

SYNOPSIS. In Paramecium caudatum , occurrence of macronuclear regeneration is closely related to the time of feeding after conjugation. Macronuclear regeneration is induced with a high frequency when conjugating pairs are transferred into fresh culture medium. Feeding immediately after conjugation induces early cell division and 3 or more fissions occur without macronuclear division because of the inability of the macronuclear anlagen to divide. In the cells lacking normal macronuclear anlagen, old macronuclear fragments undergo regeneration and form vegetative macronuclei.     

Mating Types, Breeding System, Conjuàtion and Nuclear Phenomena in the Marine Ciliate Euplotes cristatus Kahl from the Gulf of Naples

SYNOPSIS. The marine ciliate Euplotes cristatus Kahl (Ciliophora, Hypotrichida), collected off Capri, Gulf of Naples, is described in detail. From populations, 6 different mating types, representing 1 variety or syngen, have been isolated. The breeding relations revealed a multiple mating type system characteristic of other members of the Hypotrichida that have been investigated. Presumably a 7th mating type was found which does not mate with any of the others. Although this may belong to another syngen, it could represent a mating type which has not yet reached sexual maturity or 1 which may be in a period of decline. Animals of different mating type do not mate immediately after being mixed but usually 3 or more hours later. An agglutination reaction involving many specimens is absent. Instead, 2 ciliates engage in a “pairing play” before joining firmly in conjugation. Well-fed or actively feeding and dividing ciliates do not mate; mating occurs only after the food becomes gradually depleted or when the food supply is sharply cut off. All mating types appear to be extremely stable. Neither selfing pairs (intraclonal conjugation) nor autogamy have been observed within any clonal culture during the several years under investigation. Cell-free filtrates from 1 mating type do not elicit mating or induce conjugation with specimens of a different mating type. The general pattern of nuclear events in conjugation and exconjugant reorganization is as follows: 1 preliminary division, 3 pregamic (prezygotic) divisions, fertilization, and generally 1 or occasionally 2 postzygotic divisions. The fate of micronuclear products may be determined by their size and location. Those which are larger and close to the cell membranes of the joined conjugants persist and/or divide. Those which are smaller are carried by cyclosis toward the center of each ciliate and degenerate. The degenerating macronucleus of each conjugant becomes segmented in a more or less uniform manner resulting in 4 subspherical masses. Two become localized in the anterior end of a conjugant and 2 in the posterior end. Those in the posterior end are always the first to degenerate completely and disappear. In nuclear reorganization of the exconjugant, fusion of the macronuclear anlage with parts of the old macronucleus does not occur.     

Light Conditions for Sexual Reproduction in Heterothallic Strains of Closterium

Sexual reproduction in heterothallic strains of Closterium peracerosum-strigostan-littorale,KAS-4-29 (mating type minus) and KAS-4-30 (mating type plus),depended on a light intensity higher than 3,000 lux. Two kindsof cell division occurred in the mating medium: 1) one kindoccurred at low light intensity (less than 1,000 lux) and wasnot linked to conjugation, and 2) a sexual one occurred at highlight intensity (higher than 3,000 lux) and was linked to conjugation.3-(4-Chlorophenyl)-l,l-dimethylurea at 1 µM added at thebeginning of the mating culture completely inhibited conjugationbut not cell division. Chloramphenicol at 20 to 40 µg/mladded at the beginning of the culture inhibited conjugationcompletely and cell division partly. The drugs added after 12h of the mating culture did not inhibit the division processes.Varying light intensity after 12 h of culture did not affectsexual activity. The photosynthetic activity was highest inthe cells at the beginning of the mating culture, and then markedlydecreased. The data indicate that the early period of the matingprocess depends strictly on the light conditions. (Received March 12, 1982; Accepted November 30, 1982)     

Flagellar Adherence of Crithidia fasciculata Cells in Culture1

In early log phase cultures of several of the drug-resistant mutants of Crithidia fasciculata that we have previously obtained, a high percentage of cells attach in pairs at the base of the flagellum. This process, which we have termed “flagellar adherence,” lasts for several hours in some cases and occasionally involves changes in cell morphology. The attachment occurs optimally in gently agitated cultures. Flagellar adherent pairs can be disassociated by vigorous agitation; the pairs reappear in the culture within one to three h after disassociation. These paired forms can be clearly distinguished from the normal cell division forms. Clones of flagellar adherent-competent mutant strains are uniformly able to form these pairs in culture. A low percentage of flagellar adherent forms can be induced in wild type cells by glucose starvation.     

Surface changes in differentiating Friend erythroleukemic cells in culture.

The sensitivity to agglutination by several plant lectins has been studied during the induced erythroid differentiation of Friend erythroleukemic cells in culture. In addition, the number of lectin receptors on the cell has been measured. It is shown that early during the differentiation, there is an increase in agglutinability while the receptor density remains constant. In the later phase of the differentiation process, the cells lose their sensitivity to agglutination while the receptor number and density increases. These changes were not observed on nonerythroid mastocytoma culture cells. Two nondifferentiating variants of the FL cells were shown to have altered sensitivities to agglutination by ConA.     

Disturbance of the determination of germinal and somatic nuclei by heat shock in Paramecium caudatum

During conjugation of Paramecium caudatum, nuclear determination occurs soon after the third postzygotic division: one of the four anterior nuclei becomes the micronucleus and the remaining three degenerate, while four posterior nuclei differentiate into macronuclear anlagen. Macronuclear differentiation is supposed to be dependent on a cytoplasmic differentiation factor. In this study, postzygotic cells were subjected to heat shock for 30 min and nuclear changes were observed by staining with carbol fuchsin solution. When heat shock was initiated during the period from metaphase to telophase of the third postzygotic division, cells showed an excess of macronuclear anlagen and were typically amicronucleate. Abnormal nuclear localization around the end of the third (last) postzygotic division may explain the origin of these kinds of cells. A similar phenomenon appeared after treatment with actinomycin D or emetine. Since heat shock did not inhibit macronuclear differentiation but destroyed the formation of micronuclei, some factor(s) probably plays an essential role in nuclear determination, especially in the protection of the micronuclei.     

Effect of organics on sulfur-utilizing autotrophic denitrification under mixotrophic conditions

Thiol groups were introduced to dermal bovine collagen (DBC) by the reaction with gamma-thiobutyrolactone. Thiolated DBC reacted with 2-pyridyl disulfide group introduced to lysozyme to form DBC-lysozyme conjugate through disulfide bridge. The enzymatic activity of freshly prepared conjugate was almost unchanged during ten consecutive runs over one month. The DBC-lysozyme conjugate showed the maximum activity at pH 6.3, on the contrary, that of native lysozyme was pH 9.0. Thermal stability of lysozyme was enhanced by the conjugation with DBC. The present results showed that the conjugation using thiolated collagen could be one of the useful alternative approaches to modify collagen with bioactive molecules.     

The relationship of gonadal development to the life cycles of the paired species of lamprey, Lampetra fluviatilis (L.) and Lampetru planeri (Bloch)

Gonadal developnient has been studied throughout the larval period of Lumpetru fluviatitis and comparisons have been made with conditions in the larvae of the closely relatcd non-parasitic species, L. planeri . The two forms differ in the time of onset or mitotic division in germ cells and in the period when oogenesis is initiated. These phases begin in L. pluneri when the ammocoetes are one year old and in L. fluviatilis at the end of the second year. The course of sex differentiation appears to be sinlilar in both forms. While in the early stages, the oocytes of L. planeri are larger than those of L. fluviatilis. at comparable ages, by the end of the third year they are sinlifar in size in both spccics. It is suggested that in L. planeri , during the extension of the larval period, gonad growth is retarded and during this phase the ammocoete accumulates body reserves which are used to meet the metabolic requirements of the rapidly developing gonads after metamorphosis. The differences observed in early gonadogenesis are believed to account for the reduced fecundity of the L. planeri ammococtc and their adaptive significance is discussed.     

Cytology of an Indian Race of Blepharisma undulans (Stein)

SYNOPSIS. The Indian race of Blepharisma undulans described in this paper measures 150 μ in length. The macronucleus consists of 5–7 nodes, all of equal size. During binary fission, condensation of macronucleus is followed by its elongation and a thinning of the middle region which breaks with the division of the animal. It later attains the typical vegetative form.
During conjugation 7 or 8 micronuclei pass through the first pregamic division, 5 to 7 through the second pregamic division and one product of the second division takes part in the third division. The rest degenerate. At the same time, the macronucleus also starts degenerating. After the synkaryon has divided twice, the conjugating pairs separate. Of the 4 products, 3 become macronuclear anlagen and one, micronuclear anlage.
The micronuclei divide asynchronously both during binary fission and during conjugation. There is apparently considerable diversity in the structure and behaviour of the macronucleus and micronuclei in the different races of Blepharisma undulans.