THE ULTRASTRUCTURE OF MITOSIS AND CYTOKINESIS IN THE SARCINOID CHLOROKYBUS ATMOPHYTICUS (CHLOROPHYTA,CHAROPHYCEAE) REVEALED BY RAPID FREEZE FIXATION AND FREEZE SUBSTITUTION1

Mitosis and cytokinesis in vegetative cells of the sarcinoid green alga Chlorokybus atmophyticus Geitler were examined with rapid freeze fixation, freeze substitution, and transmission electron microscopy. The taxonomic placement of C. atmophyticus in the class Charophyceae sensu Stewart and Mattox is corroborated by some mitotic and cytokinetic features including development of a microtubular sheath around the prophase nucleus, the almost constant chromosome to pole distance during anaphase, telophase nuclei widely separated by a persistent interzonal spindle, and centripetal plasma membrane invagination. Features, previously unknown in the Charophyceae, include the specific position of the peroxisome lying between the nucleus and adjacent cell wall during interphase and mitosis, the extensive array of microtubules radiating from the centrioles located at the presumptive poles at prophase, involvement of coated vesicles in the furrowing process, and occurrence of transversely aligned cleavage microtubules. Placement of Chlorokybus in the order Klebsormidiales is proposed.     

ULTRASTRUCTURE OF MITOSIS AND CYTOKINESIS IN KLEBSORMIDIUM MUCOSUM NOV. COMB., FORMERLY ULOTHRIX VERRUCOSA (CHLOROPHYTA)1

A transmission electron microscopy study of dividing cells of Ulothrix verrucosa Lokhorst has provided clear evidence that this species differs in many respects from other Ulothrix Kützing species. These differences include the presence of a microtubular sheath around the prophase nucleus, the complete disintegration of the nuclear envelope coinciding with the proliferation of extranuclear microtubules into the prometaphase nucleus and the intrusion of vacuoles into the interzonal spindle region in between the widely separated telophase nuclei. This necessitates the transfer of Ulothrix verrucosa to the charophycean genus Klebsormidium Silva, Mattox and Blackwell. The new combination Klebsormidium mucosum is proposed. On account of its mitotic pattern, this species can be placed in the (charophycean) evolutionary line towards the higher plants. However, because of its cytokinesis (annular centripetal ingrowth of the plasmalemma) this species probably should be considered as a blind offshoot of this line. It is emphasized that furrowing green algae with a persistent interzonal spindle at telophase (including the presently studied alga) often show an ill-defined cytokinetic microtubular system.     

MITOSIS AND CYTOKINESIS IN SESSILE SPORANGIUM OF TRENTEPOHLIA AUREA (CHLOROPHYCEAE)1

An ultrastructure study of mitosis and cytokinesis in the sessile sporangium of Trentepohlia aurea (L.) Mart, was made to clarify the phylogenetic position of the alga. Mitosis was closed and centric at late anphase with cytokinesis involving the production of cleavage membranes by dictyasames between the numerous, well-separated daughter nuclei. Neither phycoplast nor phragmoplast microtubules were observed during cytokinesis. The lack of phycoplast microtubules and the presence of multilayered structures in flagellated cells suggest Trentepohlia is phylogenetically related to those green algae thought to have given rise to the land plants. The primitive type of mitosis and the lack of microbodies suggest that the ancestors of Trentepohlia may have branched off from this line relatively early.     

CELL DIVISION IN THE FILAMENTOUS PLEURASTRUM AND ITS COMPARISON WITH THE UNICELLULAR PLATYMONAS (CHLOROPHYCEAE)1

At prophase in Pleurastrum, extranuclear spindle microtubules develop from the region of centrioles, which lie lateral to the nucleus midway between the future sites of the metaphase spindle poles. The microtubules then move laterally to overarch the nucleus and finally become incorporated into the spindle. The centrioles do not migrate and therefore lie in the same plane as the chromosomes at metaphase. At telophase, 2, more different systems of microtubules develop from the vicinity of the centrioles—a phycoplast and extensive arrays of microtubules that ensheath the daughter nuclei. Cell division in the filamentous Pleurastrum is compared to that in the green flagellate, Platymonas. The similarities between cell division in the 2 algae are interpreted as evidence: (i) that rhizoplasts (which in Platymonas resemble myofibrils) are somehow homologous to microtubules; and, (ii) that cell division in Pleurastrum differs from cell division in other examined filamentous chlorophycean genera because Pleurastrum has an independent evolutionary origin from a monad with Platymonas-like characteristics.     

FINE STRUCTURAL STUDIES ON THE INTERPHASE AND DIVIDING APICAL CELLS OF SPHACELARIA TRIBULOIDES (PHAEOPHYTA)1

The apical cells of Sphacelaria tribuloides Menegh. are larger than other thallus cells, contain more organelles and appear polarized. Their tip portion, where they grow, contains a well developed Golgi apparatus, abundant endoplasmic reticulum (ER) membranes, mitochondria, chloroplasts and a large number of small vacuoles. It seems likely that a continuous flow of membranous material from the ER membranes to the dictyosomes and from the latter to the plasmalemma of the extending tip portion takes place. In contrast, the basal pole possesses fewer organelles and is occupied mainly by large-sized, sometimes central vacuoles. The apical cells undergo two distinct types of highly asymmetrical differential divisions giving rise to cells of the thallus and hair initials. During the early stages of mitosis the nuclear envelope remains intact, except for fenestrated poles. Microtubules pass through the fenestrae into the nucleoplasm. During meta-phase, a typical chromosome plate is organized. The sites of attachment of spindle microtubules to the chromosomes are structurally different from the rest of the chromosomes. At late anaphase, the nuclear envelope breaks down completely. During telophase, a new membrane encloses the chromosomes which are decondensed and the nucleoli are reorganized. Cytokinesis proceeds long after mitosis at a stage in which the nuclei have increased in size and have moved farther apart. A membranous furrow develops centripetally, without the participation of microtubules. However, microtubules traverse the thin cytoplasmic strands which, in both interphase and cytokinetic cells, meander among the vacuoles of the basal pole of the cell and the internuclear space. Dictyosomes appear to be involved in the subsequent wall deposition.     

CELL DIVISION AND COLONY FORMATION IN THE GREEN ALGA COELASTRUM (CHLOROCOCCALES)1

Multinucleate cells of Coelastrum undergo precisely directed cytokinesis, guided by phycoplast microtubules, to form a number of uninucleate daughter cells which subsequently adhere to form characteristically patterned aggregates. As there is no movement of the daughter cells relative to one another before their adhesion, the disposition of cells in daughter colonies reflects the pattern of cytokinesis of parent cells. Centrioles lie at the poles of the mitotic nuclei which are partially enclosed by a perinuclear envelope of endoplasmic reticulum. The centrioles disappear at the time of cytokinesis of the parental cell and apparently reform de novo once the daughter cells have acquired a cell wall following their adhesion. The trilaminar layer of cell wall, often termed the pectic layer, does not stain with ruthenium red and resists acetolysis suggesting that it contains sporopollenin rather than pectin.     

MITOSIS AND CYTOKINESIS IN ASTEROMONAS GRACILIS, A WALL-LESS GREEN MONAD1

Asteromonas gracilis Artari remains motile throughout cell division. Basal bodies separate and replicate at prophase. They are located lateral to the poles of the closed metaphase spindle. Kinetochores appear at late metaphase. Chromosomes move to the poles and extensions of the nuclear envelope develop into the pyrenoid at anaphase. The interzonal spindle disintegrates at telophase and a diffuse phycoplast is present. Cytokinesis proceeds rapidly from the anterior region of the cell. Newly formed daughter cells have four narrow-banded rootlets and both distal and proximal fibers connect the basal bodies. Features of cell division in Asteromonas are compared to those in other algae, particularly Dunaliella and Chlamydomonas.     

红豆杉细胞悬浮培养生产紫杉醇研究进展

介绍了近年来由红豆杉细胞培养生产紫杉醇领域取得的进展,其中特别介绍了由紫杉醇生物合成途径及其代谢酶类的研究发展而来的用基因工程方法改良细胞系,为提高紫杉醇产量而采取添加前体物质,诱导子,抑制子等方面的研究。     

紫杉醇及其衍生物的研究概况与发展趋势

本文分别从生物资源,药理学,化学合成和组织培养方面对红豆杉植物中紫杉醇及其衍生物的研究工作进行了综述,从中可以了解紫杉醇及其衍生物的研究概况与发展趋势。     

PHRAGMOPLASTS IN CYTOKINESIS OF CEPHALEUROS PARASITICUS (CHLOROPHYTA) VEGETATIVE GELLS 1

Cytokinesis in apical cells of actively growing cultures of Cephaleuros parasiticus Karsten sporangiate thalli was examined with transmission electron microscopy. A massive, interzonal cytokinetic microtubule spindle is anchored at its poles to the medial surfaces of the daughter nuclei at telophase. Later, the daughter nuclei are widely separated and no longer associated with the interzonal spindle; however, the spindle retains its shape and becomes a distinct phragmoplast with an array of vesicles, presumably derived from dictyosomes, aligned in the division plane. Fusion of the vesicles gives rise to a thin cell plate. Some bundles of microtubules in the spindle appear to mark the sites of plasmodesmata formation, but no endoptasmic reticulum is directly involved in plasmodesmata formation. No infurrowing or phycoplast array of microtubules is involved in the cytokinesis. The relationship, if any. between the metaphase-anaphase mitotic microtubule system and the interzonal cytokinetic spindle has not been determined. Cephaleuros parasiticus isone of only four green algae now known to contain a higher plant-like phragmoplast and cytokinetic process. The observations reported can be interpreted as very strong evidence for a phylogenetic affinity between the Trentepohliaceae and the Charophyceae, but consideration of ulvophycean features of the Trentepohliaceae such as motile cell ultrastructure and life histories precludes unequivocal assignment of the family to either the Charophyceae or Ulvophyceae.     

MITOSIS IN THE OCTAFLAGELLATE PRASINOPHYTE,PYRAMIMONAS AMYLIFERA (CHLOROPHYTA)1

Cell division is described in the octaflagellate prasinophyte Pyramimonas amylifera Conrad and is compared in related genera. Basal bodies replicate at preprophase and move toward the poles. Cells remain motile throughout division. The nuclear envelope disperses and chromosomes begin to condense at prophase. Pairs of multilayered kinetochores are evident on the chromosomes of the metaphase plate. Spindle microtubules extending from the region of the basal bodies and rhizoplasts attach to the kinetochores or extend from pole to pole. Numerous vesicles and ribosomes have entered the nuclear region and the incipient cleavage furrow invaginates. The chromosomes move toward the poles at anaphase leaving a broad interzonal spindle between the two chromosomal plates. The nuclear envelope reforms first around the chromatin on the side adjacent to the spindle poles and later on the interzonal side. The cleavage furrow progresses into the interzonal spindle at telophase. By late telophase the nucleoli have reformed and the chromosomes have decondensed. The interzonal spindle has not been observed late in telophase. As the cleavage furrow nears completion the cells begin to twist and contort, ultimately separating the two cells.     

稀土化合物对悬浮培养红豆杉细胞紫杉醇生产和释放的影响

采用稀土化合物硝酸镧、硫酸铈铵、硝酸亚铈处理悬浮培养的红豆杉细胞,对八个非外泌型细胞株进行了研究。它们的紫杉醇产量在０．０５￣１５．４４ｍｇ／Ｌ之间,释放率在０％￣２７％之间不同的细胞株对不同稀土化合物的响应是不同的。除ＴＮ、ＮＦ细胞株紫杉醇产量有所下降外,其他细胞株均表现紫杉醇的产量不同程序的提高;Ｅ２细胞株对稀土元素的促渗透作用不敏感。硝酸镧、硫酸铈铵、硝酸亚铈三种稀土化合物中,硝酸亚铈对Ｄ４     

The effect of cytochalasin J on kinetochore structure in PtK1 cells is mitotic cycle dependent

Mitotic PtK1 cells were arrested in mitosis with nocodazole to determine the effect of cytochalasin J (CJ) on kinetochore structure in arrested and nocodazole-released cells. In previous studies it was shown that CJ had a more pronounced effect on alteration of kinetochore structure and spindle microtubule (MT) architecture when applied during prophase or prometaphase. In this study, mitotic cells were treated at preanaphase for 10 min with 1 microg/ml nocodazole, or in 1 microg/ml nocodazole and 10 microg/ml CJ to allow for the advancement of the 'mitotic clock'. Thus it can be determined if either changes in the timing of mitosis, the maturation of the kinetochore, and/or the lack of MT connection to the kinetochore affects the ability of CJ to detach or alter the attachment of chromosomes to the developing spindle. Preanaphase cells treated with 1 microg/ml nocodazole for 10 min and released into 10 microg/ml CJ showed significant changes in MT organization and kinetochore structure. MTs nucleated at the centrosome are fragmented and kinetochore structure was significantly altered showing only two laminae with few MTs inserted into this structure. Preanaphase cells treated with 1 microg/ml nocodazole and 10 microg/ml CJ for 10 min and released into 10 microg/ml CJ showed similar, but more pronounced, effects on kinetochore structure and spindle MT organization. We interpret these results to suggest that CJ treatment has a greater effect on MT attachment and kinetochore structure in nocodazole pre-treated cells, where the kinetochore structure is mature and the mitotic cycle has been advanced.     

紫杉醇在氧化铝层析过程中的稳定性研究

本文比杉醇在氧化铝层析分离紫杉烷过程中的稳定性进行了实验分析。结果表明,紫杉醇在柱中的仪时间、流动相中甲醇浓度、介质的含水量以及操作温度是影响稳定性的主要因素。当停留时间控制在４５ｍｉｎ以内,氧化是质经过脱水处理,降低甲醇浓度及温度控制在２５℃以内时,可减少紫杉醇在氧化铝层析过程中的降解。     

紫杉醇替代资源研究进展

紫杉醇是近30年来从植物中筛选出的首选抗癌药物,已被美国的FDA批准治疗晚期卵巢癌和转移性乳腺癌。由于资源奇缺已出现严重供给危机。本文综述了近3年来紫杉醇替代资源研究的新进展,主要包括组织培养、遗传转化、化学合成和紫杉霉等方面的新成就。     

抗癌药物紫杉醇的全合成—Holton合成紫杉醇路线的剖析

从全合成战略与路线,关键反应,保护基应用和立体化学方面对Holton的线性全合成紫杉醇路线进行剖析归纳。     

Dynamic distribution of Ser-10 phosphorylated histone H3 in cytoplasm of MCF-7 and CHO cells during mitosis

The dynamic distribution of phosphorylated Histone H3 on Serl 0 (phospho-H3) in cells was investigated to determine its function during mitosis. Human breast adenocarcinoma cells MCF-7, and Chinese hamster cells CHO were analyzed by indirect immunofluorescence staining with an antibody against phospho-H3. We found that the phosphorylation begins at early prophase, and spreads throughout the chromosomes at late prophase. At metaphase, most of the phospho-H3 aggregates at the end of the condensed entity of chromosomes at equatorial plate. During anaphase and telophase,the fluorescent signal of phospho-H3 is detached from chromosomes into cytoplasm. At early anaphase, phospho-H3 shows ladder bands between two sets of separated chromosome, and forms “sandwich-like structure” when the chromosomes condensed. With the cleavage progressing, the “ladders” of the histone contract into a bigger bright dot. Then the histone aggregates and some of compacted microtubules in the midbody region are composed into a “bar-like”complex to separate daughter cells. The daughter cells seal their plasma membrane along with the ends of the “bar”,inside which locates microtubules and modified histones, to finish the cytokinesis and keep the “bar complex” out of the cells. The specific distribution and kinetics of phospho-H3 in cytoplasm suggest that the modified histones may take part in the formation of midbody and play a crucial role in cytokinesis.     

抑制剂促进紫杉醇生物合成的初步研究

研究了５－甲基－ＤＬ－色氨酸（５－ＭＴ）,丙二酸钠等１１种代谢抑制剂对红豆杉悬浮培养细胞生长和紫杉醇生物合成速率的影响,筛选取５－ＭＴ等３种对紫杉醇生物合成有促进作用的抑制剂,抑制剂在稳定期添加剂红豆杉细胞生长无强烈的抑制作用。更有利于紫杉醇的生物合成,各抑制剂与前体和真菌诱导协同作用提高增产效果,增产大小依次为５－ＭＴ,马来酸和丙二酸钠,其中５０ｍｇ·Ｌ＾－１５－ＭＴ使紫杉醇含量提高近１０倍,１