植物雌配子体发育研究进展

高等植物雌配子体的形成涉及孢原细胞和大孢子母细胞的确立与分化、大孢子发生、功能大孢子以及胚囊的形成和发育等多种复杂调控过程。随着当代生物技术及功能基因组学的发展,近年对雌配子体发育的研究已从细胞学描述逐渐过渡到对基因和发育调控分子机理的探索。以拟南芥、水稻和玉米等模式植物为材料进行的相关研究,丰富了人们对于植物雌配子体和其它有性生殖过程遗传调控机理的认识。本文着重阐述了植物雌配子体发生和发育过程,并综述了这一领域最新研究进展。     

植物雄配子体发生和发育的遗传调控

植物雄配子体发生和发育是有性生殖的关键步骤之一,是高等植物通过有性生殖进行世代交替所必需的。近几年来,随着分离和鉴定配子体型突变体技术的发展,雄配子体遗传机制研究取得了很大进展,发现了一些调控雄配子体发生和发育过程中细胞分化和互作的重要基因。本文着重概述和讨论植物雄性生殖细胞和雄配子体发生及其与周边细胞互作遗传机制研究的最新进展。     

Gametogenesis in Plasmodium; the inhibitory effects of anticytoskeletal agents

Gametocytes of Plasmodium yoelii were incubated with colchicine, vinblastine sulphate and cytochalasin B and then induced to undergo gametogenesis. All the drugs inhibited gamete formation in a dose and time dependent manner. Electron microscopy revealed that colchicine and vinblastine sulphate inhibited the polymerisation of cytoplasmic- axonemal- and intranuclear mitotic- microtubules. Cytochalasin B did not inhibit microtubule assembly but blocked spindle pole separation and axoneme distribution into the microgamete. All drugs prevented escape of the gametocyte from the erythrocyte. The mechanism of mitotie spindle action is discussed.     

非交叉配子形成体的连锁图谱构建方法

根据非交叉(achiasmatic)遗传模型,提出采用最大似然法计算遗传交换率的方法,同时开发了构建非交叉生物(F2群体)连锁图谱的计算机软件。通过卡方验检可测性连锁分子标记。对于无交叉生物现象,采用蒙特卡洛模拟技术,对交叉(chiasmatic)和非交叉两个遗传模型遗传交换率的估计值和作图效率进行了比较。模拟结果表明,非交叉模型能提供无偏的估计值,而交叉模型则只有实际值的一半。在所有同等的条件下,基于非交叉模型的作图效率均高于基于交叉模型(无校正)的作图效率。对于非交叉配子形成体,采用基于非交叉模型的交换率计算方法能获得理想的作图效率。     

Effect of Fixed Daylengths on the Photoperiodic Regulation of Gametogenesis in the Sea Urchin Strongylocentrotus purpuratus

Summary

The gametogenic response of the sea urchin Strongylocentrotus purpuratus was compared under treatments of different photoperiod regimes of fixed and seasonal changing daylength. Quantification of gametogenic activity in en- zymatically dissagregated ovaries and in histological sections of testes have shown that the sea urchin is sensitive to both fixed and variable daylengths. After one year at fixed short day (8L:16D) or fixed neutral day (12L:12D) the gonads were ripe and in active gametogenesis, the same as were gonads of animals reared at changing photoperiod and sampled during the short-day phase of the cycle. Under fixed long day (16L:8D) the gonads lacked significant amount of gametes and had the cell constitution found in gonads of animals reared at changing photoperiod and sampled during the long-day phase of the cycle. Measurement of a critical daylength related to the autumnal equinox seems to be part of the mechanism in the photoperiodic regulation of vitellogenic oocyte growth and spermatogenesis in these animals.     

TFIID在配子发生和早期胚胎发育过程中的作用

配子发生以及胚胎早期发育过程受严格且有序的基因表达调控。多种转录因子与靶基因结合,激活基因的时空特异性表达,实现受精卵全能性的获得,完成母型基因组转录调控向合子基因组转录调控的转变以及随后胚胎细胞的分化调节。研究表明,TFIID转录因子家族在这些关键阶段起重要作用,在基因转录调节的起始阶段,TFIID转录因子家族成员作为通用转录因子被招募到靶基因的启动子上,与其他转录因子共同形成转录前起始复合物,起始转录。该文总结了TFIID转录因子的结构、作用方式,以及在配子发生和早期胚胎发育中的调控作用。     

苞叶姜大小孢子发生和雌雄配子体发育

对苞叶姜（Pyrgophyllum yunnanense）大小孢子发生和雌雄配子体发育进行了显微观察,研究结果表明：苞叶姜花药2室,药壁发育属基本型,绒毡层为分泌型。小孢子母细胞经减数分裂形成左右对称型四分体,胞质分裂为连续型,成熟花粉粒为2-细胞型;子房下位,3室,胚珠多数,倒生,具双珠被、厚珠心,中轴胎座。大孢子母细胞经减数分裂形成线型（或T形）四分体,珠孔端3个大孢子退化,合点端的1个发育成功能大孢子,胚囊发育为蓼型。比较苞叶姜、黄花大苞姜（Caulokaempferia coenobialis）和海南三七（Kaem pferia rotunda）的早期胚胎学特征,表明苞叶姜与黄花大苞姜系统学关系较近,与海南三七系统学关系较远。在分类等级上,胚胎学证据支持苞叶姜独立成属。     

Gametogenesis and chromosome number in Postelsia palmaeformis (Laminariales, Phaeophyceae)

Gametophytes of the ‘sea palm’, the kelp Postelsia palmaeformis Ruprecht, produced gametes whether or not chelated iron was supplied in the culture medium, in contrast to the inhibition of gametogenesis seen with the absence of iron in many other kelps. As gametogenesis proceeded, every cell of the gametophytes was converted into a gamete so that the gametophytes did not continue to grow vegetatively. The portion of the life history from spore release through germination, gametophyte growth, gametogenesis, fertilization and growth of the young sporophyte was completed in 9–10 days under laboratory conditions. Chromosome counts showed that sporophytes had a diploid number of 26–34 chromosomes while sporangia and gametophytes had a haploid number of 14–17 chromosomes, indicating a typical haplodiplophasic life history as seen in other Laminariales.     

Primary Production and Calcification by the Soritid Foraminifer Archais angulatus (Fichtel & Moll)*

SYNOPSIS. Symbiosis between Chlamydomonas hedleyi (Lee, Crockett, Hagen & Stone) and Archais angulatus (Fichtel & Moll) was examined during laboratory studies of primary production and light-enhanced calcification. Photosynthesis and calcification are directly proportional to light intensity in the range of 0–200 μEinsteins m^-2 sec^-1. Calcification in the light is directly proportional to photosynthesis and proceeds at rates that are 2–3 times that observed in the dark. The herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), in concentrations of 1–100 μM, completely inhibits photosynthesis and light-enhanced calcification. Calcification of the foraminiferan test is therefore due to the photosynthetic activity of the symbiote. Calcification rates for foraminifers incubated in the dark or with DCMU are not significantly different from the calcification rates obtained for dead foraminifers. Rates of calcification obtained with ⁴⁵Ca are twice that obtained with ¹⁴C.     

Two Classes of Gap Junction Channels Mediate Soma-Germline Interactions Essential for Germline Proliferation and Gametogenesis in Caenorhabditis elegans

In all animals examined, somatic cells of the gonad control multiple biological processes essential for germline development. Gap junction channels, composed of connexins in vertebrates and innexins in invertebrates, permit direct intercellular communication between cells and frequently form between somatic gonadal cells and germ cells. Gap junctions comprise hexameric hemichannels in apposing cells that dock to form channels for the exchange of small molecules. Here we report essential roles for two classes of gap junction channels, composed of five innexin proteins, in supporting the proliferation of germline stem cells and gametogenesis in the nematode Caenorhabditis elegans. Transmission electron microscopy of freeze-fracture replicas and fluorescence microscopy show that gap junctions between somatic cells and germ cells are more extensive than previously appreciated and are found throughout the gonad. One class of gap junctions, composed of INX-8 and INX-9 in the soma and INX-14 and INX-21 in the germ line, is required for the proliferation and differentiation of germline stem cells. Genetic epistasis experiments establish a role for these gap junction channels in germline proliferation independent of the glp-1/Notch pathway. A second class of gap junctions, composed of somatic INX-8 and INX-9 and germline INX-14 and INX-22, is required for the negative regulation of oocyte meiotic maturation. Rescue of gap junction channel formation in the stem cell niche rescues germline proliferation and uncovers a later channel requirement for embryonic viability. This analysis reveals gap junctions as a central organizing feature of many soma–germline interactions in C. elegans.     

Proteases of Melilotus alba mesophyll protoplasts

Proteases from mesophyll protoplasts of Melilotus alba were identified by standard proteolytic assays and separated using different chromatographic techniques. Their characterization also included their subcellular location. Besides the evidence for the multiplicity of the proteolytic enzymes, two protease sets were distinguished endopeptidases, which are exclusively vacuolar, and aminopeptidases, which are widely distributed throughout the cell. Cytosol-located enzymes were tested as substrates of the two sets of proteases, by studying comparatively the time-course changes of enzyme activities during incubation in total protoplast extracts, or in cytosol fractions devoid of vacuolar proteases. The degradation of phosphoenolpyruvate-carboxylase protein, a typical cytosolic enzyme, in the presence of purified amino-and endopeptidases, was also estimated by immunoprecipitation studies. Only the vacuolar endopeptidases are effective in the degradation of cytosolic enzymes. Hydrolytic enzyme activities mostly of vacuolar origin were very stable during incubation in total protoplast extracts. These proteins therefore appear to be particularly resistant to proteolytic attack. The results indicate that, in plants, the effective proteolytic system acting on cytosolic enzymes seems to be vacuole-located, and that the selectivity in protein degradation may be imposed by the susceptibility of the protein being degraded and by its transfer into the vacuoles.Abbreviations Leu-pNA leucine-p-nitroanilide - lys-p-NA lysine-p-nitroanilide - pCMB p-chloromercuribenzoic acid - PEPCase phosphoenolpyruvate carboxylase - PMSF phenylmethylsulfonylfluoride - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Gametogenesis of Chlamydomonas in the dark

When cells of each mating type of Chlamydomonas sp. (Toyonakastrain) were transferred to a nitrogen-free medium and culturedat low temperature (10?C) for 10 to 12 hr, they showed matingactivity even in the dark. The highest activity observed inthe dark, at about 10?C, was similar to that in the light at20?C. Induction in the dark was suppressed by oxygen deficiency.It was also inhibited by addition of 1 to lOmin concentrationsof nutritional nitrogen compounds such as ammonium, nitrateor urea, and of cycloheximide (2 to 20 µg/ml), but notby addition of chloramphenicol, dihydrostreptomycin or spectinomycin.On the other hand, it was slightly promoted by addition of theophylline,aminophylline, cAMP, DB-cAMP, kinetin and abscisic acid. ¹Present address: Department of Animal Virology, The ResearchInstitute for Microbial Diseases, Osaka University, Suita, Osaka565, Japan. (Received June 3, 1976; )     

Glutamine synthetases of green and etiolated leaves ofSinapis alba

Studies on the glutamine synthetases (GS, EC 6.3.1.2) of green (GS₂) and etiolated leaves (GS_et) ofSinapis alba L. (cv. Steinacher) revealed striking similarities between the respective enzyme proteins. The enzymes showed corresponding chromatographic properties, both on dimethylaminoethyl-Sephacel and on hydroxylapatite columns. The purified GS proteins were also identical with regard to the molecular weight of their subunits. Isoelectrofocusing of pure GS_et yielded two distinct polypeptide bands in the pH 5.6 region of the gels. This pattern corresponded to the two strong bands of GS₂. Two charge variants of GS polypeptides could be detected by Western-blot analysis of the soluble protein of green leaves using antibodies against mustard GS₂. In immunoprecipitation experiments, the holoenzymes of GS₂ and GS_et were recognized with identical affinities by this antiserum. We conclude that strong similarities exist between the proteins of the GS enzymes in green and etiolated leaves of mustard. Most probably only one GS form, namely the plastidic enzyme, can be found in the epigeal organs ofSinapis. The polypeptides of the GS₂ subunits showed no differences in the hydrophobicity of the polypeptide chains. Neither glucosyl nor mannosyl residues could be detected. Dedicated to Professor Dr. H. Mohr on the occasion of his 60th birthday     

A Brief Report on the Chromosome Number of Neodiplogaster pinicola and Panagrellus redivivoides (Nematoda: Rhabditida)

Both Neodiplogaster pinicola and Panagrellus redivivoides reproduce amphimictically, with XO type of sex determination. In N. pinicola, primary spermatocytes have six bivalent chromosomes and one univalent; after two meiotic divisions, sperm are produced with either six or seven chromosomes. In primary oocytes, with seven bivalents, meiosis is initiated by entrance of a sperm. After two meiotic divisions, three polar nuclei are produced, and egg and sperm pronuclei fuse. Cleavage begins after the egg is laid. Males have a 2n number of 13 chromosomes; females, 14. In P. redivivoides, primary spermatocytes have four bivalents and one univalent. After two meiotic divisions, spermatids are produced with either four or five well separated chromosomes. In primary oocytes, the first maturation division is initiated after penetration of a sperm; after two meiotic divisions, each egg has five chromosomes. Cleavage begins immediately after fusion of egg and sperm pronuclei, and embryonic development and hatching occur within the uterus. Males have a 2n chromosome number of 9; females, 10.     

Algal Flagellate Symbiosis in the Foraminifer Archaias*†

SYNOPSIS. A symbiotic alga from the foraminifer Archaias angulatus was isolated axenically. Algae from crushed hosts were coccoid and highly vacuolated; division stages within an envelope were common. Biflagellate motile piriform organisms predominated in newly transferred cultures and were gradually replaced by the coccoid, highly vacuolated stage. Incorporation of ¹⁴C label in intact Archaias was greatest for organisms fed and incubated in light. Starved symbiotized organisms incubated in the light incorporated ～30% as much label as fed counterparts, There was no obvious difference in ⁴⁵Ca incorporation between fed and starved organisms. Light significantly enhanced calcification. The Archaias symbiont infected Rosalina leei but not Quinqueloculina spp.