宽礁膜的生活史

在国内首次观察描述了宽礁膜(Monostroma latissimum Wittrock)生活史各阶段的形态结构特征和发育变化,并对其单性生殖进行了周年培养和观察。大型的雌雄异体的膜状配子体成熟后,放散双鞭毛的配子,小型的球状孢子体成熟后放散四鞭毛的游孢子,其生活史是单倍体的配子体与二倍体的孢子体互相交替的异形世代交替。另外,在生活史中还发现合子和孢子囊的二分裂增殖方式,一年生配子体和二年生孢子囊等现象。未接合的雌、雄配子固着后变成球状单细胞体,室内培养后发育成成熟的单性孢子囊,并放散四鞭毛的游孢子,这些游孢子固着后,可直接发育成膜状的配子体,单性生殖也可以用来进行人工育苗。     

观察蕨类生活史的好材料——铁线蕨

铁线蕨分布广泛,容易培养,是观察蕨类生活史的好材料。将经过低温处理（4℃）的铁线蕨成熟孢子接种到培养基上,在光照培养箱中培养45 d左右,孢子萌发后长成配子体,培养60 d左右,颈卵器内的受精卵萌发长出幼孢子体。将铁线蕨孢子接种到花盆内的土壤上,在室内培养2～3个月长成配子体,培养4～5个月长出幼孢子体。将培养基上的幼孢子体移栽到花盆土壤中可在室内长期培养,四季常青,孢子囊陆续分化、成熟。     

关于"世代交替"的一些疑惑

世代交替是指一些生物的生活史中有双倍体的孢子体世代与单倍体的配子体世代交替出现的现象, 由于涉及孢子、配子、孢子体、配子体等概念,学生常常容易混淆,并产生一些疑惑,现分析如下:     

单倍体在海带遗传研究中的应用

几年来从海带的遗传实验中发现海带单倍体(配子体和配子)是遗传研究的良好材料。科研工作从分离单个海带配子体开始,由此得到了雌配子体和雄配子体的无性生殖系、雌性孢子体和雄性孢子体。这些不同来源的配子体和孢子体在性质上都是“纯系”。配子体的无性生殖系寿命可以很长,它们都能保持原来的性别,而且年年可以由此得到孢子体。由于雌性孢子体能产生大量的雌配子体,完成自己的生活史,用雌配子体来分析海带孢子体性状的遗传情况,或用来研究杂种优势和突变,都很适宜。     

石莼——绿藻繁殖的实验材料

近年,我们发现生长在沿海潮间带的石莼(Ulva lactuca L.)可以用作观察绿藻繁殖的材料。现将实验方法介绍如下: 石莼的生活史具有同型世代交替现象,春季是其繁殖盛期。此时,藻体内的营养细胞能陆续转化成单细胞的生殖器官。孢子体营无性生殖,孢子囊开始在发生叶状体的上部边缘,以后逐渐向藻体内方、下方扩展。成熟的孢子囊内可形成8—16个游动孢子。配子体营有性生     

扇蕨配子体发育与濒危机制的探讨

采用改良Knop’s固体培养基、原生境腐殖质土和红壤分别培养扇蕨（Neocheiropteris palmatopedate）孢子,光学显微镜及解剖镜下观察记录其孢子萌发及配子体发育过程,比较了3种培养方法对其配子体发育和有性繁殖的影响,并在此基础上探讨了扇蕨的濒危原因。结果表明：成熟孢子黄褐色,赤道面观为豆形,极面观椭圆形,单裂缝。孢子萌发类型为书带蕨型,原叶体发育为槲蕨型。成熟原叶体呈心脏形。毛状体在原叶体阶段出现。有性生殖周期长及配子体发育成幼孢子体的百分率低是扇蕨在配子体世代的主要濒危原因。此外,红壤固有的理化性质导致扇蕨配子体发育极其缓慢、精子器和颈卵器发生的时间间隔过长使其不能受精产生孢子体。原生植被遭受破坏引起的林下腐殖质土消失、红壤裸露,加剧了扇蕨的濒危。     

雄配子体选择的遗传分化效应及其在植物改良中的应用

在植物世代交替的生活史中,配子体是产生配子和具有单倍数染色体的植物体,并且有自己的遗传表达信息.在配子产生的过程中以及配子结合之前,适者生存的法则对配子发挥着选择作用,只有最适应外界环境条件的配子才能通过竞争性受精并产生合子.雄配子体选择是影响植物遗传分化、演变和遗传多样性的重要因素,被认为是生物进化的有力动力.此外,由于植物基因组中约有2/3的基因表达交错发生在配子体阶段和孢子体阶段,因此雄配子体选择的结果会影响到下一代孢子体的表现型.在育种实践中,利用雄配子体选择对植物进行遗传改良,具有提高选择机率和缩短育种年限等优点.本文主要概述雄配子体选择与孢子体表型的关刎系、遗传分化效应及其在植物遗传改良中的应用,以期为雄配子选择相关研究提供参考     

4种蕨类植物的孢子繁殖研究

蕨类植物的资源开发越来越受到重视,但目前的孢子繁殖技术还不够成熟,已成为其市场扩大的制约因素。本研究对4种有代表性的资源型蕨类植物乌毛蕨(Blechnum orientale)、荚果蕨(Matteuccia struthiopteris)、假鞭叶铁线蕨(Adiantum malesianum)和水蕨(Ceratopteris thalictroides)进行了孢子繁殖研究,详细观察并记录了繁殖过程中配子体发育各阶段出现的特征和时间。其中以水蕨的发育周期最短,从播种到形成幼孢子体约33天;假鞭叶铁线蕨发育周期最长,约73天;乌毛蕨和荚果蕨的发育周期介于二者之间。结合过去已发表的相关观察资料,对4种蕨类的孢子萌发、原叶体、性器官以及幼孢子体形成等四个生长关键点所出现时间的培养条件进行了分析,结果表明:(1)光照时间越长,孢子萌发天数越短;(2)光照强度越大,原叶体发育天数越长;(3)配子体发育速率在20~25℃范围内与温度无显著相关性。本研究可为进一步探究蕨类植物适宜的繁殖技术体系和开发蕨类资源提供指导。     

渐尖毛蕨和齿牙毛蕨的配子体发育研究

采用混合土培养渐尖毛蕨和齿牙毛蕨的孢子,显微镜下观察记录了它们的孢子萌发及配子体发育过程.结果表明:两者的孢子均为深褐色,极面观为椭圆形,赤道面观为半圆形,单裂缝;渐尖毛蕨的孢子萌发所需时间较渐尖毛蕨短,但两者萌发类型均为书带蕨型;丝状体阶段均发达;原叶体边缘均可产生少量毛状体;成熟原叶体均呈心脏形;由原叶体发育成幼孢...     

利用涂片法观察蕨类植物孢子发生发育

孢子是联系蕨类植物孢子体世代和配子体世代的重要环节。利用涂片法观察瓶尔小草孢子发生发育具有操作简便、不需要化学试剂、观察效果直观等特点,适用于孢子囊较大的种类。涂片法相对来说更适合在《植物学实验》教学中使用,以丰富实验教学内容,培养学生实验技能和学习兴趣。     

Parthenogenesis is rare in the reproduction of a sexual field population of the isogamous brown alga Scytosiphon (Scytosiphonaceae,Ectocarpales)

Parthenogenetic development of unfused gametes is commonly observed in laboratory cultures among various brown algal taxa. There is, however, little information on the contribution of parthenogenesis to the reproduction of field populations. In this study, we investigated whether parthenogenesis is present in a sexual population of the isogamous brown alga Scytosiphon with a 1:1 sex ratio. In culture, both female and male gametes showed higher mortality and slower development compared to zygotes. More than 90% of surviving partheno‐germlings formed parthenosporophytes irrespective of the culture conditions tested. Therefore, if parthenogenesis occurs in the field, most unfused gametes are expected to form parthenosporophytes. Contrary to this expectation, parthenosporophytes were rare in the field population. We collected 126 sporophytic thalli and isolated and cultured a unilocular sporangium from each of them. We confirmed that cultures of 120 unilocular sporangia produced both female and male gametophytes by the observation of zygotes or amplification of PCR‐based sex markers indicating that these sporangia originated from zygotic sporophytes. Only females were detected in cultures from two sporangia and only males from four sporangia suggesting that these sporangia originated from parthenosporophytes. In the Scytosiphon population, although parthenogenesis is observable in culture, our results demonstrate that the contribution of parthenogenesis to reproduction is small (≤4.8%) compared to sexual reproduction. Unfused gametes may not survive to form mature parthenosporophytes in significant numbers in the field partly due to their higher mortality and slower development compared from zygotes.     

The life history ofAcrochaete wittrockii (Ulvellaceae,Chlorophyta)

Acrochaete wittrockii (Wille) Nielsen is a heteromorphic diplohaplont. The haplophase consists of isomorphic, dioecious filamentous epiphytes on brown algae. Several generations follow each other by triflagellate zoospores from spring to early summer. By late summer and throughout autumn, quadriflagellate zoopores are produced by the epiphytic thalli; they give rise to male and female gametophytes of a globular, pseudoparenchymatic appearance in culture. The gametophytes produce anisogamic biflagellate gametes which, after gametic union, develop into diploid unicellular sporophytes. After 6–7 days, the sporophyte produces triflagellate zoospores, repeating the life history when germinating on brown algal hosts. Alternatively, triflagellate zoospores which settle on the bottom of petri dishes, develop into unicellular, autonomous sporangial plants. Their triflagellate spores repeat the epiphytic stage on brown algal hosts, or the sporangial plant cycle on non-living substrate, respectively.     

THE DEVELOPMENT AND REPRODUCTIVE MORPHOLOGY OF HALOSACCION AMERICANUM I. K. LEE (RHODOPHYTA,PALMARIALES)1

Halosaccion americanum, a member of Palmariaceae, was grown in culture from spores and the life history was critically examined by the use of scanning EM and light microscopy. A mature tetrasporangium of H. americanum produces four spores that germinate to form two male and two female gametophytes. The male gametophytes grow to maturity in approximately eight months and macroscopically resemble the tetrasporophyte. Following the first division of the tetraspore, the two-celled female gametophyte consists of a vegetative cell and a carpogonium with trichogynes. Fertilization is accomplished by spermatia from male plants of the preceding generation, as male plants of the same season are immature. Spermatia are formed in a continuous layer over the surface of the mature male gametophytes and, when released, are entrained in long strands of mucous. Spermatia adhere to and fuse with trichogynes and, nuclear fusions presumably follow. The carposporophyte is absent; the new tetrasporophyte develops directly from the fertilized carpogonium. Growth of the sporophyte eventually obliterates the female gametophyte, and development into a mature tetrasporophyte proceeds over a period of approximately eight months. The development of H. americanum, with its extremely abbreviated female gametophyte stage and direct development of the tetrasporophyte from the zygote, indicates that this rhodophyte has the same life history as reported for other members of the Palmariales.     

Morphology and life history of Halopteris filicina (Sphacelariales, Phaeophyceae) from Korea

The vegetative morphology and life history of Halopteris filicina (Grateloup) Kutzing, collected from Korea, were examined in laboratory culture. Field plants attaining 3–5 cm in height were epilithic, tufted, yellowish-brown, and produced numerous erect axes with alternately distichous branches from compact basal discs. They were cultured under a 12:12 h LD photoperiod at 10°-C, 15°C and 20°C to observe the influence of temperature on reproduction. At 10°C plants grew only vegetatively, whereas at 15°C and 20°C they produced unilocular sporangia. Unispores released from sporangia developed into monoecious, anisogamous gametophytes that formed plurilocular female and male gametangia on the same lateral branches. The zygotes, by fusion of female macrogametes and male microgametes, developed into sporophytes bearing unilocular sporangia, whereas the unfused female gametes germinated parthenogenetically. This species was confirmed to have an isomorphic life history, basically similar to the other species of Sphacelariales.     

In vitro fertilization as a tool for investigating sexual reproduction of angiosperms

In vitro fertilization (IVF) of isolated male and female gametes of flowering plants was first accomplished in the last decade. Successful isolation of male and female gametes, and culturing of in vitro zygotes to form new plants, is a prelude to the use of IVF for research into the cellular and molecular control of fertilization in higher plants and its application as a tool in biotechnology. Genes unique to male and female gametes and zygotes of higher plants, although currently incompletely characterized, are expected to permit direct molecular dissection of fertilization. By applying IVF and microculture to zygotes and endosperm obtained by both in vivo and in vitro methods, newly activated fusion products may be observed and manipulated in media where they are directly accessible to the techniques of molecular cell biology. IVF and zygote culture may also offer potential for creating new hybrid plants by fusing isolated gametes from different species to produce unique zygotes and ultimately plants that would be impossible to obtain using typical crossing techniques. Transformation and regeneration frequencies using IVF may also be high enough to avoid the necessity of adding controversial antibiotic and herbicide resistant genes to screen transformed products. This review describes advances using IVF in plant sexual reproduction and discusses its potential in the genetic improvement of flowering plants.     

LIFE HISTORY OF COLPOMENIA SINUOSA (SYCTOSIPHONACEAE,PHAEOPHYCEAE) IN THE AZORES1

Colpomenia sinuosa (Mertens ex Roth) Derbès and Solier (Scytosiphonaceae, Phaeophyceae) is a common species on the rocky intertidal shores of the Azores, where reproductive gametophytes occur throughout the year. Life‐history studies of this species were carried out in culture, and both sexual and asexual reproduction were observed. Anisogamous gametes fused to form zygotes. The zygotes gave rise to a filamentous prostrate sporophyte generation bearing unilocular sporangia, under both short‐day and long‐day conditions at 15 and 22°C, and to both unilocular and plurilocular sporangia, under the lower temperature condition. Unispores developed into gametophytes, and plurispores gave rise to filamentous sporophytes. Asexual reproduction was carried out by unfused female gametes and asexual plurispores produced from the same gametophyte. Unfused gametes developed into filamentous prostrate sporophytes producing unilocular sporangia in both culture conditions, and unispores released from the sporangia gave rise to gametophytes. Asexual plurispores from field gametophytes, under both culture conditions, developed directly into new gametophytes. The species exhibited three types of life history: a heteromorphic, diplohaplontic; a heteromorphic, monophasic (both with alternation between the erect and filamentous prostrate thalli); and a monomorphic, monophasic.     

Gametic behavior in a marine green alga, Monostroma angicava: an effect of phototaxis on mating efficiency

The role of phototactic behavior of gametes was tested experimentally in the slightly anisogamous marine green alga Monostroma angicava Kjellman, and the effect of phototaxis on mating efficiency was discovered. Both male and female gametes showed positive phototaxis in response to a white light source. In contrast, they did not respond to a red light source. Their swimming velocity did not differ between these two illuminating light sources. It was, therefore, suggested that the search ability of the gamete itself might not vary between phototactic and non-phototactic conditions. The number of zygotes formed during the mating process may be expressed as the product of the number of encounters between male and female gametes and the fraction of encounters that result in sexual fusion. In this study, with high densities of male and female gametes mixed in test tubes, almost all minor (fewer in number) gametes fused sexually within 10 min. After dilution of the gamete suspensions by half, mating efficiency in test tubes illuminated by white light from above was higher than that in dark controls. This suggests that male and female gametes gathered at the water surface through their positive phototaxis, thus increasing the rate of encounters. Mating efficiency also decreased if the test tubes were illuminated from above by white light and also shaken. Since negative phototaxis is clearly shown in planozygotes, we suggest that positive phototaxis of male and female gametes in M. angicava is an adaptive trait for increasing the rate of gametic encounters rather than for the dispersal of zygotes as previously reported for zoospores of some marine algae. Received: 12 February 1999 / Revision accepted: 24 May 1999     

The fate of chloroplast DNA during cell fusion, zygote maturation and zygote germination in Chlamydomonas reinhardi as revealed by DAPI staining

Chlamydomonas reinhardi, a haploid isogamous green alga, presents a classic case of uniparental inheritance of chloroplast genes. Since the molecular basis of this phenomenon is poorly understood, an examination of the cytology of the C. reinhardi plastid DNA was made in gametes, newly formed zygotes, maturing zygotes, and at zygote germination.The single plastid per cell of Chlamydomonas contains a small number of DNA aggregates (‘nucleoids’) which can be seen after staining with DNA-binding fluorochromes. In zygotes formed by pre-stained gametes, the fluorescing nucleoids disappear from the plastid of mating type minus (male) gamete plastids but not from the plastid of mating type plus (female) gamete plastids about 1 h after zygote formation. Subsequently, nucleoids aggregate slowly to a final average of two or three in the single plastid of the mature zygote.Quantitative microspectrofluorimetry indicates that gametes of both mating types have equal amounts of plastid DNA, and that zoospores arising from zygotes have 3.5 × as much as gametes. Assuming degradation of male plastid DNA, there must be a very major synthesis of plastid DNA between zygote formation and zoospore release when zygotes produce the typical 8–16 zoospores. That synthesis appears to occur at germination, where there is a massive increase in plastid DNA and nucleoid number beginning just prior to meiosis. The results support the theory that uniparental inheritance results from degradation of plastid DNA entering the zygote via the male gamete and suggest further studies, using mutants and altered conditions, which might explain how male plastid DNA sometimes survives.     

Analysis of gamete and zygote motility in Allomyces

To study the mechanisms of chemotaxis in eukaryotes, the motility patterns of the gametes and zygotes and the chemotactic responses of the male gametes of the lower eukaryote Allomyces macrogynus were examined. Dark-field microscopy of the male gametes showed a smooth swimming pattern interrupted by very brief ‘jerks’ of the cell body that caused a change in swimming direction. Female gametes had a slower swimming velocity than the males and underwent more jerks or turns which accounted for their sluggish motility. The zygotes swam with the fastest velocity and were observed to have a helical swimming pattern involving a continuous turning of the cell body, a behavior absent from the gametes. Introduction of female gametes that produce the chemoattractant sirenin brought about an immediate change in the behavior of the male gametes. They moved in spirals (or helices) towards the source of the chemoattractant (the female gametes), undergoing only a few jerks to reorient the male cells. When very near the female cells, or in high concentrations of added sirenin, many very short motility tracks were observed that finally resulted in contact between the two gamete types. The results indicate that the poor swimming ability of the female gametes facilitates gamete contact, resulting in as many as 30–40 male gametes clustered on a single female cell. Further, male gamete orientation to the sirenin gradient is caused by the chemoattractant suppressing the jerk motion.