鳞毛蕨科3种植物配子体发育的研究

利用光学显微镜详细观察鳞毛蕨科(Dryopteridaceae)3属3种植物,即耳蕨属的棕鳞耳蕨(Polystichum polyblepharum)、鳞毛蕨属的黑足鳞毛蕨(Dryopteris fuscipes)和鞭叶蕨属的鞭叶蕨(Cyrtomidictyum lepidocaulon)配子体发育过程,记录配子体各发育阶段的特征。结果表明:棕鳞耳蕨、黑足鳞毛蕨和鞭叶蕨的孢子都为单裂缝,孢子萌发类型为书带蕨型,配子体发育类型为三叉蕨型,性器官为薄囊蕨型,成熟原叶体为对称的心脏形,具毛状体;3属的属间差异包括孢子颜色、孢子纹饰、萌发时间、丝状体长度、片状体形状、毛状体和假根等特征,上述特征在属间存在交叉现象,因此,孢子形态和配子体发育特征不能作为区分耳蕨属、鳞毛蕨属和鞭叶蕨属的形态依据。     

桫椤科三种植物配子体发育的研究

用土壤培养桫椤Alsophila spinulosa(Wall.exHook.)R.M.Tryon、中华桫椤A.costularis Baker和白桫椤Sphaeropteris brunoniana(Hook.)R.M.Tryon的孢子,利用光学显微镜对其配子体发育的各个阶段进行了观察,包括孢子形态及其萌发、原丝体发育特点、片状体和生长点的形成及分化、假根特征、精子器和颈卵器的分化及发育,以及原叶体感染真菌后的显微特征。初步讨论了桫椤科不同属(种)间的配子体发育的系统学意义和有性世代的濒危原因。     

金粉蕨配子体发育及其无配子生殖的研究

采用光学显微镜和半薄切片技术对金粉蕨(Onychium siliculosum(Desv.)C.Chr.)的配子体发育及其无配子生殖进行了研究。金粉蕨孢子黄褐色,具三裂缝,孢子萌发时先产生假根,然后长出原叶体细胞,萌发类型为书带蕨型。原叶体细胞经丝状体、片状体发育为原叶体,发育类型为水蕨型。金粉蕨配子体分枝发达,成熟配子体呈丛状。多次培养表明金粉蕨只能产生精子器,不能产生颈卵器,为专性无配子生殖。半薄切片观察表明配子体生长点下方的细胞经分裂产生一群小型细胞,由它们发育为无配子生殖胚。小型细胞先分化出无性胚芽,再分化出胚根。胚芽与胚根,胚与配子体之间由分化出的导管连接。     

波叶仙鹤藓的孢子培养及发育生物学研究

首次成功地进行了人工条件下波叶仙鹤藓(Atrichum undulatum(Hedw.)P.Beauv)单细胞孢子的培养,并详细研究、拍摄了其植物体(配子体)形态发育的全过程。孢子无后熟或体眠期,3-4天即可萌发,萌发率可达95％以上;培养10天左右绿丝体系统、轴丝体系统及假根开始分化,约40天开始出现芽体原基,50天芽体陆续形成,80天左右植物体完成营养生长,开始性器发育。初生假根发育不良,基原细胞不分裂,由绿丝体产生轴丝体的方式复杂,轴丝体细胞分裂面有球形和斜向、横向三种,芽原基分化时,首先在轴丝体上产生光合能力很强的细胞群,其基部产生芽原基,芽原基的分化成功率仅有50％左右,芽体形成后,原丝体系统多枯萎或特化为假根。并就波叶仙鹤藓的形态发育、生理生态、生殖和进化等方面也进行了理论探讨。     

骨碎补科两种植物配子体发育的研究

首次报道了骨碎补科(Davalliaceae)的大叶骨碎补(Davallia formosana Hayata)和阴石蕨[Humata repens (L.f.) Diels]配子体发育过程.两者孢子均为单裂缝,椭圆形,不具周壁,书带蕨型(Vittaria-type)萌发;成熟原叶体心形,边缘及背腹面均分布有毛状体;精子器和颈卵器类型为同型孢子薄囊蕨类所具有的一般类型.总结了大叶骨碎补(D.formosana Hayata)和阴石蕨[H.repens (L.f.) Diels]的配子体发育在孢子大小、原叶体边缘细胞形状、精子器和颈卵器形态及毛状体产生时间等方面的异同,初步讨论了这两个种配子体的抗逆性、营养繁殖、假根功能及其可能的系统学意义.     

实蕨和刺蕨（实蕨科）配子体发育的研究

首次在光学显微镜下详细观察比较了实蕨科(Bolbitidaceae)的实蕨(Bolbitis hainanensis Ching et C.H.)和刺蕨[Egenolfia appendiculata (Willd.) J.Sm.]的配子体连续的发育过程.实蕨和刺蕨孢子均为单裂缝,播种5 d 后孢子萌发,萌发类型为书带蕨型(Vittaria-type),播种30～35 d 后发育为片状体.幼原叶体不对称,成熟原叶体具毛状体,呈心形.播种 55 d 后开始有性器出现.通过重点比较丝状体、原叶体、假根及性器等方面的异同,讨论了Holttum和秦仁昌对该科的系统划分,并且进一步支持了秦仁昌的观点.     

野雉尾金粉蕨配子体发育及其系统学意义

野雉尾金粉蕨为中国蕨科金粉蕨属植物,而金粉蕨属的系统位置一直存在争议。该研究用原生境腐殖土和改良克诺普氏(Knop’s)营养液对野雉尾金粉蕨的孢子进行培养,培养条件为温度25℃、光照强度2 500 lx、光照12 h/d,在光学显微镜下观察记录其孢子萌发和配子体发育过程。结果表明:野雉尾金粉蕨的孢子为黄褐色,四面体型,三裂缝,赤道面观为扇形,具周壁,外壁表面具网状纹饰。孢子培养7 d后开始萌发,孢子萌发类型为书带蕨型(Vittaria-type)。孢子萌发后,配子体原始细胞经多次横向分裂形成3~9个细胞的丝状体,丝状体细胞呈圆筒形,壁薄,侧壁向外鼓起,含有颗粒较大且数量较多的叶绿体。15 d左右发育为片状体,片状体多为匙状。25 d左右形成幼原叶体,幼原叶体不对称,配子体发育类型为水蕨型(Ceratopteris-type)。在原叶体发育过程中分枝丝状体非常发达,配子体呈丛状生长,整个发育过程没有毛状体产生。野雉尾金粉蕨的假根为单细胞管状,偶有分支,内无叶绿体。45 d左右精子器开始出现,精子器顶面观近圆形,侧面观为近椭圆形或短柱状。精子器成熟时,盖细胞裂开,精子逸出。60 d左右颈卵器出现,颈卵器比较大,基部略大于顶部,侧面观呈烟囱状,顶面观为铜钱状,颈部由四列细胞构成。90 d左右发育出肉眼可见的幼孢子体。从研究结果看,其配子体发育的特征与凤尾蕨科(Pteridaceae)凤尾蕨属(Pteris L.)相似,支持金粉蕨属归于凤尾蕨科的观点。该研究结果为野雉尾金粉蕨系统学研究提供了配子体发育方面的证据。     

石灰岩地区蕨类植物低头贯众的配子体发育研究

采用原生境腐殖土对低头贯众[Cyrtomium nephrolepioides (Christ) Cop.]孢子进行培养,在显微镜下详细观察了其配子体发育的全过程.结果表明:低头贯众孢子深棕褐色,二面体形,极面观椭圆形,孢子萌发不具假根或为书带蕨型(Vittaria-type);丝状体由3～8个细胞构成;片状体上偏斜具毛状体,宽达10个细胞;原叶体的发育为叉蕨型(Asplenium-type).成熟的原叶体为对称心形,具乳头状毛状体,假根分叉,根尖膨大,精子器壁由3细胞构成和颈卵器弯曲等表现出进化的特征,但假根含有叶绿体又表现了原始的系统学特征.研究认为孢子萌发较慢,假根尖膨大和毛状体等似乎是该种配子体对石灰岩土壤特殊生境的适应对策.     

东方荚果蕨配子体发育的研究

报道在光学显微镜下,观察到的东方荚果蕨配子体发育的过程。培养5 d左右孢子萌发;丝状体单列细胞;片状体发育方式为槲蕨型(D rynaria type);成熟雄配子体近心形,成熟雌配子体呈阔圆心形;毛状体由1~3个细胞组成;精子器发育方式为水龙骨科型(Polypod iaceae type);配子体中部及基部细胞内存在哑铃型叶绿体;在近原丝体细胞中,可观察到极不规则的叶绿体;假根单细胞,丝状。     

亮毛蕨的配子体发育特征及其系统学意义

采用原生境土培养方法对亮毛蕨(Acystopteris japonica)进行培养,并对其配子体发育过程进行了观察。结果表明:亮毛蕨植物的孢子二面体型,极面观为椭圆形,赤道面观肾形,单裂缝,无周壁,外壁具棒状纹饰,孢子萌发为书带蕨型(Vittaria-type),原叶体发育为铁线蕨型(Adiantum-type)。丝状体4～7个细胞,片状体小楔状,仅3～5个细胞宽,成熟原叶体心形,裸露,初生假根具叶绿体,颈卵器较短;配子体发育较迅速,假根具分枝和膨大,精子器囊由3个壁细胞构成,颈卵器略弯曲。本研究首次观察到亮毛蕨的颈卵器壁细胞内含有球形颗粒,但其功能不详。亮毛蕨的配子体发育过程兼有原始和进化的特点。在适宜的光照和温度下,土壤培养亮毛蕨原叶体的成苗率达95%以上,移栽成活率达89%以上。     

Control of sporulation in the filamentous cyanobacteriumAnabaena torulosa

In the cyanobacteriumAnabaena torulosa, sporulation occurred even during the logarithmic growth phase. Sporulation was initiated by differentiation of the vegetative cell on one side, adjoining the heterocyst followed by differentiation of the vegetative cell on the other side. Subsequently, spores were differentiated alternately on either side to form spore strings. The sequence of sporulation supports the previous notion that a gradient of spore maturation exists in cyanobacteria and also indicates that the gradient is manifested unequally on either side of heterocysts. Sporulation was absent or negligible in a minerally enriched medium but ocurred readily in a minimal medium. The extent of sporulation was inversely related to phosphate concentration. Sporulation was enhanced at higher temperature. Incandescent light, but not fluorescent light, greatly stimulated sporulation suggesting possible involvement of red light in spore differentiation. Addition of filtrate, from 5 to 8 day old cultures, to freshly inoculatedA. torulosa greatly enhanced sporulation indicating the influence of extracellular products in spore formation.     

Dictyopyrones, novel alpha-pyronoids isolated from Dictyostelium spp., promote stalk cell differentiation in Dictyostelium discoideum

Dictyopyrones A and B (DpnA and B), whose function(s) is not known, were isolated from fruiting bodies of Dictyostelium discoideum. In the present study, to assess their function(s), we examined the effects of Dpns on in vitro cell differentiation in D. discoideum monolayer cultures with cAMP. Dpns at 1-20 microM promoted stalk cell formation to some extent in the wild-type strain V12M2. Although Dpns by themselves could hardly induce stalk cell formation in a differentiation-inducing factor (DIF)-deficient strain HM44, both of them dose-dependently promoted DIF-1-dependent stalk cell formation in the strain. In the sporogenous strain HM18, Dpns at 1-20 microM suppressed spore formation and promoted stalk cell formation in a dose-dependent manner. Analogs of Dpns were less effective in affecting cell differentiation in both HM44 and HM18 cells, indicating that the activity of Dpns should be chemical structure specific. It was also shown that DpnA at 2-20 microM dose-dependently suppressed spore formation induced with 8-bromo cAMP and promoted stalk cell formation in V12M2 cells. Interestingly, it was shown by the use of RT-PCR that DpnA at 10 microM slightly promoted both prespore- and prestalk-specific gene expressions in an early phase of V12M2 and HM18 in vitro differentiation. The present results suggest that Dpns may have functions (1) to promote both prespore and prestalk cell differentiation in an early stage of development and (2) to suppress spore formation and promote stalk cell formation in a later stage of development in D. discoideum.     

26种丛藓科（Pottiaceae）植物孢子形态及其系统学意义

通过光学显微镜（LM）和扫描电镜（SEM）观察了26种国产丛藓科（Pottiaceae）植物孢子的形态特征,从孢子形态学角度佐证了这些类群在系统演化中的密切关系。结果表明：这26种植物的孢子在形态、近极面形状及孢壁纹饰等方面有较大的相似性,但在孢子大小和表面纹饰的细微结构及其分布等方面又有区别,显示了同科不同属、种间遗传的相似性及遗传分化。     

DIF signalling and cell fate

The DIFs are a family of secreted chlorinated molecules that control cell fate during development of Dictyostelium cells in culture and probably during normal development too. They induce stalk cell differentiation and suppress spore cell formation. The biosynthetic and inactivation pathways of DIF-1 (the major bioactivity) have been worked out. DIF-1 is probably synthesised in prespore cells and inactivated in prestalk cells, by dechlorination. Thus, each cell type tends to alter DIF-1 level so as to favour differentiation of the other cell type. This relationship leads to a model for cell-type proportioning during normal development.     

Inhibitors of intracellular cyclic AMP accumulation affect differentiation of sporogenous mutants of Dictyostelium discoideum

Abstract Sporogenous mutants of Dictyostelium discoideum strain V12M2 were used to determine whether the intracellular levels of cyclic AMP or other second messengers regulate differentiation. Increasing external concentrations of cyclic AMP promoted spore formation. Caffeine and progesterone, which lower intracellular cyclic AMP levels by different mechanisms, blocked spore formation and favored stalk cell formation. In contrast, differentiation of both spore and stalk cells occurred normally in the presence of agents that disrupt calcium/calmodulin or protein kinase C-based second messenger systems. The data are in accord with the view that (1) intracellular cyclic AMP is essential for terminal differentiation of both cell types, and (2) higher levels are required for formation of spores than for stalk cells.     

Involvement of the TRAP-1 homologue, Dd-TRAP1, in spore differentiation during Dictyostelium development

Tumor necrosis factor receptor-associated protein 1 (TRAP1) is a member of the molecular chaperone HSP90 (90-kDa heat shock protein) family. We have previously demonstrated that Dictyostelium discoideum TRAP1 (Dd-TRAP1) synthesized at the vegetative growth phase is retained during the whole course of D. discoideum development, and that at the multicellular slug stage, it is located in prespore-specific vacuoles (PSVs) of prespore cells as well as in the cell membrane and mitochondria. Thereupon, we examined the function of Dd-TRAP1 in prepore and spore differentiation, using Dd-TRAP1-knockdown cells (TRAP1-RNAi cells) produced by the RNA interference method. As was expected, Dd-TRAP1 contained in the PSV was found to be exocytosed during sporulation to constitute the outer-most layer of the spore cell wall. In the TRAP1-RNAi cells, PSV formation and therefore prespore differentiation were significantly impaired, particularly under a heat stress condition. Although the TRAP1-RNAi cells formed apparently normal-shaped spores with a cellulosic wall, the spores were less resistant to heat and detergent treatments, as compared with those of parental MB35 cells derived from Ax-2 cells. These findings strongly suggest that Dd-TRAP1 may be closely involved in late development including spore differentiation, as well as in early development as realized by its induction of prestarvation response.     

Germination and Outgrowth of Single Spores of Saccharomyces cerevisiae Viewed by Scanning Electron and Phase-Contrast Microscopy

Single spores of Saccharomyces cerevisiae were examined during germination and outgrowth by scanning electron and phase-contrast microscopy. Also determined were changes in cell weight and light absorbance, trehalose utilization, and synthesis of protein and KOH-soluble carbohydrates. These studies reveal that development of the vegetative cell from a spore follows a definite sequence of events involving dramatic physical and chemical modifications. These changes are: initial rapid loss in cellular absorbance followed later by an abrupt gain in absorbance; reduction in cell weight and a subsequent progressive increase; modification of the spore surface with concomitant diminution in refractility; elongation of the cell and augmentation of surface irregularities; rapid decline in trehalose content of the cell accompanied by extensive formation of KOH-soluble carbohydrates; and bud formation.     

Folic acid stimulation of the Gα4 G protein-mediated signal transduction pathway inhibits anterior prestalk cell development in Dictyostelium

In Dictyostelium discoideum, several G proteins are known to mediate the transduction of signals that direct chemotactic movement and regulate developmental morphogenesis. The G protein alpha subunit encoded by the Galpha4 gene has been previously shown to be required for chemotactic responses to folic acid, proper developmental morphogenesis, and spore production. In this study, cells overexpressing the wild type Galpha4 gene, due to high copy gene dosage (Galpha4HC), were found to be defective in the ability to form the anterior prestalk cell region, express prespore- and prestalk-cell specific genes, and undergo spore formation. In chimeric organisms, Galpha4HC prespore cell-specific gene expression and spore production were rescued by the presence of wild-type cells, indicating that prespore cell development in Galpha4HC cells is limited by the absence of an intercellular signal. Transplanted wild-type tips were sufficient to rescue Galpha4HC prespore cell development, suggesting that the rescuing signal originates from the anterior prestalk cells. However, the deficiencies in prestalk-specific gene expression were not rescued in the chimeric organisms. Furthermore, Galpha4HC cells were localized to the prespore region of these chimeric organisms and completely excluded from the anterior prestalk region, suggesting that the Galpha4 subunit functions cell-autonomously to prevent anterior prestalk cell development. The presence of exogenous folic acid during vegetative growth and development delayed anterior prestalk cell development in wild-type but not galpha4 null mutant aggregates, indicating that folic acid can inhibit cell-type-specific differentiation by stimulation of the Galpha4-mediated signal transduction pathway. The results of this study suggest that Galpha4-mediated signals can regulate cell-type-specific differentiation by promoting prespore cell development and inhibiting anterior prestalk-cell development.