狭眼凤尾蕨配子体发育及其成精子囊素对水蕨配子体发育的影响

对狭眼凤尾蕨（Pteris biaurita）配子体发育特征及其外源成精子囊素对模式植物水蕨（Ceratopteris thalictroides）在黑暗和光照条件下孢子萌发和配子体发育的影响进行了研究。结果表明：（1）狭眼凤尾蕨孢子深褐色,三裂缝,孢子萌发为书带蕨型,原叶体发育为水蕨型,无毛状体产生;培养发现,其配子体能产生精子器,但不产生颈卵器,当接种密度适中时,可进行无配子生殖。（2）在光照和黑暗条件下狭眼凤尾蕨成精子囊素有促进和抑制水蕨孢子萌发的作用,但效果均不显著。（3）在光照条件下,狭眼凤尾蕨成精子囊素可以延迟水蕨心脏形配子体分生组织缺刻的形成,但对其配子体形态和性别分化无明显影响;而在黑暗条件下狭眼凤尾蕨成精囊素对水蕨长条形配子体的形态发育具有一定影响,与对照组相比其顶端分生组织发达,整体呈长楔形,对性别分化影响不显著。可见,狭眼凤尾蕨和水蕨不具有同种成精子囊素系统。     

变异鳞毛蕨的孢子培养与配子体发育研究

应用无菌培养和常规泥土培养两种方法对变异鳞毛蕨(Dryopteris varia)孢子进行了比较研究,并在光学显微镜下观察了其配子体的发育过程.结果表明:蔗糖浓度为2%的1/2MS与MS培养基对孢子萌发时间和萌发率影响不大,但前者较适于孢子萌发,而后者则适于孢子体形成;在1/2MS培养基上,1%的蔗糖浓度比其它浓度更适宜于孢子的萌发.以菜园士为培养基质时,变异鳞毛蕨孢子的萌发时间短且萌发率高,但幼孢子体出现的时间明显晚于无菌培养.孢子萌发为书带蕨型,原叶体发育为三叉蕨型,符合鳞毛蕨属配子体发育的特征.     

重金属铅对3种蕨类植物孢子萌发和配子体发育的影响

研究了重金属铅对水蕨、中华桫椤和扇蕨孢子萌发及其对水蕨配子体发育的影响。结果表明：随着铅浓度的增加,水蕨、中华桫椤和扇蕨孢子的萌发率逐渐降低;在低于10^-4 mol·L^-1的环境中,水蕨孢子可以正常萌发;在低于10^-5 mol·L^-1的环境中,中华桫椤和扇蕨孢子能正常萌发;水蕨、中华桫椤和扇蕨的孢子都对铅具有一定的耐受性;铅对水蕨配子体的发育存在一定影响,铅使水蕨配子体呈不规则的心脏形,多发育为雄配子体;精子器凹陷入原叶体边缘;配子体细胞中叶绿体分布不均匀。本研究为铅污染土壤的植物修复筛选后备植物。     

光照强度对水蕨孢子萌发及配子体性别分化的影响

研究了不同光照强度条件对水蕨孢子萌发以及配子体性别分化的影响。结果表明:2 000~4 000 lx的光照强度条件有利于水蕨孢子萌发,4 000 lx光照条件下孢子萌发最快。光照强度约为4 000 lx时最有利于使水蕨发育为两性配子体。光照强度约为1 000 lx时最有利于使水蕨发育为雄性配子体。为水蕨的人工栽培和分子生物学研究提供理论依据。     

液体培养基条件下乌毛蕨配子体发育的研究

在液体培养基条件下对中国产乌毛蕨(Blechnum orientaleL.)配子体发育进行了观察。结果表明:乌毛蕨的孢子为两面体型,两侧对称,极面观为椭圆形,赤道面观为半圆形或豆形,周壁具脊状褶皱;孢子萌发为书带蕨型;原叶体发育为三叉蕨型,成熟原叶体为对称的心脏形。经比较分析,蕨类植物孢子繁殖时采用混合土培养较适宜;液体培养基和混合土在研究蕨类植物配子体发育时同样具有可行性。孢子形态和配子体发育的观察结果表明,乌毛蕨是蕨类植物中较进化的种类;乌毛蕨科与鳞毛蕨科亲缘关系较近。     

孢子大小和培养基对水蕨孢子萌发及配子体性别分化的影响

利用MS培养基、改良Knop’s培养基、自来水和蒸馏水分别培养水蕨中等大小孢子,同时利用改良Knop’s培养基培养不同大小的水蕨孢子,观察记录不同条件下水蕨孢子萌发和性别分化情况。实验表明,二级孢子（赤道轴120～140μm）萌发率最高;一级孢子（赤道轴〉140μm）萌发最有利于使水蕨发育为两性配子体,三级孢子（赤道轴〈120μm）萌发最有利于使水蕨发育为雄配子体;MS培养基和改良Knop’s培养基相对于自来水和蒸馏水有利于水蕨孢子萌发;各培养基中水蕨两性配子体比率排序是MS培养基〉改良Knop’s培养基〉自来水〉蒸馏水,而雄配子体比率排序与之相反。此结果为水蕨的引种保护、人工繁育和分子生物学研究提供理论依据。     

除草剂二氯喹啉酸对粗梗水蕨有性繁殖的影响

研究了除草剂二氯喹啉酸对国家二级重点保护野生植物粗梗水蕨(Ceratopteris pteridoides)有性繁殖的影响。研究发现二氯喹啉酸对粗梗水蕨孢子萌发没有影响,但对粗梗水蕨配子体生长有抑制作用,二氯喹啉酸浓度和粗梗水蕨配子体面积呈明显剂量-效应关系,二氯喹啉酸对粗梗水蕨配子体生长抑制的EC50为0.48 mg.L-1。随着二氯喹啉酸浓度从0.1 mg.L-1开始升高,粗梗水蕨配子体分化形成的雌雄同体配子体逐渐减少,同时雌雄同体配子体上颈卵器形成时间推迟,颈卵器数量减少。当二氯喹啉酸浓度为4 mg.L-1时,没有雌雄同体配子体形成。以上研究结果表明,粗梗水蕨配子体生长受到二氯喹啉酸的抑制,自身的成精子囊素性别调控系统间接受到影响,导致粗梗水蕨有性繁殖能力减弱。该研究可为除草剂对非靶标水生植物的生态风险评价提供基础资料,保护水生植物多样性。     

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。     

阔叶鳞盖蕨和粗毛鳞盖蕨(碗蕨科)配子体发育的研究

以腐殖土为基质,对阔叶鳞盖蕨Microlepia patyphylla（D．Don）J．Sm．和粗毛鳞盖蕨M.strigosa（Thunb．）Presl进行了孢子繁殖;利用光学显微镜观察和记录了它们的孢子萌发和配子体发育过程。结果表明,两者的孢子及配子体性状极为相似：孢子同型,三裂缝,萌发慢;孢子萌发需要光,萌发方式为书带蕨型Vittaria-type;配子体生长慢,发育类型为铁线蕨型Adiantum—type;原叶体心形或其他形状,无毛状体,多数为雌雄异株。性器类型为薄囊蕨型Leptosporangiate-type。它们既具有大量的原始性状,也具有少数进化性状。与阔叶鳞盖蕨相比,粗毛鳞盖蕨的配子体发育特征更为原始。说明粗己鳞盖蕨的系统位置更加低下。首次观察到阔叶鳞盖蕨中细胞自然死亡时的叶绿体聚集现象。粗毛鳞盖蕨原叶体老化时叶绿体呈现规则的、相互镶嵌的多边形的形状以及在正常光照下粗毛鳞盖蕨某些细胞中的部分叶绿体成链珠状排列等现象。     

三角鳞毛蕨配子体发育的研究

采用混合土培养三角鳞毛蕨(Dryopteris subtriangularis)孢子,显微镜下观察记录其孢子萌发及配子体发育过程。结果表明：孢子深褐色,赤道面豆形,极面观椭圆形,单裂缝。播种1周左右孢子萌发,萌发类型为书带蕨型,配子体发育为三叉蕨型。播种2周左右发育为片状体。播种4周左右发育形成幼原叶体,成熟原叶体呈心脏形。原叶体边缘及腹面均可产生毛状体,数量丰富,为单细胞。播种6周左右开始有性器官出现,精子器近圆球形,由3细胞组成,成熟颈卵器颈部由4列细胞组成,3～5层细胞高。原叶体受精后1个月内即可观察到从原叶体上生成的幼胚。     

麦秆蹄盖蕨（Athyrium fallaciosum）配子体的发育规律及播种密度对配子体性别分化的影响

在组织培养条件下,对麦秆蹄盖蕨（Athyrium fallaciosum）配子体发育的连续过程进行了详细观察。结果表明:麦秆蹄盖蕨孢子为四面体型; 孢子萌发为书带蕨型（Vittaria-type）;原叶体发育为铁线蕨型（Adiantum-type）,成熟原叶体为对称的心形;精子器近圆球形,成熟颈卵器细长,常向原叶体基部倾斜或弯曲。常规播种条件下,发现麦秆蹄盖蕨配子体有雌配子体、雄配子体、雌雄同体配子体和无性配子体类型。配子体的性别随密度不同而呈现一定的变化趋势,雄配子体随密度增大呈上升趋势;雌配子体随密度增大先上升后下降;雌雄同体配子体和无性配子体随密度变化不大。雌配子体和雌雄同体配子体具颈卵器数目一般为10～15个;精子器数目随密度的增大逐渐减少,雄配子体中具有约50个精子器,雌雄同体配子体具有约20个精子器。     

Factors Influencing Spore Germination and Early Gametophyte Development in Anemia mexicana and Anemia phyllitidis

Spores of Anemia mexicana Klotzsch and Anemia phyllitidis (L.) Swartz were tested comparatively to investigate the effects of various treatments on spore germination and early gametophyte development in light and darkness. The optimum pH for induction of spore germination is approximately 6. Both species have a minimum 8 hour light insensitive preinduction phase for spore germination. An additional 8 to 12 hours of light are needed to induce 50% germination in A. phyllitidis while at least 24 hours of light are needed for A. mexicana spores. A. phyllitidis has greater sensitivity to the four gibberellic acids tested (GA₃, GA₄, GA₇, and GA₁₃) than A. mexicana for induction of spore germination in darkness. In both species the greatest response was observed with GA₄ and GA₇. GA₁₃ was clearly the least effective. Gametophytes of each species are 100 times more sensitive to their own antheridiogen than to the antheridiogen of the other species. AMO-1618 (1 millimolar), fenarimol (1 mm), and ancymidol (0.1 mm) had essentially no effect on light-induced germination. The latter two did, however, inhibit gametophyte development.     

短柄禾叶蕨配子体发育的研究

首次通过人工培养的方式,详细记录了短柄禾叶蕨（Grammitis dorsipila (Christ.)C.Chr.）具代表性的配子体形态发育全过程及各环节的特征,包括孢子萌发、原丝体、毛状体、叶绿体、边缘细胞、精子器等,并附特征性结构照片29幅。为禾叶蕨科（Grammitidaceae）的研究积累了配子体发育方面的详实资料,并初步讨论了短柄禾叶蕨配子体特征所体现的系统学意义。     

Antheridiogen,dark spore germination,and outcrossing mechanisms in Bommeria (Adiantaceae)

Isozymic analyses of the patterns of genetic variability in sporophyte populations have demonstrated that most fern species have outcrossing breeding systems. However, because fertilization takes place during the ephemeral, diminutive gametophyte generation, direct observation of breeding systems in nature has not been possible. Recent discoveries of soil-bound spore banks suggested that genetic diversity could be stored beneath the surface and subsequently released by appropriate chemical cues. Previous studies demonstrated that Bommeria sporophytes are the product of outcrossing, that their gametophytes carry high levels of genetic load, and that they produce and respond to antheridiogen. Research reported here demonstrated that Bommeria spores can survive long-term storage but will not germinate in the dark. Antheridiogen, however, will release spores from this light requirement and stimulate germination. Higher concentrations of antheridiogen result in higher germination rates. Gametophytes grown in the dark on antheridiogen-enriched agar form antheridia and release actively swimming sperms. Thus, spores housed beneath the soil surface could remain dormant until stimulated to germinate by antheridiogen secreted by surface-dwelling, archegoniate gametophytes. Sperm released from these subterranean gametophytes could fertilize eggs on the surface. Because spores housed in the soil are likely to be genetically different than those at the surface, heterozygous sporophytes would be more likely to result. Discovering that Bommeria species contain all of the prerequisites for this proposed outcrossing mechanism provides an explanation for the maintenance of genetic diversity in some fern populations.     

沙枣和柠条种子萌发期耐盐性研究

采用室内控制实验的方法,系统研究了沙枣和柠条种子萌发、生长、活力以及生理过程;对萌发、生长及活力指标与盐胁迫进行相关与回归分析,发现了实验室条件下沙枣和柠条种子萌发期的耐盐临界值;对生理指标与盐胁迫,生理指标与相应的萌发及生长指标进行相关性分析,在生理水平对沙枣和柠条种子萌发期对盐胁迫的响应进行了解释。主要结果包括：(1)胚根相对干重含水量与盐胁迫无相关性,初始萌发时间、休眠率以及死亡率与盐胁迫呈显著正相关,其它指标均与盐胁迫呈显著负相关;(2)胚根相对干重含水量与萌发种子初期生长无相关性,休眠率、死亡率与萌发率呈显著负相关,其它指标均与初期生长呈显著正相关;(3)受盐胁迫的抑制程度：萌发速度>萌发启动>萌发率;胚根生长>胚轴生长>萌发率;(4)种子萌发期,沙枣耐盐性强于柠条,两者耐盐临界值均高于0.1 mol·L^-1。     

Specificity of the Antheridogen from Ceratopteris thalictroides (L.) Brongn

Characteristics of the fern antheridogen from Ceratopteris thalictroides (L.) Brongn. are investigated. These are: (a) determination of molecular size (it is readily dialyzable), (b) pK_a (about 5), (c) movement in thin layer chromatography, and (d) ability to induce dark germination of fern spores. These four characteristics are compared to the same characteristics of three other antheridogens (antheridogens A and B or GA). Molecular size and pK_a are similar, but, the antheridogens are separable from each other using thin layer chromatography. It was also shown that spore germination is not induced by the Ceratopteris antheridogen, even in its own spores, a characteristic not reported as shared by the other antheridogens. However, the inconsistency of spore germination as an assay for antheridogen is demonstrated. The presence of gametophyte-produced allelopathic substances is also shown.     

FERN GAMETOPHYTE DEVELOPMENT: CONTROLS OF DIMORPHISM IN CERATOPTERIS THALICTROIDES

Multispore cultures of Ceratopteris thalictroides (L.) Brongn. gametophytes contain plants of two distinct morphologies, hermaphrodite and male (Fig. 3). The male gametophyte has previously been shown to be induced by an antheridogen system. In the absence of the antheridogen all gametophytes become hermaphroditic. It is shown that the hermaphroditic gametophytes are first to develop in multispore cultures and are also the only source of antheridogen. Thus, since males are only produced in the presence of antheridogen, an explanation of the occurrence of two morphologies is evident. That is, the first gametophytes to develop do so in the absence of antheridogen, become hermaphrodites and produce antheridogen to which later developing gametophytes respond by becoming males. This does not explain why there is a range in development times for the gametophytes. Three possible controlling factors of development are investigated in multispore cultures: spore size, time of germination, and gametophyte growth. Data collected on these three factors are statistically analyzed. Analyses were done to determine the relative importance (singly or in combination) of these factors in predicting the developmental potential of a gametophyte. It was found that spore size most accurately indicated (ca 75% of the time) future gametophyte development.     

Spore Germination of Ceratopteris thalictroides (L.) Brongn

Germination of the spores of Ceratopteris thalictroides wasexamined by light and scanning electron microscopy. Spores germinateby scission of the spore coat at the trilete markings. The initialcell divisions produce a proximal prothallial initial with rhizoidsthat are distal and lateral. This represents a reverse orientationof the more common situation in ferns. Evidence is presentedthat rhizoid initials divide with the most lateral of the daughtercells differentiating into a rhizoid. Rhizoids are long andunbranched. The uniseriate protonema is transient with bi-dimensionalgrowth established quite early. About a week following germination,the gametophyte is broad at the basal region and narrow at theanterior region. Further growth of cells in the basal regionis by elongation and the anterior region broadens to producean elongate-ovate gametophyte.     

日本蹄盖蕨配子体发育的研究

采用混和土培养日本蹄盖蕨(Athyrium niponicum)孢子,显微镜下观察记录其孢子萌发及配子体发育过程。结果表明:孢子黑褐色,赤道面豆形,极面观椭圆形,单裂缝。播种7 d左右孢子萌发,萌发类型为向心型,配子体发育为铁线蕨型。丝状体7~11细胞时开始发育为片状体。播种14 d后发育形成幼原叶体,成熟原叶体呈心脏形。原叶体边缘可产生单细胞毛状体。播种后20 d左右精子器出现,精子器近圆球形,由3细胞组成。7 d后颈卵器出现,成熟颈卵器3~5层细胞高。精卵受精后14 d左右即可观察到从原叶体上生成的幼胚。