PHOTOMORPHOGENESIS OF THE GAMETOPHYTES OF LYGODIUM JAPONICUM

Protonemata of Lygodium japonicum turn biplanar in both red and blue light regimes and remain filamentous in far-red light. Biplanar gametophytes formed in red light are longer than broad with long, rectangular cells, whereas in blue light they appear broader than long with short, isodiametric cells. Transfer of protonemata of all ages from far-red regime to red or blue light induces a morphological type of growth characteristic of the new light regime. However, only relatively young biplanar forms transferred from red or blue light are able to resume filamentous type of growth in a subsequent regime of far-red light.     

Comparative development of heavily asymmetric-cordate gametophytes of <Emphasis Type="Italic">Anemia phyllitidis</Emphasis> (Anemiaceae) focusing on meristem behavior

Development of heavily asymmetric cordate gametophytes of Anemia phyllitidis (Anemiaceae), one of the schizaeoid ferns, was examined using a sequential observation technique; epi-illuminated light micrographs of the same growing gametophytes were taken approximately every 24 h. The apical cell-like wedge-shaped cell was produced once from the terminal cell of a germ filament, but it stopped dividing soon after production of one or two derivative cells. Without a functional apical cell, the gametophyte developed by intercalary growth until the early stage of wing formation, and then the multicellular (pluricellular) meristem arose from the lower lateral side of the gametophyte. This was in sharp contrast to the observation that the multicellular meristem forms in place of the apical cell in typical cordate gametophytes. Loss of the functional apical cell probably caused a site-shift in the multicellular meristem of the Anemia phyllitidis gametophyte during evolution from apical to lateral. The results suggest that apical cell-based and multicellular meristems are primarily independent of each other. The multicellular meristem produced cells equally in the distal and proximal directions to form wings in both directions but proximally produced cells divided much less frequently. As a result, a heavily asymmetric gametophyte was formed.     

<Emphasis Type="Italic">In vitro</Emphasis> culture of <Emphasis Type="Italic">Drynaria fortunei</Emphasis>, a fern species source of Chinese medicine “Gu-Sui-Bu”

This study reports spore germination, early gametophyte development and change in the reproductive phase of Drynaria fortunei, a medicinal fern, in response to changes in pH and light spectra. Germination of D. fortunei spores occurred on a wide range of pH from 3.7 to 9.7. The highest germination (63.3%) occurred on ½ strength Murashige and Skoog basal medium supplemented with 2% sucrose at pH 7.7 under white light condition. Among the different light spectra tested, red, far-red, blue, and white light resulted in 71.3, 42.3, 52.7, and 71.0% spore germination, respectively. There were no morphological differences among gametophytes grown under white and blue light. Elongated or filamentous but multiseriate gametophytes developed under red light, whereas under far-red light gametophytes grew as uniseriate filaments consisting of mostly elongated cells. Different light spectra influenced development of antheridia and archegonia in the gametophytes. Gametophytes gave rise to new gametophytes and developed antheridia and archegonia after they were transferred to culture flasks. After these gametophytes were transferred to plastic tray cells with potting mix of tree fern trunk fiber mix (TFTF mix) and peatmoss the highest number of sporophytes was found. Sporophytes grown in pots developed rhizomes.     

Developmental morphology of strap-shaped gametophytes of Colysis decurrens: a new look at meristem development and function in fern gametophytes

Background and Aims

The gametophytes of most homosporous ferns are cordate–thalloid in shape. Some are strap- or ribbon-shaped and have been assumed to have evolved from terrestrial cordate shapes as an adaptation to epiphytic habitats. The aim of the present study was to clarify the morphological evolution of the strap-shaped gametophyte of microsoroids (Polypodiaceae) by precise analysis of their development.

Methods

Spores of Colysis decurrens collected in Kagoshima, Japan, were cultured and observed microscopically. Epi-illuminated micrographs of growing gametophytes were captured every 24 h, allowing analysis of the cell lineage of meristems. Light microscopy of resin-sections and scanning electron microscopy were also used.

Key Results

Contrary to previous assumptions that strap-shaped Colysis gametophytes have no organized meristem, three different types of meristems are formed during development: (1) apical-cell based – responsible for early growth; (2) marginal – further growth, including gametophyte branching; and (3) multicellular – formation of cushions with archegonia. The cushion is two or three layers thick and intermittent. The apical-cell and multicellular meristems are similar to those of cordate gametophytes of other ferns, but the marginal meristem is unique to the strap-shaped gametophyte of this fern.

Conclusions

The strap-shaped gametophytes of C. decurrens may have evolved from ancestors with a cordate shape by insertion of the marginal meristem phase between the first apical-cell-based meristem and subsequent multicellular meristem phases. Repeated retrieval of the marginal meristem at the multicellular meristem phase would result in indefinite prolongation of gametophyte growth, an ecological adaptation to epiphytic habitats.     

铁角蕨属4种植物配子体发育的比较研究

以腐叶土为基质培养铁角蕨(Asplenium trichomanes L.)、阿尔泰铁角蕨[A.altajense(Kom.)Grubov]、假大羽铁角蕨(A.pseudolaserpitii folium Ching)和细裂铁角蕨(A.tenui folium D.Don)的孢子,用光学显微镜观察并比较它们的配子体发育过程,以期为铁角蕨属的系统学研究提供基础资料.结果显示,4种铁角蕨属植物配子体均具有:孢子两面型、单裂缝、周壁具褶皱、书带蕨型萌发,成熟原叶体心形,原叶体具有毛状体等共同特征,表明铁角蕨属的演化处于较进化的系统位置,但它们的毛状体形态和细胞数目明显不同,可为属内分系提供依据.研究发现,4种铁角蕨属植物配子体的边缘细胞形状以及假根的形态和数量均有差异,其叶绿体能象单细胞一样进行无丝分裂,强光照射下叶绿体向相邻细胞的侧壁集中等现象.     

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。     

Attractive and repulsive interactions between female and male gametophytes in Arabidopsis pollen tube guidance

Sexual reproduction in plants, unlike that of animals, requires the action of multicellular haploid gametophytes. The male gametophyte (pollen tube) is guided to a female gametophyte through diploid sporophytic cells in the pistil. While interactions between the pollen tube and diploid cells have been described, little is known about the intercellular recognition systems between the pollen tube and the female gametophyte. In particular, the mechanisms that enable only one pollen tube to interact with each female gametophyte, thereby preventing polysperm, are not understood. We isolated female gametophyte mutants named magatama (maa) from Arabidopsis thaliana by screening for siliques containing half the normal number of mature seeds. In maa1 and maa3 mutants, in which the development of the female gametophyte was delayed, pollen tube guidance was affected. Pollen tubes were directed to mutant female gametophytes, but they lost their way just before entering the micropyle and elongated in random directions. Moreover, the mutant female gametophytes attracted two pollen tubes at a high frequency. To explain the interaction between gametophytes, we propose a monogamy model in which a female gametophyte emits two attractants and prevents polyspermy. This prevention process by the female gametophyte could increase a plant's inclusive fitness by facilitating the fertilization of sibling female gametophytes. In addition, repulsion between pollen tubes might help prevent polyspermy. The reproductive isolations observed in interspecific crosses in Brassicaceae are also consistent with the monogamy model.     

AN ULTRASTRUCTURAL STUDY OF FERN GAMETOPHYTES DURING ONE- TO TWO-DIMENSIONAL DEVELOPMENT

During early stages in the transition from 1- to 2-dimensional gametophyte development the change from filamentous to bulbous apical cell is not accompanied by major changes in the nature or number of cytoplasmic components of the cell. However, chloroplasts in apical cells of plants grown in red light are larger than those from cells of plants grown in blue light. In addition, the orientation of cytoplasmic microtubules is different in apical cells of plants from red and blue light. This change in orientation may be causally related to the change in apical cell form during 1- to 2-dimensional growth.     

Negative phototropism in young gametophytes of the fern Schizaea pusilla

Schizaea pusilla is a rare fern that occurs in acidic bogs and is one of the few fern species that maintains a filamentous gametophyte throughout its development. To expand our knowledge of the physiology of this fern, phototropic responses were examined in young gametophytes. In contrast to germ filaments of other fern species, apical protonemata of young gametophytes are negatively phototropic in continuous white, red and blue light at all fluence rates tested. The expression of phototropic curvature is not limited by time since apical protonemata are also negatively phototropic when they are given brief exposures of light and then placed in the dark. In other lower plant groups such as mosses and some algae, the direction of phototropic curvature can change depending on light quality and intensity, but in young gametophytes of Schizaea, negative phototropic curvature was observed in all conditions studied. Blue light is the most effective in promoting the negative phototropic response in Schizaea.     

Gametophyte development and reproduction of <Emphasis Type="Italic">Argyrochosma nivea</Emphasis> (Pteridaceae)

The gametophyte of Argyroschosma nivea was studied, mainly focusing in its morphological development, and in the apogamous production of sporophytes. Some observations on the spores were also made. As far as it is known, this is the second species of the genus whose gametophytes are studied. The germination pattern followed the Vittaria type. The subsequent developmental processes followed the Ceratopteris type. Some of the gametophytes reached an adult stage with a cordate, symmetric shape, but most of them developed as irregular, lobed prothalli. The sporophyte emerged from the anterior part of the prothallus, without formation of gametangia. First, a cell became active and originated a proliferating area of small cells. From this area, long glandular hairs were formed followed by a projected conical cluster of cells. The cluster elongated into a sporophytic structure and its apex became progressively spatulate and finally trilobulate, with marginal, glandular hairs, stomata and tracheids continuously produced. This sporophyte secreted granules of white farina from its beginnings. The production of farina in the sporophyte but not in the gametophyte could help to support the idea of the segregation of this species from its traditional location in Notholaena to Argyrochosma, as farinose gametophytes seem to be a synapomorphy of the notholenoids, group that includes Notholaena but not Argyroschoma.     

银粉背蕨配子体及幼孢子体发育的研究

银粉背蕨是一种小型观赏蕨类植物,但目前我国对该蕨的研究还不够成熟。本文利用改良Knop's培养基和腐殖土培养银粉背蕨的孢子,观察其配子体及幼孢子体形态发育特征,并研究了其配子体发育的最适培养基pH值。研究结果显示：（1）银粉背蕨孢子黄褐色,具三裂缝,极面观三角圆形,赤道面观为近半圆形,孢子具网状纹饰;孢子萌发为书带蕨型;原叶体发育为水蕨型;颈卵器和精子器为薄囊蕨型;成熟原叶体为对称的心脏形,不具毛状体;上述特征为银粉背蕨孢子和配子体发育的稳定特征。（2）培养基pH值在7.0~9.0时随着碱性的增强,银粉背蕨孢子萌发和配子体生长发育速度逐渐增加。（3）利用腐殖土培养银粉背蕨孢子,7~8周可发育成幼叶,成苗率达90%,成苗健壮,根系发达,是扩繁银粉背蕨的适宜方式。本文为资源植物银粉背蕨人工繁殖和演化研究提供科学依据。     

Blue light effects on rose photosynthesis and photomorphogenesis

Through its impact on photosynthesis and morphogenesis, light is the environmental factor that most affects plant architecture. Using light rather than chemicals to manage plant architecture could reduce the impact on the environment. However, the understanding of how light modulates plant architecture is still poor and further research is needed. To address this question, we examined the development of two rose cultivars, Rosa hybrida‘Radrazz’ and Rosa chinensis‘Old Blush’, cultivated under two light qualities. Plants were grown from one‐node cuttings for 6 weeks under white or blue light at equal photosynthetic efficiencies. While plant development was totally inhibited in darkness, blue light could sustain full development from bud burst until flowering. Blue light reduced the net CO₂ assimilation rate of fully expanded leaves in both cultivars, despite increasing stomatal conductance and intercellular CO₂ concentrations. In ‘Radrazz’, the reduction in CO₂ assimilation under blue light was related to a decrease in photosynthetic pigment content, while in both cultivars, the chl a/b ratio increased. Surprisingly, blue light could induce the same organogenetic activity of the shoot apical meristem, growth of the metamers and flower development as white light. The normal development of rose plants under blue light reveals the strong adaptive properties of rose plants to their light environment. It also indicates that photomorphogenetic processes can all be triggered by blue wavelengths and that despite a lower assimilation rate, blue light can provide sufficient energy via photosynthesis to sustain normal growth and development in roses.     

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。     

Life-cycle of <Emphasis Type="Italic">Scinaia interrupta</Emphasis> (Nemaliales,Rhodophyta)

The life-cycle of Scinaia interrupta (A.P. de Candolle) M. J. Wynne was investigated in vitro using four irradiance regimes: 4, 8, 12 and 16 μmol photons m⁻² s⁻¹. A triphasic heteromorphic life-cycle was observed. Carpospores released by cystocarps of gametophytes collected in the field developed into filamentous tetrasporophytes, which produced tetrahedral tetrasporangia. Tetrasporangial development was accelerated under higher irradiance levels. Tetraspores germinated into filamentous protonemal gametophytes, initially identical to the tetrasporophyte. Filamentous gametophytes developed apical utricles and gave rise directly to the fleshy gametophyte. Further development of the fleshy gametophyte was not observed at the lowest irradiance regime (4 μmol photons m⁻² s⁻¹). The present study reports for the first time the influence of the irradiance regime on the initial tetrasporangial development and in the development of the fleshy gametophyte, and reinforces the importance of light intensity on Scinaia life-cycle. Production of apical utricles by the filamentous gametophyte is newly reported for the genus.     

INTERPOPULATIONAL VARIATION IN THE GAMETOPHYTES OF PELLAEA ANDROMEDAEFOLIA

The development and mature morphology of the gametophytes from both sexual and apogamous populations of the fern Pellaea andromedaefolia were investigated. While most sexual examples were indistinguishable, some differences were noted. An insular collection was distinctive in its variability and irregularity of form. Although the latter was a representative of var. pubescens, other collections of the variety could not be distinguished from var. andromedaefolia on the basis of gametophytic characteristics. The apogamous gametophytes were decidedly more variable in development and often very different from sexual thalli. The mature asexual thalli tended to be more irregular in form and usually sharply divergent from the typical cordate type characteristic of the sexual populations. Each of the five apogamous samples was unique with respect to gametophyte development. The differences among the gametophytes of the various populations do not correlate with the sporophytic characteristics which differentiate the two varieties of the species.     

Growth Promotion of Vegetative Gametophytes of <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> by Blue Light

Through an acclimation period of 10 days, compared to white light, the maximal net photosynthetic rates were significantly higher for gametophytes of Undaria pinnatifida cultivated under blue light (400–500 nm), and were lower under red light (600–700 nm). Chlorophyll c and the carotenoid content of gametophytes were similar under blue light and red light but were much lower under white light. The growth rate of female gametophytes under blue light was higher than that under other lights, and the growth rate of male gametophytes showed little variation with respect to blue and white light. Male and female gametophytes were mixed together to form sporophytes under white, blue and red light. After approximately 5 days, 50% gametophytes became fertile under blue and white light, but remained vegetative under red light after 10 days.     

不同环境因子对珍稀观赏蕨类金毛狗配子体性别分化的影响

金毛狗[Cibotium barometz(L.)J.Sm.]是珍稀观赏蕨类的重要类群,为国家二级重点保护野生植物。该研究以金毛狗孢子为试验材料,探究培养密度、外源赤霉素以及光质等不同环境因子对金毛狗配子体性别分化的影响,为金毛狗人工繁育和蕨类植物配子体性别决定机制研究提供技术支持。结果表明:(1)低配子体培养密度(1个/cm2和5个/cm2)有利于颈卵器和雌配子体形成,随着配子体培养密度增加,颈卵器平均数量及雌配子体比率下降,精子器平均数量以及雄配子体和两性配子体比例增加,但配子体培养密度过高(80个/cm2)会导致大量无性配子体产生。(2)不同配子体培养密度下,随着培养时间延长,两性配子体比率均有增加,且增加幅度基本一致。(3)外源GA4显著抑制颈卵器和雌配子体形成,并显著促进精子器和雄配子体形成;外源GA3对金毛狗配子体性别分化没有显著影响。(4)白光、红光、蓝光等不同光质对金毛狗配子体性别分化未产生显著影响,但会影响配子体的发育和形态建成。     

Antheridogen activity in the fern Ceratopteris thalictroides (L.) Brongn.

Spores of the homosporous fern Ceratopteris thalictroides , in multispore culture, initially produce spatulate gametophytes bearing only antheridia (males) and cordate gametophytes bearing both antheridia and archegonia (hermaphrodites). When multispore cultures are sampled, the ratio of male to hermaphroditic gametophytes is a constant for each population examined. Four possible causes of such a sex ratio (cytoplasmic inheritance, nuclear inheritance, incipient heterospory and an antheridogen) are investigated. Evidence presented indicates that an antheridogen causes the existence of two gametophyte types, while one or more cytoplasmic units are the probable cause of the sex ratio. The activity of the antheridogen is to cause potentially hermaphroditic plants to become male. This activity was elucidated in monospore culture. Populational differences in antheridogen activity are also demonstrated. The significance of antheridogens is discussed in relation to the mating system of these plants.     

Electrical Changes in the Apical Cell of the Fern Gametophyte during Irradiation with Photomorphogenetically Active Light

Electrophysiological procedures were used to evaluate cellular responses of fern (Onoclea sensibilis L.) gametophytes to photomorphogenetically active light. Red, far red, and blue light caused rapid changes in the membrane potential of the apical cell of the gametophyte filament; other cells in the filament were not similarly responsive. Measurements made with one electrode in the apical cell revealed that the membrane potential depolarized in red light and repolarized in far red light. Irradiation with blue light caused a hyperpolarization, the rapidity of which was dependent on a red light pretreatment. More refined measurements with one electrode in the tip of the apical cell and another in the base of the cell showed that both red and blue light treatments cause the apical cell to behave as a dipole. Because of the profound, long-term morphological changes that follow light irradiation in this organism, it was hoped to use it to elucidate the role that electrical parameters play in determining subsequent developmental events.     

Jasmonic acid promotes division of fern protoplasts, elongation of rhizoids and early development of gametophytes

The effects of jasmonic acid (JA) on spore germination, early gametophyte development and sporophytic protoplast culture of the fern Platycerium bifurcalum (Cav.) C. Chr. were investigated. JA and no influence on spore germination and primary rhizoid initiation, but significantly promoted early gametophyte development, which was evident from the longer primary rhizoids as well as the higher number of rhizoids and cells per gametophyte. Jasmonic acid (1 μ M ) also promoted the transition of gametophytes from a filamentous to a planar growth. Optimal primary rhizoid elongation and highest cell division activities in the gametophytes were observed at 0.01–1 μ M JA, while the highest number of rhizoids on gametophytes was obtained at 0.1–1 μ M JA. Jasmonic acid (0.01 μ M ) also stimulated initial protoplast divisions. Except for the experiment in which the effect of JA on germination was tested. JA concentrations exceeding 1 μ M did not promote cell elongation or cell division but were instead inhibitory. On the basis of these findings, we propose that JA may be involved in early stages of P. bifurcatum development.