Ultrastructural analysis of sporoderm development in megaspores ofSelaginella galeottii (Lycophyta)

Structural members within the exospore ofSelaginella galeottii suggestive of those present at maturity are first detectable when the exospore is approximately 5 µm in thickness. Subsequent changes in successively larger sporangia involve a gradual size increase of the component units simultaneously throughout the exospore. Concomitantly, non-membrane bound material present at the inner surface of the tapetum (and the persistent megasporocytes) and throughout the sporangium locule changes from primarily droplets and weftlike material (including beaded wefts) to coarse fibrous material. The taxa which possess this unusual wall pattern cut across presently accepted taxonomic schemes. This is not the case with the other wall ultrastructural types in the genus. The possibility exists that this megaspore wall type defines a separate lineage within the genus which, by virtue of its large megaspores, was able to compete well and radiate to produce a variety of life forms.     

Megaspore wall growth inSelaginella (Lycopodiatae)

Different stages of megaspore and megasporangial development inSelaginella argentea (Wallich)Spring,S. bigelowii Unerw., andS. kraussiana (Kze.)A. Br. have been seen and studied. Megaspore wall units give positive reactions for polysaccharides and protein in young megaspores, and become the thick and resistant wall typical of the genus only later.—Units forming the exospore and the spaces between units enlarge from widths of 5–10nm early during development up to over 200 nm at pregermination stages. The spaces enlarge first. Initially they are circular and mostly about 70 nm in diameter. Later, spaces toward the inner part of the exospore enlarge more than those near the outer surface. During pregermination, wall spaces range in size from 4 to 50 times the width of units with the larger spaces located near the inner surface. As a result the exospore would be under tension to spring outward during germination when the laesurae are lysed.—A gap in the exospore, shaped like a half-moon in polar sections, forms in equatorial and distal portions of the spore. This gap becomes enormous, three times the volume of the central space plus the mesospore, and is filled with lipids and other nutrients. Late in development, during the period of tapetal cell degeneration, the gap contents are moved into the central space and the gap is closed.—Late in development the mesospore is degraded. Its products, along with gap contents, seem to be added to the contents of the central cavity and appear as reserve storage globules. A primary wall-like endospore is formed during this period, at the inner surface of the exospore. During germination this endospore develops further at its inner surface.—Changes in the size and shape of megasporangia occur independently of the size of megaspores.Megaspore development inSelaginella. II. For first part seeMorbelli & Rowley (1993).     

Application of confocal microscopy to exospore ultrastructure in megaspores ofSelaginella convoluta andS. marginata

Information pertaining to the arrangement of the rodshaped units that form the exospore ofSelaginella convoluta (Walk. Arn.) Spring andS. marginata (Humb. & Bonpl.) Spring megaspores was obtalned using both a confocal laser scanning microscope and a transmission electron microscope. Units are helically coiled, as we interpret them. The highest levels of fluorescence with confocal microscopy were in the places where the coiled exospore units that contain the fluorochrome dye overlap. These sites of overlap occurred in a close packed (hexagonal or pentagonal) arrangement. A more-or-less circular central nonfluorescent area was contiguous between overlapping exospore units. We conclude that the space between exospore units is a continuous channel (conduit). Each conduit is embraced by five or six helical-units that interdigitate.     

Isolation and characterization of microsatellite loci of Lycopodium fordii Bak. (Lycopodiaceae, Pteridophyta)

Lycopodium fordii Bak. (Lycopodiaceae, Pteridophyta) is a gardening plant with a native, fragmentary distribution in Taiwan. In this study, we described the development of eleven microsatellite loci in L. fordii for genetic studies. These new markers were tested in 16 individuals of three populations. The number of alleles ranged from 1 to 5 and the expected heterozygosity from 0.41746 to 0.72222. Four of the nine polymorphic loci were significantly deviated from Hardy-Weinberg expectations due to the heterozygote deficiency. The microsatellite markers have also been proved as informative genetic markers for other 15 Lycopodium species.     

Occurrence and evolutionary origins of polyploids in the clubmoss genus Diphasiastrum (Lycopodiaceae)

Two polyploid taxa are commonly recognized in the genus Diphasiastrum, D. wightianum from Asia and D. zanclophyllum from South Africa and Madagascar. Here we present results from Feulgen DNA image densitometry analyses providing the first evidence for the polyploid origin of D. zanclophyllum. Also reported for the first time are data confirming that D. multispicatum and D. veitchii, representing the putative parent lineages of D. wightianum, are diploids. Phylogenetic analyses of the nuclear regions RPB2, LEAFY and LAMB4 reveal that putative tetraploid accessions are of allopolyploid origin. Diphasiastrum zanclophyllum shows close relationship to the North American taxon D. digitatum on the maternal side, but the paternal relationship is less clear. Two accessions from Asia, both inferred to be polyploid, have D. veitchii as the maternal parent, whereas the paternal paralogs show relationships to D. multispicatum and D. tristachyum, respectively. None of these parental combinations have previously been hypothesized.     

中国马尾杉属(石松科)的分类修订

马尾杉属植物因含治疗阿尔茨海默症的特效药成分石杉碱甲而具有重要的经济价值和保护价值，全属为国家重点保护植物。该文基于形态学、生态学和地理学等证据，对中国分布的马尾杉属植物进行了分类修订，承认我国有马尾杉属植物21种，将该属植物划分为4个单系的组，即金丝条马尾杉组(Sect.Fargesiani X. C. Zhang&R. H. Jiang, sect. nov.)、喜马拉雅马尾杉组(Sect.Hamiltoniani C. Y. Yang, emend. X. C. Zhang&R. H. Jiang)、马尾杉组(Sect.Phlegmariurus)和粗糙马尾杉组[Sect.Squarrosurus(Herter) X. C. Zhang&R. H. Jiang, comb.&stat. nov.],并将Huperzia medogensis、Phlegmariurus austrosinicus、P.changii、P.nylamensis、P.cancellatus var.minor、P.qiongzhongensis和P.shangsiens...     

New species of Selaginella (Selaginellaceae) from the Guayana Highland of Venezuela

Three new species of Selaginella are described from the Guayana Highland of Venezuela: S. cardiophylla, S. hemicardia and S. pruskiana. They are illustrated and the relationships of each are discussed. Another species, provisionally determined as S. terezoana, is reported for the first time from Guyana.     

Development of embryo and endosperm inEragrostis unioloides (Poaceae)

The embryo development conforms to the Asterad type or Megarchetype II. A single cotyledon called scutellum is derived from the entire tierl. The shoot apex, lateral in position, and the coleoptile, originate from the tierl. The remaining parts of the mature embryo are derived from the tiers situated belowl. The short suspensor takes its origin from the lowermost tierp. The endospermab initio is free nuclear in development. It becomes cellular first around the proembryo. Physiologically, the ripe endosperm represents a dead tissue.     

The fine structure and development of the trabeculae and the trabecular ring in Selaginella kraussiana

In the shoot apex of Selaginella kraussiana A. Br., air spaces develop between the endodermal cells, isolating the two steles from the cortex. The endodermal cells elongate to form trabeculae connecting the two tissues. As each trabecular cell develops, cuticularization occurs on the external surface at a point mid-way along the cell and forms the trabecular ring. The plasmalemma of the trabecular cell becomes attached to the wall beneath the main concentration of the ring, resembling the connection between the plasmalemma and the Casparian strip of a primary endodermis. The trabeculae retain a functional cytoplasm throughout growth.     

Floral development in Adonideae (Ranunculaceae)

Floral development and floral phyllotaxis in species of Adonis, Callianthemum, and Trollius (Ranunculaceae) were studied with scanning electron microscopy. The floral organs are initiated in spiral sequence and the flowers have spiral phyllotaxis. The sepal primordia are broad, crescent-shaped, and truncate, but those of petals, stamens, and carpels are rather hemispherical. A relatively long plastochron appears to be present between the last sepal and the first petal as compared with the short and equal plastochrones of all subsequent floral organs. Maturation of the stamens within the androecium appears to be centripetal. The carpels have a short ascidiate zone. Placentation is uniformly lateral, even in Adonis and Callianthemum, which have only one fertile ovule per carpel (versus median in other genera of Ranunculoideae with a single fertile ovule). In Adonis and Callianthemum at the tip of the carpel the ventral slit is gaping and the stigma is broadly exposed, whereas in Trollius the stigma is narrower and more pronouncedly decurrent along the ventral slit. The petals in Callianthemum and Trollius are more conspicuously delayed in development than those in Adonis as compared with sepals and stamens. A short carpel stipe is formed early in Callianthemum but later in Adonis and Trollius. In Trollius farreri (commonly having only five carpels in contrast to other species of Trollius) the carpels form a single (spiral) series. Thus floral development is similar in all three genera and, at a lower level, Adonis and Callianthemum are especially close but have different autapomorphies, which reflects the current classification of the genera.     

Preliminary assessment of somatic cell nuclear transfer in the dromedary (Camelus dromedarius)

Somatic cloning may enable the maintenance/expansion of the population of camels with the highest potential for milk production or the best racing performances. However, there have been no reports of embryonic or somatic nuclear transfer in camels. The aim of this study was to produce dromedary embryos by nuclear transfer using in vitro matured oocytes and two somatic cells from two sources (adult fibroblasts or granulosa cells). A total of 58 adult females were superstimulated by a single dose of eCG (3500 IU). Ten days later, their ovaries were collected postmortem. Cumulus–oocytes-complexes (COCs) were aspirated from stimulated follicles and were matured in vitro for 30 h. Fibroblasts (from live adult male) and granulosa cells (from slaughtered adult females) were used as donor karyoplasts and injected into mature enucleated dromedary oocytes.The cleavage rate was significantly higher (P < 0.05) for embryos reconstructed with fibroblasts (59%) versus those with granulosa cells (45%). However, there was no difference between the two groups in the proportion of cloned embryos reaching the blastocyst stage (fibroblasts: 14% vs. granulosa cells: 15%) or those that hatched (fibroblasts: 10% vs. granulosa cells: 12%). The viability of reconstructed dromedary embryos from the two sources of donor cells (fibroblasts; n = 5 vs. granulosa cells; n = 7) was examined by transferring them to synchronized recipients. Two females (fibroblasts: 1/5; 20%, granulosa cells: 1/7; 14%) were confirmed pregnant by ultrasonography at 15 and 25 days following transfer. Later, the pregnancies were followed by pregnancy empirical-symptoms. These two pregnancies were lost between 25 and 60 days following transfer, respectively.In conclusion, the present study shows for the first time that the development of dromedary NT embryos derived from either adult fibroblasts or granulosa cells can occur in vitro and the transfer of these cloned embryos to recipients can result in pregnancies.     

Comparing infant and juvenile behavior in bonobos (Pan paniscus) and chimpanzees (Pan troglodytes): a preliminary study

The dichotomy between the two Pan species, the bonobo (Pan paniscus) and chimpanzee (Pan troglodytes) has been strongly emphasized until very recently. Given that most studies were primarily based on adult individuals, we shifted the “continuity versus discontinuity” discussion to the infant and juvenile stage. Our aim was to test quantitatively, some conflicting statements made in literature considering species differences between immature bonobos and chimpanzees. On one hand it is suggested that infant bonobos show retardation in motor and social development when compared with chimpanzees. Additionally it is expected that the weaning process is more traumatic to chimpanzee than bonobo infants. But on the other hand the development of behaviors is expected to be very similar in both species. We observed eight mother–infant pairs of each species in several European zoos. Our preliminary research partially confirms that immature chimpanzees seem spatially more independent, spending more time at a larger distance from their mother than immature bonobos. However, the other data do not seem to support the hypothesis that bonobo infants show retardation of motor or social development. The development of solitary play, environmental exploration, social play, non-copulatory mounts and aggressive interactions do not differ between the species. Bonobo infants in general even groom other group members more than chimpanzee infants. We also found that older bonobo infants have more nipple contact than same aged chimpanzees and that the weaning process seems to end later for bonobos than for immature chimpanzee. Additionally, although immature bonobos show in general more signs of distress, our data suggest that the weaning period itself is more traumatic for chimpanzees.     

Ultrastructural study of spore wall morphogenesis inOphioglossum thermale kom. var.nipponicum (Miyabe et Kudo) nishida

Spore wall morphogenesis ofOphioglossum thermale var.nipponicum was examined by transmission electron microscopy. The spore wall of this species consists of three layers: endospore, exospore, and perispore. The spore wall development begins at the tetrad stage. At first, the outer undulating lamellar layer of the exospore (Lo) is formed on the spore plasma membrane in advance of the inner accumulating lamellar layer (Li) of the exospore. Next, the homogeneous layer of the exospore (H) is deposited on the outer lamellar layer. Both lamellar layers may be derived from spore cytoplasm; and the homogeneous layer, from the tapetum. Then the endospore (EN) is formed. It may be derived from spore cytoplasm. The membranous perispore (PE), derived from the tapetum, covers the exospore surface as the final layer. Though the ornamentation of this species differs distinctly from that ofO. vulgatum, the results mentioned above are fundamentally in accordance with the data obtained fromO. vulgatum (Lugardon, 1971). Therefore, the pattern of spore wall morphogenesis appears to be very stable in the genusOphioglossum.     

Structure of wall layers in Selaginella kraussiana microspores (Lycophyta)

A similarity was found in both construction and ultrastructure between the two exospore layers in microspores of Selaginella kraussiana. The exospore is made up of two different kinds of rods. One of the kinds of rods are large, 100–150 nm in width, while the other are tubular rods 10–15 nm in diameter. The large rods are wider at the base of the spines than in the upper part, possibly due to flattening or compression. Both the outer and the inner exospores have a stranded surface that is very pronounced in the microspores of this species. Fibrous strands persisting the scanning electron microscope and transmission electron microscope (TEM) fixations were observed on the spore surface proximally and through perforations (exospore channel openings). This net of fibres penetrates and fills the space of the cavities within large channels through the outer and inner exospore and within the gap. According to our interpretation, these strands would be produced by the tapetum and are probably related to the nourishment of the developing microspores. Contrast varies in TEM sections after cytochemical stains, but this appears to be due to transitory substances, e.g. carbohydrates, rather than to be a substantial difference in basic composition between inner and outer exospore layers.     

海南七仙岭7种卷柏属植物的微观形态与生境间的关系分析

为探讨生境对卷柏属(Selaginella)植物微观形态的影响,利用扫描电子显微镜对海南七仙岭采集的7种卷柏属植物的侧叶、中叶、孢子叶的叶表皮形态以及小孢子形态进行全面观察分析,并计算气孔器大小、气孔密度、孢子大小等,比较分析其微观形态的区别及微形态与生境间的关系,为卷柏属植物的分类提供依据。结果显示:(1)同种卷柏属植物的侧叶、中叶与孢子叶在叶表皮形态上具明显差异,尤其是孢子叶上的气孔与营养叶相对较小且稀疏,与孢子叶的繁殖功能相符合。(2)不同种卷柏属植物的叶表皮特征也明显不同,主要表现在叶缘刺、气孔和瘤状突起上,表明这些特征可以作为卷柏属植物种间区分的依据。(3)卷柏属植物的小孢子形态稳定,纹饰多样;部分种间的小孢子形态相似,但可通过纹饰类型、裂缝的曲直进行区分。(4)琼海卷柏的小孢子具有独特的网状纹饰,暗示其具有独特的演化途径。(5)卷柏属植物叶表皮的气孔特征、瘤状突起特征,小孢子的颜色、纹饰,与海拔、生境的湿度有一定的相关性,但其形成机理有待进一步研究。     

Monosomic additions in beet (Beta vulgaris) carrying extra chromosomes of B. procumbens

Summary Alien monosomic additions in beet (Beta vulgaris), each carrying one of the nine chromosomes of B. procumbens, were grown in vivo and in vitro to study the effect of the alien chromosomes on plant development. All additional chromosomes caused a reduction of the growth rate in vivo, which, in one case was so strong that some of the plants died as seedlings. In general, the morphological plant characteristics were not very useful to distinguish the addition types; this could have been the results of the wide variation in the recipient parent. However, some developmental characteristics proved to be highly chromosome-specific; for plants in vivo this was annuality, in combination with early or late flowering. If grown in vitro, chromosome specificity was observed for growth type (rosette or elongated stem), occurrence and rate of vitrification, occurrence and morphology of wound callus, formation of additional meristems on the midribs of leaves, formation of roots and a specific reaction to benzylaminopurine (BAP) the medium. Two chromosome types of B. procumbens caused resistance to the beet cyst nematode.     

Importance of secondary habitats and need for ecological management for the conservation of Diphasiastrum tristachyum (Lycopodiaceae, Pteridophyta) in the Vosges Mountains (France)

Diphasiastrum tristachyum, which is a rare Pteridophyte under national protection in France, has been discovered in several places in the Vosges Mountains during the last two decades. With its 20 currently existing populations in these mountains, it appears to be the Diphasiastrum species with the largest current distribution in the Vosges. This species mainly occurs on Vosgian sandstone substrate, in northern or northeastern exposures up to 600 m a.s.l., corresponding to the lower and middle montane levels in the Vosges Mountains. The main habitat (16 sites) are forest track borders. Only in single cases could D. tristachyum be found in other habitats such as a skiing track, a heath submitted to regular fire, an abandoned quarry and a limit of a forest plantation weeded with herbicides. All of these sites are secondary places of young anthropic origin. No really natural and primary habitats of D. tristachyum in the Vosges Mountains could be found. All localities show fragments or beginnings of heath communities belonging to the Genistion alliance. Because of the progressive succession of these communities towards forests if no human regulation occurs, D. tristachyum may have only ephemeral existence in such secondary Vosgian sites. This shows the need for a conservation and management programme which comprises measures like keeping open growing sites by the removal of invading trees, controlled pruning of dwarf shrubs and removal of the soil vegetation and the humus layer. A long-term stabilisation of the Vosgian populations of D. tristachyum may only be possible in this way.     

Heterostyly in species ofNarcissus (Amaryllidaceae) andHugonia (Linaceae) and other disputed cases

The hypothesis ofHenriques andFernandes that several Iberian species ofNarcissus (Amaryllidaceae) are tristylous is reconsidered. Contrary to the opinion ofBateman and most subsequent authors, we believe that the available evidence indicates that some populations ofN. triandrus andN. fernandesii, at least, are tristylous; other populations ofN. triandrus are distylous.Hugonia cf.penicillanthemum (Linaceae) from new Caledonia is distylous, but it remains possible that other species ofHugonia are tristylous. The disputed occurrence of heterostyly in S. African species ofBauhinia (Leguminosae),Cleome (Capparaceae) andAneilema (Commelinaceae), and inAgelaea (Connaraceae) is discussed.     

Comparative karyomorphology and interrelationships ofSelaginella in Japan

Karyomorphological comparisons were made of 16 native and cultivated species ofSelaginella in Japan. The somatic chromosome numbers are 2n=16 inS. boninensis; 2n=18 inS. doederleinii, S. helvetica, S. limbata, S. lutchuensis, S. nipponica, S. selaginoides, S. tama-montana, andS. uncinata; 2n=20 inS. biformis, S. involvens, S. moellendorffii, S. remotifolia, andS. tamariscina; 2n=30 inS. rossii; and 2n=32 inS. heterostachys. The interphase nuclei of all species examined are uniformly assigned to the simple chromocenter type. The metaphase karyotype of 2n=16 (x=8) is 8 m (=median centromeric chromosomes)+8(st+t)(=subterminal and terminal). The group of the species having 2n=18 (x=9) is heterogeneous karyomorphologically: The karyotype ofS. nipponica is 2n=18=6 m+12(st+t),S. tama-montana 10 m+2 sm(=submedian)+6(st+t), andS. uncinata 6 m+7 sm+5(st+t). Although the remaining five species have the common karyotype 8 m+4 sm+6(st+t), the values of mean chromosome length are variable. Another group of the specles having 2n=20 (x=10) is homogeneous, since all species have the same karyotypes 8 m+4 sm+8(st+t) and have similar chromosome size. The karyotype of 2n=30 is 12 m+6 sm+12(st+t) and is suggested to be a triploid of x=10, and 2n=32=16m+16(st+t), a tetraploid of x=8. Thus, three kinds of basic chromosome numbers, x=8, 9, 10 are present in JapaneseSelaginella examined, and their karyomorphological relationships are discussed.