Spore morphology of the genus Dryopteris Adans. (Dryopteridaceae) in Korea

Spore morphology of the Korean members of the genus Dryopteris was examined by scanning electron microscopy. In particular, spores of D. hangchowensis, D. woodsiisora, D. saxifragivaria, and D. subexaltata were examined here for the first time. Three perispore types were recognized on the basis of the shape of macro-ornamentations on perispore: these include rugate, echinate, and spinose. The rugate perispore type appeared to be the most common in the genus, and probably represents plesiomorphic condition in the genus Dryopteris. The echinate perispore type is found in D. expansa, and spinose perispore type in D. formosana. The results of this study demonstrate that the perispore characteristics, particularly the shape of macro-ornamentation and surface texture, have diagnostic value at the species level. Indeed, most of the Dryopteris taxa in Korea could be distinguished by their perispore characters. In addition, spores of sexual diploid individuals of D. saxifraga were much smaller than those of agamosporous triploid individuals, supporting the contention that the size of spores in ferns is generally related to ploidy levels.     

水蕨孢子壁的形成和发育

利用光镜、扫描电镜和透射电镜对水蕨科（Parkeriaceae）水蕨（Ceratopteris thalictroides（L．）Brongn．）孢子壁的形成和发育进行了研究。结果表明,水蕨孢子呈辐射对称,三裂缝,表面具肋条状纹饰。孢子壁由内壁、外壁和周壁三部分构成。在四分体阶段外壁已基本形成,其外壁显著,表面光滑,质地均匀,由孢粉素形成,外壁厚约3—5μm,脊高约5—7μm。周壁由绒毡层残余物在外壁表面沉积形成,较薄,厚度只有0．1μm,表面具有杆状突起。研究结果对揭示孢子纹饰和孢子壁各层的形成过程、来源和稳定性有一定的意义,并为蕨类植物孢粉学和系统学研究提供基础资料。     

水蕨孢子壁的形成和发育

利用光镜、扫描电镜和透射电镜对水蕨科(Parkeriaceae)水蕨(Ceratopteris thalictroides (L.) Brongn.)孢子壁的形成和发育进行了研究。结果表明, 水蕨孢子呈辐射对称, 三裂缝, 表面具肋条状纹饰。孢子壁由内壁、外壁和周壁三部分构成。在四分体阶段外壁已基本形成, 其外壁显著, 表面光滑, 质地均匀, 由孢粉素形成, 外壁厚约3-5 μm, 脊高约5-7 μm。周壁由绒毡层残余物在外壁表面沉积形成, 较薄, 厚度只有0.1 μm, 表面具有杆状突起。研究结果对揭示孢子纹饰和孢子壁各层的形成过程、来源和稳定性有一定的意义, 并为蕨类植物孢粉学和系统学研究提供基础资料。     

Spore wall ultrastructure of Polypodiaceae from north-western Argentina

The spore wall ultrastructure of Campyloneurum, Microgramma, Pecluma, Phlebodium, Pleopeltis and Serpocaulum (Polypodiaceae) from north-western Argentina has been studied using transmission electron microscopy (TEM). The exospore is 0.4–3 μm thick, two-layered and variously ornamented in all taxa. The exospore surface is distinctive, but in general ultrastructure the exospore is similar in all species studied. The structural elements of the exospore consist of cavities in the inner part as well as channels with a radial orientation and channels at both sides of the laesura. Variation in the exospore surface was observed in spores at different stages of maturation. The perispore is darkly contrasted and 0.04–2 μm thick. Three different structure types were recognised, including fibrillar, multilamellar and lacunose. Scattered globules and spherules were always present on the perispore surface. The structural variability of the perispore was surveyed within complete sporangia. We concluded that the observed variability may be related to the stage in spore maturation and, consequently, to the stages in perispore differentiation. As the exospore ultrastructure is similar and interpreted as related to functional activity in the studied material, it cannot be used for systematic delimitations at this generic or specific level.     

Spores in situ and problems of the classification of Mesozoic tree ferns

Although tree ferns dominated the Mesozoic flora, their taxonomic relationships are poorly understood at the generic level, and next to nothing can be said of evolutionary trends within the group. At least five genera are recognized based on the remains of spore-bearing structures. However, the dispersed spores belong to the same generalized morphotype, and cannot be assigned to genera based on macroscopic remains of fertile leaves. Electron microscopy of spores in situ may partly resolve these problems providing additional criteria for classification of spore-bearing structures and disperse spores. We studied in situ spores of the Early Cretaceous Alsophilites nipponensis (Oishi) Krassilov, which are comparable to dispersed spores Cyathidites minor Coup. Spore wall micromorphology and ultrastructure indicate their affinities with the modern genus Alsophila R. Brown. Only occasional poorly preserved striate sculptures survive the standard treatment of maceration of the perispore. Our data confirm the primitive status of the species with a great number of spores per sporangium, thick unsculptured exospore consisting of two ultrastructural layers, and the possibility that whole sporangia with unshed spores can function as dispersal units.     

Spore morphology and wall ultrastructure of Hymenophyllaceae Link (Pteridophyta) from north-west Argentina

The family Hymenophyllaceae is represented in the study area by six species in two genera, Hymenophyllum J. E. Smith and Trichomanes L. The study was based on herbarium material and spores were studied under light microscope (LM), scanning electron microscope (SEM) and transmission electron microscope (TEM). Both genera have trilete spores, 23 to 45 μm in equatorial diameter, with an ornamentation of echinulae and cones in Hymenophyllum and of verrucae, gemmae and granules in Trichomanes. Mature spores have a sporoderm composed of a perispore, an exospore and a fibrillar endospore; the exospore is 0.5 to 2.5 μm thick, compact and with an irregular margin. In some cases radial channels and other channels associated with the middle and inner parts of the laesurae were evident. A series of cavities filled with an opaque content line the inner margin of the exospore. The perispore is 20 to 400 nm thick and unevenly differentiated along the surface of a same spore. Under TEM, two main differentially contrasted portions could be distinguished: a dark massive portion with structural components could not be distinguished, and a light portion with several plates arranged in piles. The inner surface of the perispore exhibit short scales. Globules are immersed within the perispore at some depth from the perispore surface and others connected to it by structural threads. The spore characters observed including shape, ornamentation, laesurae length and wall structure are useful in distinguishing the two genera studied, but less useful in differentiation at the species level.     

The spores of Pteris cretica complex (Pteridaceae-Pteridophyta) in America

Spore morphology in the Pteris cretica complex, including Pteris brasiliensis, P. ciliaris, P. cretica, P. denticulata var. denticulata, P. denticulata var. tristicula, P. ensiformis, P. multifida and P. mutilata was examined using light and scanning electron microscope. The spores are trilete, triangular to circular in shape, with an equatorial thickening (= cingulum). Equatorial diameter ranges from 29 to 69 μm, and polar diameter from 18 to 53 μm. Exospore is proximally verrucate and distally rugate, with the exception of Pteris ensiformis, which has cones on both polar faces. Perispore is generally less than 1 μm thick, apparently single-layered in section, and translucent under light microscope. Spheroids were frequently observed on the surfaces of both perispore and exospore. Hyaline spores lacking cingulum were found in all specimens. Pteris brasiliensis, P. cretica and P. denticulata exhibit pronounced polymorphism and, in addition to trilete spores these taxa also produce atypical spores such as tetralete, monolete and intermediate types.     

Spore morphology of the north Asian members of Cystopteridaceae

Spores of Cystopteridaceae from northern Asia were examined using scanning electron microscopy. To evaluate the utility of spore morphology in the taxonomy of each genus, we examined spores of 14 species: seven species each of Gymnocarpium and Cystopteris. Among these are 12 species occurring in northern Asia and two species from other regions for comparative studies. The study focused particularly on perispore characters and spore size. Spores of all species examined are monolete, bean-shaped, with a range in spore size of 26–56 × 18–37 μm for Cystopteris and 25–48 × 16–34 μm for Gymnocarpium. The perispore is morphologically diverse within Cystopteris, but less so within Gymnocarpium. The perispore of the Cystopteris spores is characterised by folds and spines that are separate or form complex sculptural elements. Sacci, ridges and flanges, sometimes on the same spore, are characteristic of the perispore of Gymnocarpium. Spores have straight laesura over which the perispore forms a crest. The crest represents a high and flat fold, which is entire, foveolate or reticulate.     

Abnormal spore morphology and wall ultrastructure in Anemia tomentosa var. anthriscifolia and A. tomentosa var. tomentosa (Anemiaceae)

The spores of Anemia tomentosa var. anthriscifolia and A. tomentosa var. tomentosa were studied focusing the attention on their abnormalities. The study was based on fresh and herbarium material and the spores were examined with light, scanning and transmission electron microscopy. Normal, abnormal and abortive spores were observed in both taxa. The normal spores were trilete, triangular in polar view, and the ornamentation consisted of parallel ridges separated by narrow and smooth grooves. The spores were observed in monads, dyads, triads and tetrads. The abnormal spores were monolete, trilete, tetralete or alete with great variations in size. In fact, some spores were almost double the size of the normal ones. Some differences were also found in the ornamentation of the spores. Aborted and not completely developed spores were also observed in the specimens. The wall ultrastructure of the taxa was studied for the first time. The exospore was two-layered with numerous cavities inside its structure, and the perispore was also two-layered. The results revealed that the sporoderm ultrastructure of both normal and abnormal spores of the taxa analyzed was very similar.     

Unique expression of a sporophytic character on the gametophytes of notholaenid ferns (Pteridaceae)

? Premise of the study: Not all ferns grow in moist, shaded habitats; some lineages thrive in exposed, seasonally dry environments. Notholaenids are a clade of xeric-adapted ferns commonly characterized by the presence of a waxy exudate, called farina, on the undersides of their leaves. Although some other lineages of cheilanthoid ferns also have farinose sporophytes, previous studies suggested that notholaenids are unique in also producing farina on their gametophytes. For this reason, consistent farina expression across life cycle phases has been proposed as a potential synapomorphy for the genus Notholaena. Recent phylogenetic studies have shown two species with nonfarinose sporophytes to be nested within Notholaena, with a third nonfarinose species well supported as sister to all other notholaenids. This finding raises the question: are the gametophytes of these three species farinose like those of their close relatives, or are they glabrous, consistent with their sporophytes? ? Methods: We sowed spores of a diversity of cheilanthoid ferns onto culture media to observe and document whether their gametophytes produced farina. To place these species within a phylogenetic context, we extracted genomic DNA, then amplified and sequenced three plastid loci. The aligned data were analyzed using maximum likelihood to generate a phylogenetic tree. ? Key results: Here we show that notholaenids lacking sporophytic farina also lack farina in the gametophytic phase, and notholaenids with sporophytic farina always display gametophytic farina (with a single exception). Outgroup taxa never displayed gametophytic farina, regardless of whether they displayed farina on their sporophytes. ? Conclusions: Notholaenids are unique among ferns in consistently expressing farina across both phases of the life cycle.     

MATERNAL INHERITANCE OF THE CHLOROPLAST AND MITOCHONDRIAL GENOMES IN CHEILANTHOID FERNS

Molecular data from the chloroplast genome are being used to reconstruct the phylogeny and revise the problematic taxonomy of the xerically adapted cheilanthoid ferns. Chloroplast DNA based phylogenies trace maternal, paternal, or biparental lineages, depending on the mode of inheritance of the chloroplast genome, and instances of all three modes of inheritance are known in the seed plants. Evidence for biparental and uniparental inheritance in ferns has been presented, but the distinction between maternal and paternal uniparental inheritance has not been rigorously made, and the mode of inheritance in cheilanthoid ferns is completely unknown. Based on a natural hybrid population in the cheilanthoid genus Pellaea in which the maternal and paternal derivations of the hybrid are unambiguously known, restriction fragment length polymorphisms of chloroplast DNA demonstrated simple maternal inheritance of the chloroplast genome. This hybrid complex was also examined for restriction fragment length polymorphisms of its mitochondrial DNA, providing the first direct evidence that the mitochondrial genome in ferns is maternally inherited.     

华中铁角蕨孢子纹饰形成的超微结构观察

利用透射电子显微镜对铁角蕨科(Aspleniaceae)华中铁角蕨(Asplenium sarelii Hook.)孢子及其纹饰的形成过程进行观察。结果表明:①华中铁角蕨孢子囊发育为薄囊蕨型;②孢子外壁表面光滑,远极面的外壁厚约0.8~1.1μm,近极面的外壁厚约1.4~1.8μm;③孢子周壁厚度约4~5μm,染色较外壁深,分为内层和外层;内层紧帖外壁表面,其上具柱状、瘤状或疣状突起;外层向外隆起形成脊状纹饰的轮廓,脊的下方具空腔,脊的顶端具翅;④铁角蕨型与鳞毛蕨型孢子外壁和周壁纹饰的形成过程具有相似性;⑤孢子的成熟度对于孢子形态的研究是至关重要的,只有完全成熟的孢子的表面纹饰才是稳定的。     

Ultrastructural and molecular characterization of cyanobacterial symbionts in Dictyocoryne profunda (polycystine radiolaria)

Cyanobacterial symbionts were detected in the extracytoplasm of the polycystine radiolarian Dictyocoryne profunda Ehrenberg. The bacterial symbionts were observed as numerous spherical bodies ~0.5?C1.0???m in diameter under transmission electron microscopy. They were present in a very restricted location close to the periphery of the host radiolarian shell, adjacent to the central capsular wall. Several cells of them may have been in the process of cell division or just divided. The symbionts had thylakoid-like structures, which ran around the cell periphery in two or three concentric layers. Based on the small subunit ribosomal DNA (16S rDNA) phylogenetic analyses, the intracellular symbiotic bacteria grouped with cyanobacteria belonging to the genus Synechococcus. Three sequences, one from each of three specimens of D. profunda, collected in March/October 2009 and March 2010 from the East China Sea, were the same and branched within Synechococcus clade II, that is characterized by strains with low amounts of phycourobilin (PUB).     

Spore morphology and ultrastructure of Cyathea (Cyatheaceae,Pteridophyta) species from southern South America

Spore sculpture and wall structure of eight Cyathea (Cyatheaceae) species from southern South America were studied using light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) techniques. Two layers, i.e. an inner and an outer layer, were observed in the perispore. The inner layer has two strata: the inner stratum is attached to the exospore and composed of rodlets tangentially oriented to the spore surface and randomly intermixed; the outer stratum consists of a three-dimensional network of rodlets with either free or fused distal edges forming spinules. The outer layer is thin, darkly contrasted and covers the rodlets. In most cases, the exospore has two layers and a pitted surface. In Cyathea atrovirens, the exospore surface is smooth, while in C. delgadii and C. myriotricha it is verrucate. The homogeneity of perispore features within the genus Cyathea is evident, while exospore features are heterogeneous. The exospore has different kinds of surface-structures that are of potential interest for assessing evolutionary trends within the group.     

Partial purification and characterization of peroxidases from the leaves of Sapindus mukorossi

Peroxidases were isolated from Sapindus mukorossi (Reetha) and partially purified using acetone precipitation, ion-exchange chromatography with a 14-fold purification, 22% recovery and a specific activity of 266?×?10³ units/mg protein. Sapindus peroxidases (SPases) showed six bands after acetone precipitation and one distinct band after ion exchange chromatography on Native-PAGE after zymography. Enzymes purified by ion exchange chromatography were loaded on Sepahdex G-50 superfine column and their molecular weight was reported to be 25?kDa. They showed temperature optima at 50°C and pH optima at 5.0.?km for SPases was reported to be 1.05?mM and 0.186?mM for guaiacol and H₂O₂ respectively. The V_max/K_m value for o-dianisidine was 449 while for H₂O₂ it was 5?×?10⁵. Protocatechuic acid acts as a potent inhibitor for SPases (6.0% relative activity at 4.5???M) but ferulic acid inhibits its activity at a much lower concentration (0.02???M). Enzymes were stimulated by metal cations like Cu²⁺, Ca²⁺ (145, 168; percentage relative activity respectively) and showed mild inhibition (up to 20%) with Mn²⁺ and Mg²⁺. Alanine stimulated the enzyme activity (up to 33%; at 0?C100???M) while other amino acids like cysteine, methionine, tryptophan and tyrosine inhibited the SPases (13?C57% at 0?C100???M).     

Spore wall ultrastructure in Anogramma species (Pteridaceae) from Argentina

The sporoderm ultrastructure of Anogramma Link species, which grow in Argentina, was studied using scanning and transmission electron microscopy. The species A. chaerophylla (Desv.) Link and A. lorentzii (Hieron.) Diels were studied with TEM for the first time. The spores of both species have a sculptured, apparently two-layered exospore. The perispore is uniformly thickened on the whole surface: in A. chaerophylla it is three-layered, while, in A. lorentzii it is single-layered with a complex structure. Spherules are present on the perispore surface or incorporated into the structure of A. chaerophylla while, globules exist on and within the perispore in A. lorentzii. The sporoderm ultrastructure in these two species was compared with other cingulate genera within the Pteridaceae. The characteristics found in this work, with respect to spore wall structure and general morphology, suggest that these characters may differentiate species within genus.     

Morphology and ultrasctructure of spores Klukia tyganensis Krassilov (Schizaeaceae, Filicales) from the Berriassian of the Tyrma Depression (Russian Far East)

In situ spores of the fern Klukia tyganensis Krassil. from the coal-bearing deposits on the right bank of the Tyrma River (Berriassian, Bureya River Basin, Tyrma Depression) have been examined using light, scanning and transmission electron microscopy. The spores of K. tyganensis are demonstrated to be similar in the ultrastructure of sporoderm to that of some species of the extant genus Anemia and to differ from the spores of Lygodium in the ultrastructure of perispore and exospore.     

Calyptospora empristica n. sp. (Eimeriorina: Calyptosporidae) from the Liver of the Starhead Topminnow,Fundulus notti1

Oocysts of Calyptospora empristica n. sp., the second described species in its genus, are described from the freshwater starhead topminnow, Fundulus notti, in southern Mississippi. Oocysts are 22 μm in diameter with a wall about 20 nm thick and have no residuum, micropyle, or polar granule. Sporocysts are spheroid, 9 × 5 μm with a two-layered wall approximately 120 nm thick. They have an oblong apical opening at the anterior pole, a single ornamented sporopodium approximately 5.7 μm long at the posterior pole, and a residuum. An intermediate host, most likely the freshwater grass shrimp Palaemonetes kadiakensis, is probably required to complete the life cycle.     

Further progress towards the delimitation of <Emphasis Type="Italic">Cheilanthes</Emphasis> (Cheilanthoideae,Pteridaceae), with emphasis on South American species

Cheilanthoid ferns (Cheilanthoideae sensu PPG 1 2016) constitute an important group within the Pteridaceae and are cosmopolitan in distribution. In South America, there are 155 species distributed in 13 genera, among which the largest are Adiantopsis (35), Cheilanthes (27), and Doryopteris (22). Most of the cheilanthoid species are morphologically adapted to grow in arid to semi-arid conditions and show convergent evolution, which has implied difficulties in defining the genera throughout their taxonomic history (Copeland 1947, Tryon & Tryon 1973, Gastony & Rollo 1995, 1998, Kirkpatrick Systematic Botany, 32: 504–518, 2007, Rothfels et al. Taxon, 57: 712–724, 2008). Here, we sequenced two plastid markers (rbcL?+?trnL-F) of 33 South American cheilanthoid species, most of which have not been included in phylogenetic analyses previously. The South American species were analyzed together with South African and Australasian Cheilanthes and representatives of related cheilanthoid genera. The phylogenetic analysis showed that most Cheilanthes species are related to the genus Hemionitis, constituting different groups according to their distribution; moreover, three species—C. hassleri, C. pantanalensis, and C. obducta—appear as the sister clade of Hemionitis. Cheilanthes micropteris, the type species, is strongly supported in a clade with Australasian Cheilanthes plus five South American Cheilanthes species, all of which show a reduction in the number of spores per sporangium; this feature would be a synapomorphy for core Cheilanthes s.s. We found no support uniting other South American Cheilanthes to either the group of South African Cheilanthes or to core Cheilanthes s.s. On the other hand, C. geraniifolia, C. goyazensis, and C. bradei formed a clade related to Doryopteris that, with further study, could be considered as a new genus. The phylogenetic hypotheses presented here contribute substantially to the delimitation of Cheilanthes s.s. and related groups and provide the basis for re-examining the generic taxonomy.