Spore abortion index (SAI) as a promising tool of evaluation of spore fitness in ferns: an insight into sexual and apomictic species

Ferns reproduce through small and usually haploid spores. The general paradigm states that whereas species produce good shaped spores, hybrids are sterile and form aborted spores. Apomictic fern species represent an unusual case, and it is believed that they produce an unbalanced spore spectrum. Until now, no comprehensive comparison of sexual and apomictic taxa using extensive spore fitness data has been published. Based on a representative data set of 109 plants from 23 fern taxa, we accomplished the first robust analysis of spore fitness using spore abortion index (SAI), the ratio of aborted to all examined spores. One thousand spores were analyzed for each plant. Focusing mainly on two major European fern taxa (Asplenium, Dryopteris), we compared this trait for different fern reproductive types (sexual/apomicts/hybrids) and ploidy levels (diploid versus polyploid). Our results confirmed the general assumption that shows higher SAI for apomictic taxa (18%) when compared to sexual taxa (3%). Furthermore, hybrids are characterized by having almost all spores aborted (99.8%) with the notable exception of pentaploid Dryopteris × critica (93.1%), the hybrid between sexual and apomictic taxa. We found no significant difference in SAI between sexual taxa of various ploidy levels or between sexual taxa of genera Dryopteris and Asplenium. Additionally, we carried out an optimization of the SAI method, outlying important guidelines for the use of this method in the future.     

Davallia (Polypodiales: Davalliaceae) macrofossils from Early Miocene Otago (New Zealand) with in situ spores

Fossil fern fronds referable to the extant fern genus Davallia (Polypodiales: Davalliaceae) bearing sporangia with in situ spores are described from the Early Miocene Foulden Maar diatomite deposit, Otago, New Zealand. The fronds are the first published Southern Hemisphere macrofossil record for the family and provide valuable palaeoclimate data supporting warm conditions in Early Miocene New Zealand. The matching of Davallia fronds to the form spore taxon Polypodiisporites radiatus shows that the genus has had a long, apparently continuous history throughout late Cenozoic New Zealand.     

团扇蕨孢子发生和发育的显微观察

利用光学显微镜对膜蕨科（Hymenophyllaceae）团扇蕨(Gonocormus minutus(Blume) Bosch)孢子的发生和发育进行了观察。研究结果表明：团扇蕨孢子为多边圆形,三裂缝不明显,外壁表面光滑,周壁薄,紧贴外壁表面,由周壁形成乳头状或颗粒状纹饰。在外壁形成后期,孢子表面和囊腔中出现大量小球;在周壁形成时期,孢子表面和周围出现较多小球体;小球和小球体参与孢子壁的形成。团扇蕨绒毡层为混合型,内层为周原质团绒毡层;外层为腺质型绒毡层。本文为膜蕨科系统演化和发育生物学研究提供依据。     

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.     

Systematic implications of flavonoid pigments in the fern genus hemionitis (adiantiaceae)

The closely related fern generaHemionitis L. andGymnopteris Bernhardi are separated primarily on differences in leaf architecture and venation. Studies indicate that these characters are highly variable and unreliably diagnostic. Further, the type species of the two genera readily hybridize with each other. Spore morphology, as exhibited by SEM, does not support the traditional alignment of the species in these two genera: some species ofHemionitis andGymnopteris have the same rugose to papillate spores, while other species from both genera possess crested spores. The flavonoid chemistry of these taxa coincides with spore type, i.e., taxa from both genera which possess crested spores produce kaempferol and quercetin 3-0-glycosides, while species with tuberculate spores produce only quercetin 3,4′-0-glycosides. The spore and chemical data suggest a realignment of these taxa within a single genus, which would avoid the rather tenuous dependence on a single vegetative character for generic distinctions.     

Modeling mycorrhizal fungi dispersal by the mycophagous swamp wallaby (Wallabia bicolor)

Despite the importance of mammal‐fungal interactions, tools to estimate the mammal‐assisted dispersal distances of fungi are lacking. Many mammals actively consume fungal fruiting bodies, the spores of which remain viable after passage through their digestive tract. Many of these fungi form symbiotic relationships with trees and provide an array of other key ecosystem functions. We present a flexible, general model to predict the distance a mycophagous mammal would disperse fungal spores. We modeled the probability of spore dispersal by combining animal movement data from GPS telemetry with data on spore gut‐retention time. We test this model using an exemplar generalist mycophagist, the swamp wallaby (Wallabia bicolor). We show that swamp wallabies disperse fungal spores hundreds of meters—and occasionally up to 1,265 m—from the point of consumption, distances that are ecologically significant for many mycorrhizal fungi. In addition to highlighting the ecological importance of swamp wallabies as dispersers of mycorrhizal fungi in eastern Australia, our simple modeling approach provides a novel and effective way of empirically describing spore dispersal by a mycophagous animal. This approach is applicable to the study of other animal‐fungi interactions in other ecosystems.     

Sporangium Exposure and Spore Release in the Peruvian Maidenhair Fern (Adiantum peruvianum,Pteridaceae)

We investigated the different processes involved in spore liberation in the polypod fern Adiantum peruvianum (Pteridaceae). Sporangia are being produced on the undersides of so-called false indusia, which are situated at the abaxial surface of the pinnule margins, and become exposed by a desiccation-induced movement of these pinnule flaps. The complex folding kinematics and functional morphology of false indusia are being described, and we discuss scenarios of movement initiation and passive hydraulic actuation of these structures. High-speed cinematography allowed for analyses of fast sporangium motion and for tracking ejected spores. Separation and liberation of spores from the sporangia are induced by relaxation of the annulus (the ‘throwing arm’ of the sporangium catapult) and conservation of momentum generated during this process, which leads to sporangium bouncing. The ultra-lightweight spores travel through air with a maximum velocity of ~5 m s^-1, and a launch acceleration of ~6300g is measured. In some cases, the whole sporangium, or parts of it, together with contained spores break away from the false indusium and are shed as a whole. Also, spores can stick together and form spore clumps. Both findings are discussed in the context of wind dispersal.     

Ultrastructural study of the microsporidian chytridiopsis typographi (Chytridiopsida: Microspora) infecting the bark beetle,Ips typographus (Scolytidae: Coleoptera), with new data on spore dimorphism

Two types of sporogony of the microsporidian Chytridiopsis typographi in the midgut of adult bark beetle, Ips typographus, have been examined by means of light and electron microscopy. New data are reported on spore dimorphism and on the formation of pansporoblasts in two types of sporogony. Thin-walled spores, larger in size, are formed in a parasitophorous vacuole in the host columnar cells. Thick-walled spores are formed in a minimal vacuole in the host. The ultrastructure of the spore walls and the cyst wall are different from the organization in other microsporidia. Both spore types have identical internal structures and viable spores.     

Biochemistry of fern spore germination: protease activity in ostrich fern spores

Protease activities were detected in quiescent and germinating spores of the ostrich fern (Matteuccia struthiopteris [L.] Todaro). Peak endopeptidase, aminopeptidase, and carboxypeptidase activities were detected 12 to 24 hours after spores began imbibing under light. There was a correlation between activities of proteases, the onset of a decline in levels of soluble protein, and an increase in levels of free amino acids. The earliest visible event of spore germination, breakage of the spore coat and protrusion of a rhizoid cell, was observed after peak protease activity, 48 to 72 hours after the start of imbibition. Results of this study demonstrate similarities in the pattern of protease activities during germination of ostrich fern spores to those of some seeds.     

Effect of tree species on the coverage and field persistence of Bacillus thuringiensis spores

The effect of leaves of Quercus agrifolia, Cercis occidentalis, Eucalyptus globulus, and Juglans regia on the initial deposit and subsequent rate of decay of viable spores of Bacillus thuringiensis was investigated. Significant differences in the size of the initial viable spore deposit were found between the various species, with E. globulus markedly lower than the others. The viable spore decay patterns of the various species were analyzed using a segmented linear model and significant differences in initial rates of decay were also measured. Thus it appears that the characteristics of the leaves of the treated plants may have an effect on the coverage and field persistence of viable B. thuringiensis spores. Some possible mechanisms for these effects are discussed.     

Fern spore and pollen airspora profile of Singapore

Monitoring atmospheric fern spore and pollen loads in Singapore was initiated in June 1990. Aside from the more numerous fungal spores, fern spores and pollen grains made up 6.2–8.6% and 4.4–5.4% of the total airspora sampled, respectively. The most frequently encountered fern spores, in descending order, were those of Nephrolepis auriculata, Dicranopteris linearis, Stenochlaena palustris, Asplenium nidus, Pteridium aquilinum, and Dicranopteris curranii. For pollen grains, the most frequently encountered, in descending order, were Elaeis guineensis, Casuarina equisetifolia, Acacia auriculiformis, Kyllingia polyphylla, Podocarpus, and Poaceae pollen grains. Seasonal patterns for individual fern spore or pollen types were discernible despite the relatively uniform tropical climate in Singapore. The fern spore and pollen calendar for the period 1991–1995 was compared to that of 2005–2006 as a follow-up study to keep abreast with the rapidly changing landscape of Singapore. Diurnal patterns showing a late morning to afternoon peak period were seen in fern spores, while the peak period was in the morning for pollen types studied with the exception to oil palm pollen. Additionally, association between fern spore and pollen counts and local meteorological conditions were also analyzed and found to be highly correlated. This study has thus identified the fern spores and pollen airspora components, and determined the calendars, as well as diurnal profiles of the Singapore airspora and provides invaluable information for allergy studies by highlighting the trigger sources present in the environment.     

Fern Spore Longevity in Saline Water: Can Sea Bottom Sediments Maintain a Viable Spore Bank?

Freshwater and marine sediments often harbor reservoirs of plant diaspores, from which germination and establishment may occur whenever the sediment falls dry. Therewith, they form valuable records of historical inter- and intraspecific diversity, and are increasingly exploited to facilitate diversity establishment in new or restored nature areas. Yet, while ferns may constitute a considerable part of a vegetation’s diversity and sediments are known to contain fern spores, little is known about their longevity, which may suffer from inundation and - in sea bottoms - salt stress. We tested the potential of ferns to establish from a sea or lake bottom, using experimental studies on spore survival and gametophyte formation, as well as a spore bank analysis on sediments from a former Dutch inland sea. Our experimental results revealed clear differences among species. For Asplenium scolopendrium and Gymnocarpium dryopteris, spore germination was not affected by inundated storage alone, but decreased with rising salt concentrations. In contrast, for Asplenium trichomanes subsp. quadrivalens germination decreased following inundation, but not in response to salt. Germination rates decreased with time of storage in saline water. Smaller and less viable gametophytes were produced when saline storage lasted for a year. Effects on germination and gametophyte development clearly differed among genotypes of A. scolopendrium. Spore bank analyses detected no viable spores in marine sediment layers. Only two very small gametophytes (identified as Thelypteris palustris via DNA barcoding) emerged from freshwater sediments. Both died before maturation. We conclude that marine, and likely even freshwater sediments, will generally be of little value for long-term storage of fern diversity. The development of any fern vegetation on a former sea floor will depend heavily on the deposition of spores onto the drained land by natural or artificial means of dispersal.     

Sporoderm ultrastructure ofLophosoria andCyatheacidites (Filicopsida): Systematic and evolutionary implications

The distinctive spores produced byLophosoria, an extant monotypic fern, are examined ultrastructurally and correlated with the sporoderm of fossilCyatheacidites. The morphological and ultrastructural similarity of the two taxa provide additional information that can be used to trace the evolutionary history of this spore type.     

Effect of slug mycophagy on Tuber aestivum spores

Truffles in the genus Tuber produce subterranean fruiting bodies that are not able to actively discharge their spores in the environment. For this reason, truffles depend on mycophagous animals for reproduction. Fungus consumption (mycophagy) is a behaviour typical of both vertebrates and invertebrates. Mammals, especially rodents, are the most studied group of mycophagists and have been found to consume a great variety of fungi. Among invertebrates, mycophagy is documented in arthropods, but rarely in molluscs. In our study we assessed the effect on the morphology and mycorrhizal colonization of Tuber aestivum spores after passage through the gut of slugs (Deroceras invadens) and, for comparison, of a house mouse (Mus musculus). Light, scanning electron and atomic force microscopy revealed that the digestion, especially by slugs, freed spores from the asci and modified their morphology. These are believed to be the reasons why we observed an improvement in oak mycorrhization with the slug and rodent ingested spores in comparison to a fresh spore inoculation. We also demonstrated by molecular barcoding that slugs’ guts sampled on a Tuber melanosporum truffle ground contain spores from this species and Tuber brumale, further suggesting that some invertebrates are efficient Tuber spore dispersers.     

Micrograzers and spore release in Iridaea laminarioides Bory (Rhodophyta: Gigartinales)

Preferential consumption of algal reproductive tissues has been predicted to potentially reduce algal fitness by reduction of reproductive output. This study shows that the amphipod Hyale media (Dana) has a marked trophic preference for mature cystocarpic tissues of Iridaea laminarioides Bory. However, during the feeding process the amphipod tears the cystocarps releasing large numbers of spores into the water column. Germination and growth rate experiments indicate that grazer-mediated release does not affect further spore development. Field populations of I. laminarioides might have unopened cystocarps even in senescent, decaying fronds, while the total number of open cystocarps in mature and senescent fronds increases at higher amphipod density sites, suggesting a facilitation mechanism. Laboratory observations indicate spores can stick to the legs and body sides of the amphipods while a fraction of the ingested spores survives passage through the amphipod digestive tract. Thus, the amphipod seems ecologically important not only as a grazer but also as spore releaser and disperser.     

Fatty acid composition of fern spore lipids

Fatty acid composition of lipids isolated from spores of different fern groups show differences between the families whereas species variations within the families are smaller. As in seed fats, the spore lipids are mainly triglycerides, with the exception of Osmunda where free fatty acids accumulate. The spore lipids contain as major components oleic, linoleic, and palmitic acid although those of the sporophylls contain C-20 polyunsaturated acids.     

Reduction of consumption by grasshoppers (Orthoptera: Acrididae) infected with Nosema locustae canning (Microsporida: Nosematidae)

The effect of ingestion of Nosema locustae Canning spores on feeding by grasshoppers was measured in simultaneous laboratory and field experiments. After 21 days, fourth-instar Melanoplus sanguinipes (F.) nymphs, administered spores at the rates of 0, 2.0 × 10⁴, 2.0 × 10⁵, and 2.0 × 10⁶ per grasshopper, showed dry matter consumption of 102, 87, 64, and 26 mg in 48 hr, respectively. Rate of inoculation was a significant factor in suppression of feeding after correction for the effects of developmental stage, sex, and body weight. The quantity of dry matter consumed decreased linearly with increasing rate of spore ingestion. Experiments on 50 caged 1-m² plots on pasture grass yielded similar trends in per capita consumption independent of the effects of mortality. Field consumption per integrated grasshopper-day was 108, 77, 31, and 27 mg dry wt at the four inoculation rates, over 20 days.     

Fern and lycopod spores rain in a cloud forest of Hidalgo,Mexico

The aim of this study was to determine the composition of the “spore rain” of ferns and lycopods in a cloud forest. We tested whether the canopy impedes spore dispersal to surrounding areas and how spore dispersal is affected by rainfall. The spores were captured with a modified Bush–Gosling trap placed at 30 cm above ground level in forested and non-forested sites from March 2009 to February 2010. We collected 2462 fern spores from 158 morphospecies of which 76 were identified to species level. Thirty-seven species were found exclusively in the spore rain, and 39 were found as sporophytes as well (local component). Mean daily spore density (spores m^?2) was calculated to find the sporulation period for each species. Twenty species showed seasonal patterns of sporulation. The highest spore density was found at the forested site (70 morphospecies and 1856 spores), of which 39 morphospecies (1482 spores) corresponded to the local vegetation. Fifty-five taxa were shared between the forested and non-forested site. In the non-forested site, 605 spores were captured belonging to 64 species. The density of spore rain between sites was significantly different. The rainfall amount was the same at both sites, with a dry period in March, April, and July 2009, and February 2010. There was a negative effect of rainfall on spore rain. The main sporulation occurred in the dry season with strong winds. Although the canopy inhibits airborne dispersal of fern spores, a small amount of spores can disperse beyond the canopy and reach surrounding areas. The rainfall might wash spores to ground and favor the colonization and the establishment of new populations.     

Mass production and storage of Vairimorpha necatrix (protozoa: Microsporida)

Mass production and storage methods were evaluated for maximization of spores of Vairimorpha necatrix, a promising protozoan for microbial control due to its virulence and prolificity in lepidopterous pests. In vivo spore production was at a maximum when 3rd instar Heliothis zea were exposed to 6.6 spores/mm² of artificial diet surface and reared for 15 days. Approximately 1.67 × 10¹⁰ spores/larva were produced, or ca. 1 × 10¹⁰ spores/larva after partial purification of the spores by homogenization of the larvae in water, filtration, and centrifugation. The spores were inactivated by relatively short exposures to several chemicals which were tested to counteract contamination of the diet surface by fungi in the spore inoculum. Spores of V. necatrix were stored at refrigerated and freezing temperatures for up to 2 years and bioassayed periodically with 2nd instar H. zea. Spores lost little infectivity after 23 months at 6°C if they were stored in a purified water suspension plus antibiotic, but they were noninfective after 18 months at 6°C if stored in host tissue. Storage at ?15°C caused little loss of infectivity whether the spores were stored in water and glycerine, in host tissue, or after lyophilization. The spores withstood lyophilization in host cadavers better than in purified water suspension. Samples of a dry V. necatrix-corn meal formulation, which was prepared for field efficacy tests and stored at ?15° and 6°C, were highly infective after 9 months. Large numbers of V. necatrix spores can thus be produced and later made available for microbial control field trials with little loss of infectivity.