Growth and reproductive characteristics in artificially formed clonal gametophytes ofDryopteris filix-mas (Dryopteridaceae)

Artificially formed clonal gametophytes ofDryopteris filix-mas (L.)Schott were able to grow and reproduce, although growth rates, proportions of hermaphrodites and reproductive efforts were low. Variable density and nutrient levels appeared to affect gametophytic life histories and they continued to influence the viability of the developing sporophyte generation. Differences between populations and sporophytes were discovered in life histories. Hermaphroditic gametophytes were considerably larger and possessed higher viability than did males. Clear differences between clones in the level of phenotypic plasticity in gametophyte size were observed. No neighbor effect on gametophyte sex expression was detected with the densities and nutrient levels used.     

90. K. Jeremias und Elisabeth Frank: Das jahresperiodische Verhalten der Zucker und der Stärke in Wedeln und Rhizomen; von Athyrium filix-femina und Dryopteris dilatata

1. In Wedeln und Rhizomen von Athyrium filix-femina und von Dryopteris dilatata wird der Jahresgang der Zucker und der Stärke bestimmt. 2. Die sommergrünen Wedel von Athyrium besitzen im Juni und im November je ein Zuckermaximum, das von Glucose, Fructose und Saccharose gebildet wird. Ein Zuckerminimum, das mit dem Stärkemaximum zusammenfällt, liegt im Mai. 3. Die Wedel von Dryopteris und die Rhizome beider Arten zeigen ein ausgeprägtes winterliches Zuckermaximum. In den Rhizomen wird während der kalten Jahreszeit Raffinose angehäuft. Der Stärkegehalt ist relativ hoch, nur während der Hauptwachstumsperiode geht er deutlich zurück. 4. Das jahresperiodische Verhalten der Zucker und der Stärke zeigt keine grundsätzlichen Unterschiede zu dem aus den Spermatophyten bekannt gewordenen.     

Fertilization and rejection of spermatozoids by egg cells in artificially pollinated ovules ofEncephalartos (Zamiaceae)

The structure and behaviour of free female, male and proembryonal nuclei ofEncephalartos villosus Lem. were studied during a light-microscopical investigation of serially sectioned archegonia in successfully pollinated ovules. Before spermatozoids were released from the pollen tubes into the archegonial chamber, the ventral canal nucleus had disintegrated in the neck region of the egg cell among minute, amoeboid bodies with PAS-positive granules. In archegonia containing multiple spermatozoids, the egg nucleus was unobtrusive and syngamy followed by proembryo formation regularly resulted. The egg cell usually reacted violently in archegonia penetrated by a single spermatozoid. These reactions were regarded as rejection phenomena and considered as indicators that the egg cell can differentiate between compatible and incompatible male gametes.     

Fern rhizomes as fodder in Norway

Background

Although ferns are often known under collective names in Norway, e.g. blom, a substantial number of vernacular names for individual fern species are known, in particular for useful or poisonous taxa. In the past, the rhizomes (Norwegian: moldfôr) of selected species were collected for fodder. Only scattered records of such use are available from southern Norway, and the tradition’s core area is found in the two North Norwegian counties of Nordland and Troms, in accordance with the longer winters encountered in the north, frequently leading to fodder shortage in early spring. The tradition extends northeastwards into Finnmark, but is less well documented there. Although numerous sources mention the use of fern rhizomes for fodder, the fern species hiding behind the tradition are incompletely known. This paper aims at reviewing available data in terms of identifyng the species used for fodder, the history and geographical distribution of such use, and other relevant traditions, e.g. the timing and mode of collection, and the way the rhizomes were used.

Methods

The study is based on data extracted from a variety of archival and literature sources; the latter retrived from my database of more than 7500 publications providing information on plant names and plant uses in Norway.

Results

More than 200 individual records mention the use of fern rhizomes for fodder in Norway. Only a fraction of these, typically made by botanist recording data on plant uses, provides information on the identity of the species used. Based on these, Dryopteris filix-mas and Matteuccia struthiopteris stand out as the most important species serving as sources of fern rhizomes for fodder. Locally, Dryopteris expansa was the preferred species, and this taxon may to some extent be overlooked in the records so far available. With a few exceptions, Norwegian folk tradition singles out Athyrium filix-femina as a harmful and poisonous species, causing livestock to go blind and lame, but whether this is true or not, remains unknown; the symptoms are in fact documented elsewhere as a consequence of poisoning due to Dryopteris filix-mas. In coastal north Norway, fern rhizomes were regularly collected for fodder, both in late autumn and early spring, and used to remedy a recurrent shortage of fodder in late winter and spring. Locally, the tradition of collecting fern rhizomes lived on until the 1940’s or 1950’s. Although mainly a tradition of the ethnic Norwegians, it had also been adopted by the farmers belonging to the Finnish and Sámi ethnic minorities.

Conclusion

Fern rhizomes have a long tradition as an additional fodder for livestock in Norway. Preferred species were Matteuccia struthiopteris and Dryopteris filix-mas, locally also Dryopteris expansa. Athyrium filix-femina was considered to be poisonous, and usually avoided.

     

Arbuscular mycorrhizas on Athyrium yokoscense and A. niponicum grown at a lead-contaminated site

To investigate the occurrence of arbuscular mycorrhizas (AM) and their morphological types, four Athyrium yokoscense and five A. niponicum individuals were collected at a lead-contaminated site. Three of the A. yokoscense and four of the A. niponicum formed AM structures. However, the percentage of AM formation within both species differed significantly among individuals. Morphological types of most AM structures were the Paris-type, except for one A. yokoscense that formed both Paris-type and Arum-type-like structures in different parts of the same root section. These results demonstrate that the two Athyrium species form AM associations in lead-contaminated soil.     

Bestimmung des Schleimgehaltes myxospermer Diasporen verschiedener Angiospermenfamilien

The mucilage content of structurally differing myxospermatic diaspores from 49 species belonging to 19 families ofAngiospermae has been determined by applying various extraction procedures. The results demonstrate no obvious relationship between the size, mucilage quantity, and the swelling factor of the diaspores studied. Furthermore, mucilage producing structures and structural peculiarities of the mucilages themselves are elaborated.

     

<Emphasis Type="Italic">Athyrium crassicaule</Emphasis> (<Emphasis Type="Italic">Woodsiaceae</Emphasis>), a new fern species from the Drakensberg,southern Africa

Summary  A new Athyrium species, A. crassicaule J. P. Roux (Woodsiaceae), is described from the Drakensberg in Lesotho and South Africa. A key to the Athyrium species in Lesotho, South Africa and Swaziland is provided.     

Biosystematic investigations on the lady fern(Athyrium filix-femina)

In EuropeAthyrium filix-femina has a constant chromosome number (2n = 80) and is sexual. The normal type of reproduction is intergametophytic crossing. In the gametophyte phase there is a hormone system which induces dark germination of spores and antheridium formation. Sporophytes originating from single prothalli show that a genetic load is present in all population examined. It appears not to be a simple allelic load but a complicated balanced system. Morphological variability can be interpreted as the expression of the genetic heterogeneity of populations. There are no crossing barriers, not even between insular populations hundreds of kilometers apart.     

A quantitative comparison of foliage development among allopatric ferns,Dryopteris crassirhizoma,D. coreano-montana andD. filix-mas

A quantitative comparison was conducted on the foliage development during sporophyte development of three allopatric ferns in cool temperate and subalpine regions of Hokkaido and Tirol, European Alps. The foliage development ofDryopteris crassirhizoma, D. coreano-montana andD. filix-mas was quantitatively described by the leaf development (NV, number of veins); NV correlates the leaf-shape complexity from a circle (DI, L/2(3.14×S)^1/2). Nearly similar patterns were detected on frequency distribution of fertile leaves, fertility increase and number of leaves in threeDryopteris ferns which exhibit funnel-shaped foliage arrangements in mature sporophyte. No difference was found in number of leaves, maximum NV, fertility rate and leaf-shape parameters among three ferns. A positive difference was found only on changes in order of pinnae with maximum number of costa branches (NVP) and the DI of outline of pinnae betweenD. crassirhizoma andD. filix-mas. These allopatricDryopteris ferns seem to have a similar foliage structure, in spite of some sympatricDryopteris ferns capable of producing putative hybrids (D. austriaca andD. amurensis; D. tokyoensis andD. monticola) having different foliage structures in Hokkaido. Contribution No. 3346 from the Institute of Low Temperature Science, Hokkaido University.     

REM-Untersuchungen anMilesia vogesiaca undMilesia whitei (Uredinales)

Central European specimens of twoMilesia species, both growing onPolystichum, were investigated by SEM. The uredospores ofM. vogesiaca are not smooth, as stated in literature, but finely verrucose; size and density of the verrucae are rather variable. In contrast, the uredospores ofM. whitei are finely echinulate.

     

Intragametophytic selfing does not reduce reproduction inDryopteris filix-max

Gametophytes ofDryopteries filix-mas (L.) Schott were grown as isolates and as pairs of related and unrelated individuals. Reproductive efforts were generally highest among isolates, which reproduce by intragametophytic selfing only. Reproductive success ofD. filixmas may depend on the abundance of resources rather than the actual mating system employed. The estimates of the amount of genetic load for the two populations examined were low, 10% and 16%, respectively. The great numbers and the high viability of sporophytes produced by intragametophytic selfing indicate thatD. filix-mas does not experience serious inbreeding depression.     

Wood anatomy ofTrimeniaceae

Four collections of three species ofTrimenia and one collection ofPiptocalyx were studied; early-formed and later-formed wood was analyzed for oneTrimenia. Liquid-preserved material permitted analysis of mucilage and starch storage in wood ofT. neocaledonica andP. moorei. BecausePiptocalyx is scandent whereasTrimenia is arborescent, wood differences relative to evolution of a climbing habit could be examined.Piptocalyx contrasts withTrimenia in having wider vessels, more numerous per mm², resulting in a conductive area five times greater per unit area than that of theTrimenia woods averaged.Piptocalyx has appreciably fewer bars per perforation plate and thus much greater conductive area per perforation plate than have the species ofTrimenia. Rays inPiptocalyx are much taller and wider than those ofTrimenia. Wood ofTrimeniaceae is highly primitive in its scalariform perforation plates, scalariform lateral wall pitting on vessels, relatively long vessels elements, and heterocellular rays. Imperforate tracheary elements are septate nucleate fibertracheids (or even libriform fibers) rather than tracheids, but loss of borders on pits (and thus lowered conductive function of the imperforate tracheary elements) can be explained by the development of these elements into starchstoring cells. Some fiber-tracheids inT. neocaledonica are enlarged mucilagecontaining cells. Details of vessel structure inTrimeniaceae are similar to those ofMonimiaceae (s. s.), but similarity to some other lauralean (annonalean) families may be found: in mucilage presence,Trimeniaceae resembleLauraceae rather thanMonimiaceae. Wood ofTrimeniaceae may be regarded as highly mesomorphic, corresponding to the moist habitats in which all of the species occur.     

Allozymic and morphometric variation inLemna minor (Lemnaceae)

Allozymic and morphometric variation was studied in 28 clones ofLemna minor. This variation was compared with the corresponding variation in four clones ofLemna gibba and four clones ofSpirodela polyrrhiza. A high level of allozymic variation was observed among the clones, despite having been grown under uniform laboratory conditions for several years and despite its quasi-exclusive clonal means of propagation. Based on degree of allozymic similarity,Spirodela polyrrhiza was distinguished from the twoLemna species but the latter species were genetically indistinguishable. Allozymic similarity among clones ofLemna minor was not related to morphometric similarity, nor was it related to the degree of geographic separation or climatic similarity of their sites of origin. The results suggest that allozymic variation among these clones ofLemna minor may be largely neutral and not a consequence of differential selection.     

Biological species ofArmillaria and their mycoparasitic associations withRhodophyllus abortivus in Hokkaido

Specimens of basidiomes and/or rhizomorphs ofArmillaria mellea complex and basidiomes ofRhodophyllus abortivus, developing on the same decaying stumps or stems of forest trees, were collected in three forests in Hokkaido. Normal basidiomes ofR. abortivus were found near to, but free from, the rhizomorphs and/or basidiomes ofArmillaria, while abnormal basidiomes, as carpophoroid forms, were developed on the rhizomorphs ofArmillaria. Of three mycoparasiticArmillaria isolates found withR. abortivus, one was identified asA. gallica and two asA. jezoensis. The isolates ofR. abortivus showed excellent mycelial growth and rhizomorph formation on PDA. However, on MDA, RMDA and BMDA, they showed poor aerial mycelia growth and no rhizomorphs. In the contrapositional cultures, the growth ofA. gallica was completely inhibited byR. abortivus on PDA but only slightly inhibited on MDA and RMDA. On the other hand, mutual inhibition at a distance was observed on BMDA. The mycelial growth and rhizomorph formation inA. jezoensis were severely inhibited by the colony ofR. abortivus on PDA, but only slightly inhibited on MDA. On RMDA and BMDA, the colonies of twoArmillaria species andR. abortivus showed mutual inhibition at a distance and apparent rhizomorph formation by bothArmillaria species.     

Lead tolerance and accumulation in the gametophytes of the fern Athyrium yokoscense

The fern Athyrium yokoscense is known to be highly tolerant to lead toxicity, and is a lead hyperaccumulator that can accumulate over 1,000 g g^–1 of lead in its dry matter. In this work, we examined whether the gametophytic generation of A. yokoscense also resists lead toxicity like the sporophytic generation. Spore germination in A. yokoscense was more tolerant to Pb²⁺, compared to that in other fern species, such as Pteridium aquilinum, Lygodium japonicum and Pteris vittata. In addition, the early gametophyte development of A. yokoscense was not much affected by 10 M Pb²⁺, as evaluated from the prothallial growth and rhizoid development. We also showed that Athyrium gametophytes could accumulate more than 10,000 g g^–1 of lead, and that the lead was localized in the cytosol and vacuole of rhizoidal cells, as determined by a transmission electron micrograph. These results indicate that Athyrium gametophytes have the ability to accumulate lead in the rhizoids. Furthermore, the gametophytes were found to include a large amount of proanthocyanidins (condensed tannins). Because proanthocyanidins have a latent ability to complex with lead ions, the possible roles of proanthocyanidins in the lead tolerance and accumulation of Athyrium gametophytes are discussed.     

Zur Kenntnis vonTetrasporopsis fuscescens

The macroscopic colonies ofTetrasporopsis fuscescens consist of a tegument of irregularly folded bag-like mucilage. It contains isolated ellipsoidal cells arranged in a regular peripheral stratum and rather sparsely and irregularly beneath. They show polarity, 4 parietal chromatophors when fully developed, have a wall containing cellulose, and no contractile vacuoles. Naked protoplasts are released from them through circular openings oriented towards the surface. Flagellated cells and cysts were not observed.T. reticulata may be closely related toT. fuscescens, but probably no other species is allied.

Herrn Univ.-Prof. Dr.Walter Leinfellner zum 70. Geburtstag gewidmet.     

Mitochondrial DMA variation in somatic embryogenic cultures ofLarix

outhern hybridization analysis using wheat mitochondrial gene-specific probes indicates that changes in mitochondrial genomic organization and the relative representation of certain genomic regions occur during in vitro somatic embryogenic cell culture ofLarix species. We observed differences in the mitochondrial (mt)DNA hybridization patterns between somatic embryogenic cell cultures and trees grown from seed forLarix leptolepis,L. decidua, and the reciprocal hybrids of these twoLarix species. This is the first study to describe the correlation of molecular changes in a gymnosperm mitochondrial genome with in vitro somatic embryogenic cell culture. Quantitative differences in mtDNA hybridization signals were also observed among a 4-year-old somatic embryogenic cell culture ofLarix ×eurolepis trees regenerated from this culture, and the seed source tree from which the somatic embryogenic cell cultures were initiated.     

Competitive growth of slow growingRhizobium japonicum against fast growingEnterobacter andPseudomonas species at low concentrations of succinate and other substrates in dialysis culture

A cultivation system with simultaneous growth of six bacterial cultures in separate bags in dialysis culture was developed. In a medium with no added carbon source (one half concentrated Hoagland solution, water deionized and distilled), cell number ofRhizobium japonicum increased during a 7 day period by a factor of 35, whereas the number ofEnterobacter aerogenes cells decreased to one half. With a concentration of 100 nM succinate as an additional carbon source in the inflow,Rhizobium japonicum 61-A-101 cell number increased by a factor of 50 during an 8 day period, whereas cell number ofEnterobacter cloacae NCTC 10005 only doubled and ofEnterobacter aerogenes NCTC 10006 decreased. At 10 mM concentration of succinate in the inflow, doubling time the twoEnterobacter strains was about 12 h, compared to about 24 h for theRhizobium japonicum strain. Varying the succinate concentration from 10 mM to 100 nM in the inflow,Rhizobium japonicum 61-A-101 surpassed theEnterobacter aerogenes strains in the growth rate between 1 mM and 100 M succinate in the inflowing medium. Three otherRhizobium japonicum strains (fix⁺ and fix^-) did grow with a similar rate as strain 61-A-101 at very low concentrations of substrate. Growth rates for the strains were confirmed by protein data per culture. Growing in competition with twoPseudomonas strains,Rhizobium japonicum RH 31 Marburg (fix^-) did overgrow alsoPseudomonas fluorescens, was however outgrown byPseudomonas putida. In utilizing low concentrations of a¹⁴C labelled organic acid (malonate), three strains ofRhizobium japonicum left 2–4 times smaller amounts of¹⁴C in the medium than two species ofPseudomonas and two species ofArthrobacter.On sabbatical leave at ANU     

Localized hybridization betweenSaxifraga aizoides andS. mutata (Saxifragaceae): Reproductive ecology ofS. ×hausmannii and implications for conservation

The roles of hybridization and of introgression as potential threats to small native populations are current topics of interest both for evolutionary and conservation biologists. Two factors are mainly involved in the threat to natural populations through hybridization: demographic swamping and genetic assimilation. At prealpine locations, twoSaxifraga species with very different growth habits,S. aizoides andS. mutata, form a hybrid,S. ×hausmannii. Investigation of meiosis and crossing experiments revealed regular pairing of the 26 chromosomes and development of viable seeds in the hybrid. Analysis of hybrid indices, GPI allozymes, germination experiments and seed sizes showed an intermediate position ofS. ×hausmannii between its parental species.Saxifraga aizoides andS. mutata were separated by only weak hybridization barriers. Introgression occurred mostly in direction ofS. aizoides, although to a small amount. There is no actual threat to small, prealpine populations ofS. aizoides andS. mutata through interspecific gene flow because hybridization is localized, resulting from different flowering phenologies of the parental species and from the narrow habitat requirements ofS. ×hausmannii, which are similar to those ofS. aizoides. The major threat to local, prealpine populations ofS. aizoides andS. mutata is the loss of landscape dynamics. Both species need open erosion slopes for successful recruitment.     

Polar distribution of annexin-like proteins during phytochrome-mediated initiation and growth of rhizoids in the ferns Dryopteris and Anemia

Although the calcium requirement of phytochrome-mediated fern spore germination and early rhizoid growth is well established, the calcium-binding proteins that serve as transducers for these responses are not known. Here we report the presence of annexin-like proteins in germinating spores of Dryopteris filix-mas (L.) Schott and Anemia phyllitidis (L.) Sw. and evidence that they may be important participants in early photomorphogenic changes in gametophytes. Immunolocalization and immunoblot assays of these proteins were carried out using polyclonal antibodies raised either against a 35-kDa annexin-like protein from pea or against anchorin CII from chicken. Western-blot analysis showed that crude protein extracts obtained from both species after red-light treatment contained two cross-reactive protein bands with molecular weights around 70 kDa. These proteins were annexin-like in that they bound to a phosphatidylserine affinity column in a calcium-dependent fashion. Using this column, two protein bands around 70 kDa, i.e. 67 and 73 kDa, were partially purified together with proteins at 36 kDa and a doublet at 54 kDa. Proteins of these latter molecular weights are suggested to be members of the annexin family, but no cross-reactivity could be found between these and the two antibodies used in our investigations. Immunodetectable levels of these proteins were observed only after light-mediated induction of spore germination. Imaging of the immuno-localization patterns observed with both antibodies showed that the annexin-like proteins are concentrated at the extreme tips of the rhizoids in D. filix-mas and A. phyllitidis during rhizoid initiation and all stages of elongation. We suggest that these proteins may play a major role in the tip-oriented exocytosis events that are critical for the initiation and growth of fern rhizoids.Abbreviations CLSM confocal laser scanning microscopy - R red light Dedicated to Professor Andreas Sievers on the occasion of his retirementWe gratefully acknowledge the interest shown by Drs. D.-P. Häder (Botanisches Institut I, Erlangen, Germany) and M. Cresti (Dipartimento di Biologia Ambientale, Siena, Italy), and their generous provision of laboratory facilities. The authors thank Mrs. H. Klappstein and Mr. M. Becker for the careful preparation of the figures. This research was supported by a NATO collaboration grant (CRG 920082) to R. Scheuerlein and by a NASA grant (NAGW 1519) to S.J. Roux.