Pervasive RNA Editing Among Hornwort rbcL Transcripts Except Leiosporoceros

RNA editing affecting chloroplast and mitochondrial genomes has been identified in all major clades of land plants. The frequency of edited sites varies greatly between lineages but hornworts represent an extreme in propensity for editing in both their chloroplast and mitochondrial genomes. cDNA sequences from seven taxonomically diverse hornwort rbcL sequences combined with a survey of 13 additional DNA sequences for potential edited sites demonstrate the presence of 62 edited sites and predict a minimum of 10 additional sites. These 72 total edited sites represent 43 C-to-U and 28 U-to-C nucleotide conversions, with 1 site exhibiting editing in both directions. With one exception, all taxa are heavily edited, with each having from 20 to 34 edited sites. However, a single sample, Leiosporoceros, is shown to lack edited sites. Phylogenetic reconstruction of hornworts results in ambiguous resolution of Leiosporoceros depending on whether edited sites are maintained or eliminated from the analyses. Depending on the inferred relationship of Leiosporoceros to the hornworts, at least two explanations for the origin and maintenance of pervasive editing in hornworts are possible. The absence of edited sites in Leiosporoceros could represent either the absence or a low level of editing ability in the common ancestor of hornworts, as represented by Leiosporoceros, or the loss of editing sites in this lineage after the primary diversification events in the group.     

Sur la culture des spores et le développement du thalle de Botryocladia chiajeana (Menegh.) Kylin (Rhodyméniales,Rhodyméniacées)

Abstract

On the culture of spores and the development of the thallus of Botryocladia chiajeana (Menegh.) Kylin (Rhodymeniales, Rhodymeniaceae). - The complete development of Botryocladia chiajeana has been studied in culture. Spore germination and sporeling development is of the discalis mediatus type described by Inoh (1947). The hollow vesiculous branches filled with a mucilage derive from the spacing of the erect filaments which form the stipe. The authors compare this development of the thallus with that described for other Rhodymeniales.     

THE ECOLOGY OF NOSTOC

Nostoc is a genus of filamentous cyanobacteria that can form macroscopic or microscopic colonies and is common in both terrestrial and aquatic habitats. Much of the success of Nostoc in terrestrial habitats is related to its ability to remain desiccated for months or years and fully recover metabolic activity within hours to days after re-hydration with liquid water . Nostoc can also withstand repeated cycles of freezing and thawing and, thus, is an important component of extreme terrestrial habitats in the Arctic and Antarctic. The ability to fix atmospheric N ₂ can provide an advantage in nitrogen-poor environments . Nostoc also has the ability to screen damaging ultraviolet light in terrestrial and shallow benthic habitats. The genus potentially could be important in paddy rice culture because it fixes nitrogen that may later be released and used by plants; it also may play a role in soil formation and may increase nitrogen input to natural aquatic and terrestrial ecosystems. The abilities to survive in terrestrial habitats and fix N ₂ are important in symbiotic interactions with fungi (lichens), liverworts, hornworts, mosses, ferns, cycads, and the angiosperm Gunnera. Nostoc is somewhat resistant to predation; this probably is related to production of large amounts of sheath material, synthesis of microcystin-like toxins by some strains, and formation of colonies that are too large for many algivores to consume. Some organisms can subsist on Nostoc, although it may not be a preferred food source. Lytic cyanophages also infect Nostoc, but little is known about population control of Nostoc in its natural environment, Late Precambrian fossils resembling Nostoc have been described, and Nostoc possibly has been an important component of many terrestrial and aquatic communities since that time .     

On the mucilage canal and the mucilage gland in two species ofLaminaria (Phaeophyceae,Laminariales)

Phenological observations of mucilage canals inLaminaria angustata in field collected materials indicate that the initials of canals appear first in the spring in the transition zones in fronds of about 20 cm or more. They first appear as scattered dots. These initials are pshed upwards, elongating and branching and a broken mesh system of canals is finally formed as a result of stipe-frondal growth. Later, initials appear abundantly and continuously in the same zone, gradually producing a completely connected anastomosing system of canals. InLaminaria yezoensis, in one-cell-layered sporophytes, some of the component cells become initials of mucilage glands (fucosan cells), as seen inAlaria andUndaria. This phenomenon is confined to a very short-lived stage. Subsequently, mesh-like mucilage canals are formed as in most other Laminariaceous species as the plants grow. Anatomically, formation of mucilage canals in the two species takes place through secretory cells produced from periclinal division of the epidermal cells, followed by collapse and reorganization of groups of cells into mucilage canals. L. yezoensis, having both mucilage canals and mucilage glands in its development, may be classed at the border line betweenUndaria andAlaria which possess only the mucilage gland and other species ofLaminaria which possess the mucilage canal only.     

Artificial sporeling and field cultivation of Gelidium in China

The supply of Gelidium resources is dependent on collection from natural habitats. As these resources are very limited, one possible means of augmenting production is by implementation of artificial cultivation as has been done with Laminaria, Porphyra and Eucheuma. Chinese phycologists have conducted research on sporeling and field cultivation of Gelidium for many years. Trials have been made using: (1) raft cultivation based on vegetative propogation of thallus fragments, (2) cultivation based on spore-collecting and land-based tank culture to provide seedstock, (3) cultivation based on regeneration from small thallus fragments to provide seedstock. Some of these results have been promising, but development remains at the experimental stage and methods of cultivation need to be improved. This paper is an updated review of the research and development on artificial sporeling production and field cultivation of Gelidium in China.     

Microscopic observations on tissue calcium distribution in the brown alga,Undaria pinnatifida (Harvey) Suringar

The distribution of Ca²⁺ in thallus tissues of the brown algaUndaria pinnatifida was investigated by microspectro-photometric analysis after fixation with Carnoy's reagent and staining with an alizarine sulfonate dye. Ca²⁺ accumulation in the young thallus was much higher in mucilage cells of the basal than the apical region; there was least in the pith of the basal region. A roughly uniform pattern was found in the stipe from the central part of the young thallus, while a highly irregular pattern occurred in both stipe and blade from the corresponding area of the mature thallus; the Ca²⁺ content was much lower than in the cortex, mucilage cells and vacuoles of the mature stipe than in the corresponding area of the young stipe. The contents of all regions of the mature blade were higher than those in the young blade; they were particularly high in mucilage cells.     

Thallus initiation and development in the lichen Rhizocarpon lecanorinum

Ascospore germination, thallus initiation, and areole and prothallus development in the lichen Rhizocarpon lecanorinum Anders were examined using light, fluorescence and scanning electron microscopy. The ascospore germ hyphae remain very short and do not form a prothallus-like mycelium. Instead, a compact soredium-like granule develops directly from sporeling contact with a compatible species of Trebouxia . Diffuse initial stages involving non-trebouxioid algae are lacking. The onset of thallus differentiation is marked by the deposition of rhizocarpic acid in an incipient cortical layer within the apical part of the granule. As pigmentation and cortex-formation transform this structure into a typical areole, radiating prothallus hyphae are simultaneously initiated from its basal margin. Most areoles formed subsequently in the marginal prothallus lack subtending melanized hyphae and apparently stem from overgrowth by the prothallus of photobiont cells on, or in, the substratum. Apothecia reach maturity in thalli as small as 2 mm in diameter. It is proposed that the lack of diffuse hyphal growth in sporelings and telescoped morphogenesis of R. lecanorinum are part of a life history strategy geared to precocious, heavy investment in ascospore production. The R. lecanorinum – Trebouxia symbiosis has a number of features which make it well-suited for further studies of the life history and development of prothallus-forming crustose lichens with sexually reproducing mycobionts.     

A highly differentiated glomeromycotean association with the mucilage-secreting, primitive antipodean liverwort Treubia (Treubiaceae): clues to the origins of mycorrhizas

Thallus anatomy in three species of the primitive liverwort genus Treubia (Metzgeriidae, Treubiales) was studied by light and electron microscopy. The thallus exudes copious mucilage, a feature shared elsewhere in liverworts only with the mycotrophic subterranean axes of the allied genus Haplomitrium. The central strand in the thallus midrib has a unique histological organization and harbors an intra- and intercellular infection by a glomeromycotean fungus that is far more highly differentiated than most of the glomeromycotean associations described to date. The fungus enters the thallus via clefts in the ventral epidermis along the midrib and colonizes the parenchyma above, forming intracellular coils and prominent, relatively short-lived, hyphal swellings. Above the zone with intracellular colonization is a tissue area containing mucilage-filled intercellular spaces; here the fungus is entirely intercellular and forms abundant pseudoparenchymatous structures and, in more mature parts of the thalli, large hyphae with thick multistratose walls. Mucilage in Treubia differs in histochemistry and origin from that produced by apical papillae, via hypertrophied Golgi, in all other bryophytes. Remarkable parallels between fungal associations in Treubia, Haplomitrium, and Lycopodium, all members of very ancient lineages, suggest that these associations epitomize very early stages in the evolution of glomeromycotean symbioses.     

Inheritance of mucilage canals in Laminaria (section Simplices) in eastern Canada

Two experiments have been carried out to test the suitability of the occurrence of mucilage canals as a criterion for species or ecotype definition in the non-digitate section of the genus Laminaria. A matrix of crossability tests shows complete interfertility between all mucilage canal types, fertile F₁ hybrids being produced in all cases. Quantitative genetic analysis reveals a large environmental component in the phenotypic variance of degree of mucilage canal development. Only plants from Nova Scotia bred true with respect to mucilage canals. This characteristic is therefore considered generally unsuitable for taxonomic and biological species determination, though there may be evidence for intraspecific genotypic differentiation of Nova Scotian populations.     

A CYANOPHYTE CAPABLE OF FIXING NITROGEN UNDER HIGH LEVELS OF OXYGEN1

Nostoc cordubensis Prosperi (Cyanophyta, Nostocaceae) is characterized by its mucilaginous colonies. Much of this mucilage is produced by heterocysts. By controlling growth conditions, heterocysts with and without mucilage were obtained. Mucilaginous heterocysts retained nitrogen-fixing capability at high oxygen concentrations, whereas heterocysts lacking mucilage were unable to fix nitrogen at oxygen concentrations higher than 20%. Comparison of results obtained using tetrazolium salts as indicators of highly reduced zones showed similar results.     

The Structure and Development of the Thallus in the British Species of Gelidium and Pterocladia

The apical structure and the development of the thalli of allthe British species of Gelidium and Pterocladia have been investigated;the development of G. pulchellum is described in detail. Eachaxis is terminated by one or more apical cells, which by theirsegmentation form the tissues of the thallus. An axial filamentis distinct for a short distance behind each apical cell, butsecondary pit-connexions develop rapidly so that in sectionthe mature axis has the appearance of a multi-axial structure. Lateral branches of the frond develop by the segmentation ofthe lateral branch apical cells, which are formed by the transformationof superficial cortical cells, either in the meristematic ormature parts of the axes. The extreme variability of externalappearance is due principally to the indeterminate origin ofall lateral branches. The thallus in the British species of Gelidium and Pterocladiaconsists of erect fronds borne on creeping axes. The relativeproportions of the frond and creeping axes in various speciesand their survival through adverse conditions are discussed.     

Fine Structure and Biomineralization of the Mucilage in Envelopes of Trachelomonas lefevrei (Euglenophyceae)1

Envelope development in Trachelomonas lefevrei (Deflandre) begins with the production of short, coarse mucilaginous strands, morphologically similar to the mucilage produced by non-enveloped euglenoids. These initial mucilaginous secretions subsequently become impregnated with manganese (Mn) and/or iron (Fe). Continued mucilage secretion and mineralization results in a mature envelope that is characteristic for the species. When these mature envelopes are treated with oxalic acid to remove the Mn and Fe, the envelopes collapse and are composed only of short, coarse mucilaginous strands similar to those present during early stages of development, prior to their mineralization. Brief treatment with 10 mM EDTA renders dark envelopes colorless, and our EM-EDS analyses show that this corresponds to a loss of Mn from the envelope; however, if Fe is present in the envelope, it is unaffected by the brief treatment. The mucilage present during early stages of envelope development and that remaining after complete demineralization is also morphologically similar to the mucilage in the plug at the anterior end of the envelope.     

Extremotolerance and Resistance of Lichens: Comparative Studies on Five Species Used in Astrobiological Research I. Morphological and Anatomical Characteristics

Lichens are symbioses of two organisms, a fungal mycobiont and a photoautotrophic photobiont. In nature, many lichens tolerate extreme environmental conditions and thus became valuable models in astrobiological research to fathom biological resistance towards non-terrestrial conditions; including space exposure, hypervelocity impact simulations as well as space and Martian parameter simulations. All studies demonstrated the high resistance towards non-terrestrial abiotic factors of selected extremotolerant lichens. Besides other adaptations, this study focuses on the morphological and anatomical traits by comparing five lichen species—Circinaria gyrosa, Rhizocarpon geographicum, Xanthoria elegans, Buellia frigida, Pleopsidium chlorophanum—used in present-day astrobiological research. Detailed investigation of thallus organization by microscopy methods allows to study the effect of morphology on lichen resistance and forms a basis for interpreting data of recent and future experiments. All investigated lichens reveal a common heteromerous thallus structure but diverging sets of morphological-anatomical traits, as intra-/extra-thalline mucilage matrices, cortices, algal arrangements, and hyphal strands. In B. frigida, R. geographicum, and X. elegans the combination of pigmented cortex, algal arrangement, and mucilage seems to enhance resistance, while subcortex and algal clustering seem to be crucial in C. gyrosa, as well as pigmented cortices and basal thallus protrusions in P. chlorophanum. Thus, generalizations on morphologically conferred resistance have to be avoided. Such differences might reflect the diverging evolutionary histories and are advantageous by adapting lichens to prevalent abiotic stressors. The peculiar lichen morphology demonstrates its remarkable stake in resisting extreme terrestrial conditions and may explain the high resistance of lichens found in astrobiological research.     

The origin of the sporophyte shoot in land plants: a bryological perspective

Background

Land plants (embryophytes) are monophyletic and encompass four major clades: liverworts, mosses, hornworts and polysporangiophytes. The liverworts are resolved as the earliest divergent lineage and the mosses as sister to a crown clade formed by the hornworts and polysporangiophytes (lycophytes, monilophytes and seed plants). Alternative topologies resolving the hornworts as sister to mosses plus polysporangiophytes are less well supported. Sporophyte development in liverworts depends only on embryonic formative cell divisions. A transient basal meristem contributes part of the sporophyte in mosses. The sporophyte body in hornworts and polysporangiophytes develops predominantly by post-embryonic meristematic activity.

Scope

This paper explores the origin of the sporophyte shoot in terms of changes in embryo organization. Pressure towards amplification of the sporangium-associated photosynthetic apparatus was a major driver of sporophyte evolution. Starting from a putative ancestral condition in which a transient basal meristem produced a sporangium-supporting seta, we postulate that in the hornwort–polysporangiophyte lineage the basal meristem acquired indeterminate meristematic activity and ectopically expressed the sporangium morphogenetic programme. The resulting sporophyte body plan remained substantially unaltered in hornworts, whereas in polysporangiophytes the persistent meristem shifted from a mid-embryo to a superficial position and was converted into an ancestral shoot apical meristem with the evolution of sequential vegetative and reproductive growth.

Conclusions

The sporophyte shoot is interpreted as a sterilized sporangial axis interpolated between the embryo and the fertile sporangium. With reference to the putatively ancestral condition found in mosses, the sporophyte body plans in hornworts and polysporangiophytes are viewed as the product of opposite heterochronic events, i.e. an anticipation and a delay, respectively, in the development of the sporangium. In either case the result was a pedomorphic sporophyte permanently retaining juvenile characters.     

SPORELING COALESCENCE AND INTRACLONAL VARIATION IN GRACILARIA CHILENSIS (GRACILARIALES,RHODOPHYTA)1

This study evaluates the hypothesis that spore coalescence may cause intraclonal variation. Spore coalescence might allow the occurrence of unitary thalli that in fact correspond to genetically different, coalesced individuals. Plant portions simultaneously derived from these chimeric individuals may exhibit dissimilar growth responses even when incubated under similar abiotic conditions. Testing of the hypothesis included various approaches. Transmission electron microscopy observations of early stages of sporeling coalescence indicated that polysporic plantlets were formed by groups of spores and their derivatives. Even though adjacent cells in two different groups may fuse, these groups maintained an independent capacity to grow and form uprights. Laboratory-grown plantlets showed a significant correlation between the initial number of spores and the total number of erect axes differentiated from the sporeling. Construction and growth of bicolor individuals indicated the chimeric nature of the coalesced individuals. Coalesced, bicolor holdfasts had green and red cells, which subsequently produced green and red uprights, respectively. Individuals fronds were also chimeric, as indicated by the production of green and red branchlets from single, red uprights. The existence of mixed tissues was further substantiated by random amplified polymorphic DNA analysis. The banding pattern produced by branchlets of a unisporic thallus was consistently monomorphic, whereas the patterns produced by the polysporic thallus were polymorphic. Growth rates of polysporic thalli had larger data dispersal and variation coefficients than oligosporic or monosporic thalli. Therefore, all results support the original hypothesis and suggest that coalescence might be ecologically more important than previously thought.     

The Life-History of Marginariella urvilliana (Ach. Rich.) Tandy

In Marginariella urvilliana gametes are produced in the thirdyear and then annually for several years. On germination ofthe egg a single primary rhizoid is formed. Its early subdivisioninto four main branches is unusual amongst the Fucaceae. Growthof both the main axis and of the receptacles is carried outby a three-sided apical cell. The oogonia develop earlier thanthe antheridia. Nuclear divisions are completed within the conceptacle,often several weeks before extrusion of oospheres. The liberatedoosphere is held in a tubular stalk until after fertilizationand the sporeling has begun to develop. Marginariella has affinities with the other endemic submergedAustralasian fucoids, and together these form a rather isolatedgroup, probably rightly included within the Fucaceae.     

Phenotypic and Genotypic Comparison of Symbiotic and Free-Living Cyanobacteria from a Single Field Site

PCR amplification techniques were used to compare cyanobacterial symbionts from a cyanobacterium-bryophyte symbiosis and free-living cyanobacteria from the same field site. Thirty-one symbiotic cyanobacteria were isolated from the hornwort Phaeoceros sp. at several closely spaced locations, and 40 free-living cyanobacteria were isolated from the immediate vicinity of the same plants. One of the symbiotic isolates was a species of Calothrix, a genus not previously known to form bryophyte symbioses, and the remainder were Nostoc spp. Of the free-living strains, two were Calothrix spp., three were Chlorogloeopsis spp. and the rest were Nostoc spp. All of the symbiotic and all but one of the free-living strains were able to reconstitute the symbiosis with axenic cultures of both Phaeoceros and the liverwort Blasia sp. Axenic cyanobacterial strains were compared by DNA amplification using PCR with either short arbitrary primers or primers specific for the regions flanking the 16S-23S rRNA internal transcribed spacer. With one exception, the two techniques produced complementary results and confirmed for the first time that a diversity of symbiotic cyanobacteria infect Phaeoceros in the field. Symbionts from adjacent colonies were different as often as they were the same, showing that the same thallus could be infected with many different cyanobacterial strains. Strains found to be identical by the techniques employed here were often found as symbionts in different thalli at the same locale but were never found free-living. Only one of the free-living strains, and none of the symbiotic strains, was found at more than one sample site, implying a highly localized distribution of strains.     

ERUMPENT CEPHALODIA,AN APPARENT CASE OF PHYCOBIAL INFLUENCE ON LICHEN MORPHOLOGY1,2,3

On the upper surface of some specimens of the lichen Lobaria cf. crosa (Eschw.) Nyl. there are shrub-like growths which have a morphology similar to that of a free-living fruticose lichen, Polychidium umhausense (Auersw.) Henss., and contain a cyanophyte phycobiont, Nostoc. A few growths also contain scattered colonies of a chlorophyte phycobiont, in which case the lichen tissue locally assumes the foliose form characteristic of the parent Lobaria thallus. The differentiation of dorsiventral lichen cortices and the formation of a lax medulla and distinct algal layer are correlated with the presence of the green phycobiont. The lichen substances atranorin, gyrophoric acid, and 4-0-methylgyrophoric acid occur in both, the foliose L. erosa thallus and the fruticose tissue. It is suggested that the fruticose structures are erumpent cephalodia which are derived from the outgrowth of internal, cephalodia and should not be considered, to be epiphytic colonies of the lichen Polychidium.