Abstract: The high mountain pottioid moss Molendoa hornschuchiana (Hook) Lindb. ex Limpr. is a very rare and critically endangered bryophyte species in Europe in need for ex situ conservation. A 25-year-old herbarium sample was used to revive and propagate this species for further reintroduction and introduction to potential natural habitats. The reviving of ‘‘dead’’ herbarium specimen was achieved by disposing of axenical organisms as well as adjusting condition for developing secondary protonema, bud inductions, and optimization of gametophyte propagation in vitro condition.The influence of exogenously added growth regulators on the morphogenesis of this species was studied. The plants were cultured in the two basic types of media, viz., BCD and half-strength Murashige and Skoog (MS) supplemented with different concentrations (0.01–0.3 mM) of indole-3-butyric acid (IBA) and N6-benzyladenine (BA) under a 16-h photoperiod. The influence
of growth regulators on gametophores multiplication in vitro as well as on protonemal diameter was recorded. Well-developed gametophores were obtained on BCD medium, whereas on half-strength MS medium, secondary protonema was produced, both on hormone-free and supplemented substrate exclusively. Based on multiplication index in vitro,maximumdevelopment of gametophoreswas realized on BCDmediumsupplemented with 0.3 mM IBA and 0.1 mM BA. However, the widest diameter of secondary protonema was obtained on BCD medium enriched with low concentration of both BA (0.01 and 0.03 mM) and constant concentration of IBA (0.03 mM). Chemical names used: indole-3-butyric acid (IBA), N6-benzyladenine (BA), Murashige and Skoog medium (MS).
Abstract: The study gives the first report of in vitro culture of the pleurocarpous moss Hypnum cupressiforme and on the problems of axenicaly culturing this bryophyte and the conditions for establishment and propagation. Problems of surface sterilization are elaborated regarding sporophyte vs. gametophyte. The influence of nutrient, light length and temperature on different developmental stages is discussed. The best conditions for micro-propagation from shoots are slightly lower temperature (18-20°C), on MS-sugar free medium irrelevant of day length. This moss is a counterpart of some rare and endangered mosses from the same genus and date presented should be taken into account of conservation and propagation of its counterparts as well. Its propagation is valuable for horticultural, pharmaceutical and bioindication purposes, as well.
Abstract: The use of in vitro techniques for conservation has been rising steadily since their inclusion in The Convention on Biological Diversity and The Global Strategy for Plant Conservation. Unfortunately, bryophytes are often overlooked in conservation initiatives, but they are
important in a number of large-scale ecosystem processes, i.e. nutrient, water and carbon cycling. There is a long history of the use of tissue culture in cultivating bryophytes, and many species respond well to in vitro techniques. For 6 yr (2000–2006), The Royal Botanic Gardens, Kew and the UK statutory conservation agencies supported a project for the ex situ conservation of bryophytes. Living and
cryopreserved collections of UK threatened species were successfully established and the cryopreserved collection continues to be maintained. Other in vitro conservation collections are maintained over Europe, at botanic gardens, museums and by individual university researchers, but there is no coherent European collection of bryophytes for conservation, or standardisation of techniques. A major issue for many in vitro collections is the maintenance of within species genetic diversity. Such diversity is considered to be important, as it is the basis by which populations of species can adapt to new conditions and evolve. We are proposing to establish a European network for in vitro conservation of bryophytes. We envisage that this will include living collections, cryopreserved collections and spore collections. Conservation of genetic diversity would be a priority and the collections would provide a valuable resource for conservation initiatives and support research into rare and threatened species.
Abstract: The antifungal activity of dimethyl sulfoxide (DMSO) extracts of three bryophyte species, two mosses (Atrichum undulatum (Hedw.) P. Beauv., Physcomitrella patens (Hedw.) Bruch and Schimp) and a liverwort (Marchantia polymorpha L. ssp. ruderalis Bischl. and Boisselier), grown in nature and in axenic culture was evaluated by microdilution method against five fungal species (Aspergillus versicolor, Aspergillus fumigatus, Penicillium funiculosum, Penicillium ochrochloron and Trichoderma viride). All the investigated bryophyte extracts have been proved to be active against all fungi tested. In general, extracts made from material grown in the laboratory (in vitro) conditions express better anti-fungal activity comparing to those made from material grown in the nature. Some of the fungi tested react similarly to both extracts.
Abstract: Background and Purpose: Cytokinins are a group of plant hormones
that have an important role in plant growth and developmental processes.
Chlorophyll content is an extremely important parameter in estimating the plant production level. Since bryophytes do not have such economical importance as vascular plants and their production in many ecosystems is small, they remain uninteresting for studying their chlorophyll level. The aim of this study was to compare the effect of different cytokinins on chlorophyll retention in moss B. argeteum gametophyte shoots grown in natural conditions with those grown in in vitro culture.
Material and Methods: The effect of different cytokinins: kinetin
(KIN), 6-benzylaminopurine (BAP) and thidiazuron (TDZ) on chlorophyll retention of the moss Bryum argenteum Hedw. (Bryaceae) derived from in vitro culture or grown in nature was tested. Plants derived from in vitro culture were grown on Murashige and Skoog (MS) medium at 25±2°C. Gametophyte shoots were used in experiments where influence of different concentrations (0.001–10 μM) of three cytokinins was used to investigate their effect on chlorophyll-a, -b and total chlorophyll retention.
Results and Conclusions: Cytokinins had a positive but unequal influence on chlorophyll retention in both plant groups – plants derived from in vitro culture and plants grown in the nature. Kinetin proved to be the most effective cytokinin in chlorophyll retention. Exogenous application of kinetin increased chlorophyll content with concentration (0–10μM). BAP had similar trends in in vitro and native mosses, increasing chlorophyll content up to 1 μM and then significantly decreasing, although the chlorophyll content was greater in in vitro grown plants. TDZ showed significantly better effect in in vitro cultured moss shoots, but when applied in concentrations
higher than 0.1 μM, total chlorophyll content decreased.
Abstract: The moss Bryum argenteum Hedw. was treated with gibberellins as well as some inhibitors of gibberellin
biosynthesis in order to investigate their influence on B. argenteum morphogenesis. Generally, gibberellins have not
been chemically identified in bryophytes, while other groups of classical phytohormones (auxins, cytokinins, abscisic
acid and ethylene) have been chemically identified in these plants. The in vitro culture of the moss Bryum argenteum
was established from sterilized spores. The apical shoots of untreated gametophytes grown in vitro were used to
investigate the influence of different substances on secondary protonema and on the growth and multiplication of the
gametophytes. B. argenteum reacts differently to the growth regulators applied. Both gibberellins applied in vitro (GA3
and GA7) have a positive effect on B. argenteum morphogenesis. Shoot multiplication was negatively affected by three
tested growth retardants (ancymidol, BX-112 and chlorocholine chloride), while these substances did not have such
strong effects on the moss protonema development.
Abstract: The antimicrobial activity of DMSO extracts of three bryophyte species, two mosses and a liverwort (Atrichum undulatum (Hedw.) P. Beauv., Marchantia polymorpha L. ssp. ruderalis Bischl. & Boisselier, Physcomitrella patens (Hedw.) Bruch & Schimp.) grown in nature and in axenic culture was evaluated by microdilution method against eight bacterial species (Escherichia coli ATCC 35210, Pseudomonas aeruginosa ATCC 27853, Salmonella typhimurium ATCC 13311, Enterobacter cloacae (human isolate), Listeria monocytogenes NCTC 7973, Bacillus cereus (human isolate), Micrococcus flavus ATCC 10240 and Staphilococcus aureus ATCC 6538).
All investigated bryophyte extracts are proved to be active against all bacteria tested. In general, extracts made from material grown in nature express better anti-bacterial activity comparing to those made from material grown in laboratory conditions. Some of the bacteria react the same to both extracts and some even better to the extracts made from axenically grown material.
Abstract: Moss species of the family Brachytheciaceae, Kindbergia praelonga (Hedw.) Ochyra and Kindbergia stokesii (Turn.) Ochyra, were preliminarily analysed for their fatty acid composition with the aim of studying the chemical relationship of these two entities. Fatty acid methyl esters were examined by GC and GC–MS in their methanol extracts. Thirteen fatty acids were identified.
It is likely that the mosses are chemically distinguishable and should be treated as separate entities. However, additional chemical constituents of various moss samples, such as phenolic acids, their derivatives and flavonoids, must be also analyzed in order to support the re-examination of the relationship between these two species.
Abstract: A moss genus Herzogiella, from the pleurocarpous family Plagiotheciaceae contains only seven species world wide. It occurs in North, Central and South America, Europe and Asia. In Europe, only three species occurred, namely H. seligeri, H. striatella and H. turfacea of which, the last one is threatened. With aim to develop the methodology for protection, conservation and active propagation of H. turfacea, more commonly distributed counterpart, H. seligeri, were taken from the National Park Fruška Gora and axenically culture were established. The study gives overview into the problems of sterilization, in vitro establishing, development, propagation and biology of species, as well as indices applicable to threatened counterpart.
Abstract: The moss Bryum argenteum Hedw. was treated with gibberellins as well as some inhibitors of gibberellin
biosynthesis in order to investigate their influence on B. argenteum morphogenesis. Generally, gibberellins have not
been chemically identified in bryophytes, while other groups of classical phytohormones (auxins, cytokinins, abscisic
acid and ethylene) have been chemically identified in these plants. The in vitro culture of the moss Bryum argenteum
was established from sterilized spores. The apical shoots of untreated gametophytes grown in vitro were used to
investigate the influence of different substances on secondary protonema and on the growth and multiplication of the
gametophytes. B. argenteum reacts differently to the growth regulators applied. Both gibberellins applied in vitro (GA3
and GA7) have a positive effect on B. argenteum morphogenesis. Shoot multiplication was negatively affected by three
tested growth retardants (ancymidol, BX-112 and chlorocholine chloride), while these substances did not have such
strong effects on the moss protonema development.
Abstract: In this study, the deposition of three heavy metals (Mn, Mo, and Ni) in the county of Obrenovac (Serbia) in four moss taxa (Bryum argenteum, Bryum capillare, Brachythecium sp., and Hypnum cupressiforme) is presented. The distribution of average heavy metal content in all mosses in the county of Obrenovac is presented on maps, while the long-term atmospheric deposition (in the mosses Bryum argenteum and B. capillare) and short term atmospheric deposition (in the mosses Brachythecium sp. and Hypnum cupressiforme) are discussed and given in tabular form. Areas of the highest contaminations are highlighted.
Abstract: In this study, the deposition of three heavy metals (Pb, Sr and Zn) in the county of Obrenovac (Serbia)
in four moss taxa (Bryum argenteum, Bryum capillare, Brachythecium sp. and Hypnum cupressiforme)
is presented. Distribution of average heavy metal content in all mosses in the county of Obrenovac is
presented in maps, while long term atmospheric deposition (in mosses Bryum argenteum and B.
capillare) and short term atmospheric deposition (in mosses Brachythecium sp. and H. cupressiforme)
is discussed and given in tables. Areas of the highest contaminations are highlighted.
Abstract: In this paper the effects of gypsum (CaSO4 x 2H2O) in the growth medium were tested on two
non-gypsophilous selected bryophyte species: Bryum argenteum Hedw. and Atrichum undulatum
(Hedw.) P. Beauv. With aim to test if some bryophytes are exclusive for gypsum enriched substrates,
the difference in gypsum effect on moss development were examined. In vitro cultures of two
selected species were initiated from nearly mature spores within unopened capsules. Half strenght
Murashige and Skoog media with added gypsum in concentrations of 50, 350 and 500mM were
used to test bryophyte develepment. Plants were grown on media with gypsum for 3 days or 3 weeks
to compare short and long term effect of salt exposure. Tested bryophytes, non-adapted to gypsum
and exposed to various gypsum concentrations in MS medium, did not show to have a problem
in surviving the new environment conditions. Moss species selected in this study and exposed to
gypsum enriched MS medium showed variation in morphological parameters to some extent (index
of multiplication, secondary protonemal production and survival rate) and chlorophyll content and
slightly altered chlorophyll a/b ratio. In general, both tested species could survive gypsum enriched
medium: secondary protonema was developed and new shoots were formed in both species, but
slight differences were recorded between short and long term exposure to gypsum. Acording to
data obtained, gypsum is not the stressor (at least not alone) which can separate some interesting
bryophytes into the ecological group of gypsophytes.
Abstract: In this paper the effects of gypsum (CaSO4 x 2H2O) in the growth medium were tested on two
non-gypsophilous selected bryophyte species: Bryum argenteum Hedw. and Atrichum undulatum
(Hedw.) P. Beauv. With aim to test if some bryophytes are exclusive for gypsum enriched substrates,
the difference in gypsum effect on moss development were examined. In vitro cultures of two
selected species were initiated from nearly mature spores within unopened capsules. Half strenght
Murashige and Skoog media with added gypsum in concentrations of 50, 350 and 500mM were
used to test bryophyte develepment. Plants were grown on media with gypsum for 3 days or 3 weeks
to compare short and long term effect of salt exposure. Tested bryophytes, non-adapted to gypsum
and exposed to various gypsum concentrations in MS medium, did not show to have a problem
in surviving the new environment conditions. Moss species selected in this study and exposed to
gypsum enriched MS medium showed variation in morphological parameters to some extent (index
of multiplication, secondary protonemal production and survival rate) and chlorophyll content and
slightly altered chlorophyll a/b ratio. In general, both tested species could survive gypsum enriched
medium: secondary protonema was developed and new shoots were formed in both species, but
slight differences were recorded between short and long term exposure to gypsum. Acording to
data obtained, gypsum is not the stressor (at least not alone) which can separate some interesting
bryophytes into the ecological group of gypsophytes.
Abstract: Photoperiod is known to regulate many essential processes in plants, but physiological effects of
photoperiod in vegetative stage of plant life have seldom been studied. This paper deals with effects
of day length on Catherine’s moss grown in aseptic culture. Photosynthetic pigments did not show
significant variations as a consequence of growth in different photoperiods. Protein content and
malate dehydrogenase activity were higher in long day (16h light/8h dark) than in short day (8h
light/16h dark) grown plants. Total phenolic compounds contents, as well as total antioxidative
capacity were shown to be higher in plants grown in long day conditions. Peroxidase activity was
also higher in long day than in short day grown plants. Regulation of components of antioxidative
metabolism in a moss species grown in different photoperiods are discussed in relation to same
parameters in higher plants.
Abstract: The first procedures have been established for in vitro micropropagation, plant regeneration, and protoplast transformation of the desiccation-tolerant resurrection plant, Ramonda serbica PanÄ. An integrated optimization of salt and growth regulator composition, and different culture temperatures resulted in an effective tissue culture system, which also prevented the undesirable hyperhydricity and tissue necrosis typical of Ramonda tissue cultures. De novo regenerated plants developed at high frequencies, which is a precondition for gene transfer and offer a good opportunity to optimize genetic transformation of R. serbica. Using leaf blades of in vitro propagated plants as donor tissue for protoplast isolation and GFP reporter gene as visually selectable marker gene, PEG-mediated protoplast transfection was also performed. This provides a new tool for studying cellular-level stress responses like stress associated membrane degradation, deficiencies in membrane transport processes (patch clamp technique), or cell cycling abnormalities.
Abstract: Recently, an alternative route to the proteasomal protein-degradation pathway was discovered that specifically targets transmembrane proteins marked with a single ubiquitin to the endosomal multivesicular body (MVB) and, subsequently, to the vacuole (yeast) or lysosome (animals), where they are degraded by proteases. Vps23p/TSG101 is a key component of the ESCRT I-III machinery in yeast and animals that recognizes mono-ubiquitylated proteins and sorts them into the MVB. Here, we report that the Arabidopsis ELCH (ELC) gene encodes a Vps23p/TSG101 homolog, and that homologs of all known ESCRT I-III components are present in the Arabidopsis genome. As with its animal and yeast counterparts, ELC binds ubiquitin and localizes to endosomes. Gel-filtration experiments indicate that ELC is a component of a high-molecular-weight complex. Yeast two-hybrid and immunoprecipitation assays showed that ELC interacts with Arabidopsis homologs of the ESCRT I complex. The elc mutant shows multiple nuclei in various cell types, indicating a role in cytokinesis. Double-mutant analysis with kaktus shows that increased ploidy levels do not influence the cytokinesis effect of elc mutants, suggesting that ELC is only important during the first endoreduplication cycle. Double mutants with tubulin folding cofactor a mutants show a synergistic phenotype, suggesting that ELC regulates cytokinesis through the microtubule cytoskeleton.
Abstract: The antimicrobial activity of Bryum argenteum ethanol extracts was evaluated by microdilution method against four bacterial (Escherichia coli, Bacillus subtilis, Micrococcus luteus and Staphilococcus aureus) and four fungal species (Aspergillus niger, Penicillium ochrochloron, Candida albicans and Trichophyton mentagrophyes). All the investigated ethanol extracts have been proved to be active against all bacteria and fungi tested.
Abstract: The effect of different sugars (sucrose, fructose and glucose) on morphogenesis of two moss species, Bryum argenteum Hedw. (Bryaceae) and Atrichum undulatum (Hedw.) P. Beauv. (Polytrichaceae) in in vitro culture was tested. In vitro cultures of these two moss species were established from disinfected spores on Murashige and Skoog (MS) medium. Apical shoots of well-developed gametophytes were used to investigate the influence of different concentrations (0.01 M, 0.03 M, 0.05 M, 0.1 M, 0.2 M, 0.3 M) of sugars on secondary protonemal and gametophyte growth and multiplication. In in vitro culture of B. argenteum sugars had a positive influence on protonemal development and shoot multiplication, while in in vitro culture of A. undulatum these two processes were negatively correlated with addition of all tested sugars. Although all exogenous sugars tested affected developmental processes in B. argenteum and A. undulatum in in vitro conditions, these two moss species reacted differently.
Abstract: Bryophytes are the second biggest group of terrestrial plants. They have not
received much attention in biotechnology due to their small size and biomass.
However, lately many new features have been highlighted and the interest
for the potentials use of these plants increased. Scientifically new chemical
compunds are described from bryophytes. Bryophyte extracts are shown to
be biologically active with particular reference for their use in pharmacology,
medicine and agriculture for all round benefit of living beings. There are
some reports that some bryophyte extract are able to cure or prevent deseases
such as AIDS and cancer. With axenic culturing of some species, the problems
of enough mass and clean bryophyte culture are partly solved. Due to their
properties and life span where the dominant life phase is haploid, bryophytes
represent easy tools for molecular and genetic investigation. The model
bryophyte plant whose genome has been sequenced is Physcomitrella patens.
Bryophytes also have extraordinary values in environmental bioindication,
or microdust attachment. They received attention as ornamental plants and
covering plants in restoration and landscape ecology. Still, many things
remain to be proven in the biology of these organisms. This chapter gives
the overview of the biotechnological use of bryophytes today and their further
use potentials.
Abstract: Bryophytes, the second largest group of land plants, are extremely rich in terpenoids, phenols, glycosides,
and fatty acids. Although bryophytes could be used in medicine, their chemistry is not very well known
and the problem remains to obtain enough quantity of same species for analysis. In vitro cultivation of
bryophytes is the most appropriate way for large biomass production and isolate of numerous useful
compounds showing some interesting biologic activities. This technique is also useful in developmental,
cellular, molecular, biochemical, and eco-physiologic studies.
