Gibberellin influence on the morphogenesis of the moss Bryum argenteum Hedw. in in vitro conditions

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.Projekat ministarstva br. 143031 i 14301

Back to Hungary: a story on reintroduction of a European Habitat Directive moss species, Hamatocaulis vernicosus

Hamatocaulis vernicosus is rare and threatened species Europe-wide, and it is a member of many national red lists. The main reason for this can be regarded as habitat quality changes. The last records from Hungary comes from 1968, and in years after despite of intensive search it was not found in any historically known habitats. Thus, it was considered as extinct in Hungary. An attempt on getting back this species to Hungary has been made. Less than one year old herbarium specimens from neighboring countries have been used to establish new population but with limited success. The same material that could be revived has been used to start axenix in vitro culture. Once the culture has been established, the optimization and propagation started. The lab originated material was grown under controlled condition but in xenic medium originated from Hungary. Finally, after two years the moss plants were reintroduced to two national parks in Hungary and two populations were established. This is just the first report but there are continuous attempts to stabilize the populations. The problems, achievements and solutions will be discussed. A Hamatocaulis vernicosus, egy ritka és veszélyeztetett faj Európában és számos nemzeti vörös listán is szerepel. A fő veszélyeztető tényező az élőhelyeinek, a lápréteknek, a visszaszorulása, illetve leromlása. Az utolsó magyarországi record 1968-ból származik és bár az utóbbi években a közösségi jelentőségű fajok monitorozása intenzíven folyik, a Hamatocaulis vernicosus nem került elő egyetlen korábban ismert élőhelyéről sem. Így Magyarországról kipusztultnak tekinthető. Ennek a fajnak a visszatelepítésére tettünk kísérletet. Szomszédos országokból származó, egy éven belül gyűjtött herbáriumi példányokat próbáltunk meg felhasználni új populáció létrehozására, de ez csak korlátozott eredménnyel járt. Az egyik ilyen herbáriumi anyagból azonban sikerült in vitro kultúrát létrehozni és felszaporítani. A laborban kontrollált körülmények között tartott kultúrát magyarországi lápokról származó médiumon neveltük. Végül, két év után visszatelepítést végeztünk két nemzeti parkban és így két lápon sikerült egy-egy populációt létrehozni. A populációk stabilizálásán folyamatosan dolgozunk, nyomonkövetjük a változásokat. Bemutatásra kerülnek az elért eredmények, a felmerült problémák és lehetséges megoldások

In vitro culture and apogamy: Alternative pathway in the life cycle of the moss Amblystegium serpens (Amblystegiaceae)

In vitro culture of the moss Amblystegium serpens (Amblystegiaceae) was established on hormone-free Murashige and Skoog (MS) medium that contained a half amount of MS micro- and macro- mineral salts and vitamins, 100 mg/l myoinositol, 30 g/l sucrose, and 0.70% (w/v) agar. Spores were germinated and primary protonema developed on the above medium at 16 h day/8 h night 25±2ºC, 60-70% air humidity, and irradiance of 47 μmol/m2s. Three months after development of primary protonema, seven sporophytes appeared directly from primary protonema without generation alternation. The phenomenon of apogamous sporophyte formation is very rare, both in nature and under in vitro conditions. This is the first report of apogamy induced by Amblystegium serpens

Development of the moss Pogonatum urnigerum (Hedw.) P. Beauv. under in vitro culture conditions

Pogonatum urnigerum (Polytrichaceae) in vitro culture was established from spores collected in nature. Both protonema and gametophore stages of gametophyte development were obtained. Also, a stable callus culture was established using hormone-free nutrient medium. The best nutrient medium for development was half-strength Murashige-Skoog medium supplemented with 1.5% sucrose. Auxin treatment enabled some gametophores to develop, but prolonged treatment induced early senescence. Tissues grown on cytokinin did not produce any gametophytes and did not survive prolonged treatment.Sterilna kultura mahovine Pogonatum urnigerum (Polytrichaceae) uspostavljena je iz spora sakupljenih u prirodi. Gajenjem na hranljivoj podlozi bez biljnih hormona iz spora su se razvile protoneme i gametofiti. Uspostavljena je i stabilna kultura kalusa. Ustanovljeno je da je za gajenje ove vrste najbolja hranljiva podloga koja sadrži smanjenu koncentraciju soli i vitamina po Murashige-Skoog-u i 1.5% saharozu. Tretman auksinom nije pospešio razviće gametofita, dok je produženi tretman doveo do postepenog odumiranja protoneme. Tkiva gajena na podlozi koja je sadržala citokinine uopšte nisu produkovala gametofite i nisu preživela produženi tretman ovim hormonom. Ovaj rad predstavlja prvu studiju razvića vrste Pogonatum urnigerum u in vitro kulturi.nul

Effects of selected bryophyte species extracts on microorganisms

Two dozen of bryophyte species was collected from native habitat types across Europe to test if the ethanol extract can affect the growth of various microorganisms. After start-up test the ethanol extract did not show any influence on growth and development to Gram-negative bacteria and fungi Candida albicans (ATCC10231). Thus, the further tests focused on the Gram-positive bactreia. The ethanol extract of 5g bryophyte material was macerated in liquid nitrogen, and then extract was evaporated till dry. The dry residue was dissolved in 5ml of methanol. This was used in further analyses by MIC and MBC techniques. The result obtained showed the most effective extracts were those made from Pellia endiviifolia and Bazzania trilobata. Phytopathogenic bacteria and fungi C. albicans showed resistance to extracts of any bryophyte species tested. Gram-positive bacterias, namely Staphylococcus aureus (ATCC25923), Listeria monocytogenes (ATCC19111) and Bacilus subtilis (ATCC6633) were intolerant to extracts of liverworts P. endiviifolia and B. trilobata. MIC value for two above mentioned liverworts extract tested were 7-12mm, while referent antibiotic (rifampicin) inhibitory zone were 15-35mm. The most resistant to any bryophyte extracts was Escherichia coli (ATCC25922), while the most sensitive was B. subtilis (ATCC6633) with MIC values obtained 0.01-0.19 mg/ml. The results obtained show clear antimicrobial potential of P. endiviifolia and B. trilobata, however further research are needed

Effects of different cytokinins on chlorophyll retention in the moss Bryum argenteum (Bryaceae)

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

Osvrt na hemiju masnih kiselina Rhytididelphus squarrosus (Hedw.) Warnst

The fatty acid composition of the moss Rhytididelphus squarrosus (Hedw.) Warnst. (Hylocomiaceae) collected in Germany during winter time was analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Nine fatty acids were identifi ed in its chloroform:methanol extract 1:1: arachidonic acid (30.7%), α-linolenic acid (19.1%), linoleic acid (15.1%), cis-5,8,11,14,17-eicosapentaenoic acid (14.4%), palmitic acid (11.9%), cis-8,11,14- eicosatrienoic acid (4.1%), oleic acid (2.3%), γ-linolenic acid (1.4%) and stearic acid (1.0%). Th e results indicate that this plant species can be a good source of arachidonic acid collected during the winter.Sastav viših masnih kiselina mahovine Rhytididelphus squarrosus (Hedw.) Warnst. (Hylocomiaceae), sakupljene u Nemačkoj za vreme zimskog doba, preliminarno je ispitivan GC i GC-MS analizom. U njenom ekstraktu hloroform: metanol 1:1 identifikovano je devet viših masnih kiselina: arahidonska kiselina (30.7%), α-linolenska kiselina (19.1%), linolna kiselina (15.1%), cis-5,8,11,14,17-eikosapentaenska kiselina (14.4%), palmitinska kiselina (11.9%), cis-8,11,14-eikosatrienska kiselina (4.1%), oleinska kiselina (2.3%), γ-linolna kiselina (1.4%) i stearinska kiselina (1.0%). Dobijeni rezultati ukazuju da je ova biljna vrsta dobar izvor arahidonske kiseline u ispitivanom vremenskom periodu