Neuritogenic Activities of Various Kappaphycus alvarezii Extracts in Hippocampal Neurons

The carrageenophyte Kappaphycus alvarezii seaweed is known to have neuritogenic activities. Post-harvest treatment of seaweed affects its biological activities. Five drying treatments of K. alvarezii, including oven-drying, sun-drying, freeze-drying, shade-drying, and salting followed by shade-drying, were evaluated for their effects on neuritogenic activities. We also evaluated the neuritogenic activity of different parts of K. alvarezii thalli and its carrageenan waste. Neuron cells of the 19th day pregnant rat fetuses were collected from the hippocampus by brain dissection. Neuron cells were isolated by dissociation of the hippocampal tissue. Cells were plated onto poly-DL-lysine-coated glass coverslips in 24-well plates and treated with extracts. All tested extracts were obtained from maceration using 95% ethanol. Freeze and shade-drying extracts exhibited significantly higher neuritogenic activities (p 0.05) compared to that of the vehicle control. Carrageenan waste also significantly promoted the neuritogenic activities (p 0.05) with an optimal dose at 1 µg mL-1. Old and young thalli showed insignificant differences in neuritogenic activities. The carrageenan waste extract retained neuritogenic activities. Thus, the utilization of carrageenan waste for neuritogenic material provides added value to the waste in the carrageenan industry. Freeze and shade dried K. alvarezii can be used as a neuritogenic agent to provide optimum biological activity.

First Restoration Experiment for Gongolaria barbata in Slovenian Coastal Waters. What Can Go Wrong?

The global decline of brown algal forests along rocky coasts is causing an exceptional biodiversity loss. Regardless of conservation efforts, different techniques have been developed for large-scale restoration strategies in the Mediterranean Sea. In this study we tested ex situ pilot restoration of Gongolaria barbata (=Treptacantha barbata) for the first time in Slovenian coastal waters. Healthy apical fronds of the species were collected and the development of recruits on clay tiles was followed under laboratory conditions for 20 days. Despite the experimental difficulties experienced, especially due to the lack of antibiotics to prevent the growth of the biofilm, G. barbata recruits were outplanted in the sea on two concrete plates with 48 tiles each, protected by purpose-built cages to avoid grazing by herbivorous fish. The high survival rate of juveniles after four months in the field (89% of the tiles on the plate that was constantly protected) suggests that outplanting G. barbata is an operable approach for restoration efforts in the northern Adriatic Sea. Our first experiment in Slovenian coastal waters provides new information for the optimization of the best practices during the laboratory cultivation and addresses the early steps of restoration and introduction of young thalli in the natural environment.

Isolation of Native Arbuscular Mycorrhizal Fungi within Young Thalli of the Liverwort Marchantia paleacea

Arbuscular mycorrhizal fungi (AMF) are a group of soil microorganisms that establish symbioses with most land plant species. “Root trap culture” generally has been used for isolating a single regenerated spore in order to establish a monospecific, native AMF line. Roots may be co-colonized with multiple AMF species; however, only a small portion of AMF within roots sporulate, and do so only under certain conditions. In this study, we tested whether young thalli (<2 mm) of the liverwort Marchantia paleacea harbour monospecific AMF, and can be used as a vegetative inoculant line. When M. paleacea gemmae were co-cultivated with roots obtained from the field, the young thalli were infected by AMF via rhizoids and formed arbuscules after 18 days post-sowing. Ribosomal DNA sequencing of the AMF-colonized thalli (mycothalli) revealed that they harboured phylogenetically diverse AMF; however, new gemmae sown around transplanted mycothalli showed evidence of colonization from phylogenetically uniform Rhizophagus species. Of note, mycothalli can also be used as an inoculum. These results suggest that the young thalli of M. paleacea can potentially isolate monospecific AMF from field soil in a spore-independent manner.

Dynamics of Evernia prunastri (L.) Ach. and Hypogymnia physodes (L.) Nyl. population structure on Tilia cordata Mill. in the flooded lime-tree forest of the Bolshaya Kokshaga river

The paper deals with perennial dynamics of the population structure of the epiphytic lichens Evernia prunastri (L.) Ach. (2008-2016) and Hypogymnia physodes (L.) Nyl. (2010-2016) with the different life forms and the ecological strategies. The research was carried out in the Mari El Republic in the flooded lime-tree forest. The authors studied the density (number of thalli on a tree) on the trunk from 0 to 1,5 meters, the size and the ontogenetic structure of population of both species of different ontogenetic states: virginile (v1, v2), potentially generative (g1v, g2v, g3v), subsenile (ss) on 17 trees of Tilia cordata. The density of E. prunastri and H. physodes population increases. These changes are more specific for E. prunastri . Thalli of E. prunastri have a bigger size than thalli of H. physodes . These differences are observed on g1v, g2v, g3v, ss thalli. The decrease of thalli size is observed for both species but the difference is more for E. prunastri . The comparison of ontogenetic spectra of populations was made according to their heterogeneity within trees. The values of the effects (the effect is the parameter of the ontogenetic spectrum of the population) decreased in 2016, which indicated a shift of the ontogenetic spectra to the left, i.e. an increase in the proportion of young thalli that can be traced in the total ontogenetic spectra of populations. At the same time, the differences between the ontogenetic spectra of the E. prunastri population are statistically significant.

In vitro conservation strategy for endemic and endangered Himalayan liverwort Stephensoniella brevipedunculata Kashyap (Marchantiophyta)

During the present study an effort has been made to propagate (in-vitro) the endangered and endemic Himalayan liverwort Stephensoniella brevipedunculata Kashyap using different culture media under controlled Laboratory conditions. Axenic cultures of the taxon have been established using tubers as explants. Seven combinations of media with Full Strength Knop’s macronutrients; Half-strength Knop’s macronutrients; Half-strength Knop’s macronutrients + Vitamins; Halfstrength Knop’s macronutrients + 0.2 mg L-1 IBA + 0.1 mg L-1 BAP; Half-strength Knop’s macronutrients + 0.1 mg L-1 Kinetin + 0.1 mg L-1 2,4D; Half-strength Knop’s macronutrients + 0.1 mg L-1 IBA + 0.2 mg L-1 BAP and Hoagland no. 2 basal salt mixture were used for culture. The best growth was observed in the Hoagland no. 2 basal salt mixture medium, in which dichotomously branched young thalli were successfully formed. Subsequently healthy population of culture grown plants has been raised on soil in pots for the first time.

An Observation on Morphogenetic Response of Marchantia polymorpha subsp. ruderalis Bischl. and Boissel.-Dub. in Different Culture Media

The growth pattern of Marchantia polymorpha subsp. ruderalis, using gemmae as explant, in different culture media in combination with auxin and cytokinins has been studied. The plant growth was maximum in ½ KNOP's followed by KNOP's, Hoagland and Murashige and Skoog media. The normal growth of gemmae was inhibited at higher concentrations of hormone (1 mg L-1 Kinetin + 1 mg L-1 Auxin, 1.5 mg L-1 Kinetin + 1.5 mg L-1 Auxin, 0.5 mg L-1 2,4D) and callus like tissue was produced, while at lower concentrations (0.1 mg L-1 Kinetin+0.1 mg L-1 Auxin) gemmae developed normally and rhizoids were profusely produced on the surface of young thalli. Cultured thalli were transferred to the soil in pots and introduced in moss house.

Contribution to the lichen biota of the Stawy Milickie nature reserve and its adjacent area (Lower Silesia, southwestern Poland)

The paper presents the results of lichenological investigations conducted in the eastern part of the Stawy Milickie nature reserve and its buffer zone (Barycz Valley, southwestern Poland). The study area is a famous waterfowl refuge, consisting of several fishpond complexes, together with adjacent forests, meadows, and villages. In total 72 lichen taxa were recorded, growing on bark and branches of trees and shrubs, stumps, wood, soil, and anthropogenic rock substrates (mortar, concrete, bricks, etc.). Six species, namely <em>Evernia prunastri</em>, <em>Flavoparmelia caperata</em>, <em>Hypogymnia tubulosa</em>, <em>Parmelina tiliacea</em>, <em>Physconia distorta</em>, and <em>Pleurosticta acetabulum</em>, are threatened in Poland. As represented by single young thalli, they indicate the recent improvement of environmental conditions. The lichen biota of the study area is typical for the lowland regions of western and southwestern Poland.

New morphological aspects of cephalodium formation in the lichen Lobaria pulmonaria (Lecanorales, Ascomycota)

Cephalodia were investigated on young and mature thalli of Lobaria pulmonaria. Cephalodia originate from contact between hyphae and cyanobacteria on the upper or lower cortex or, less frequently, in the apical zone. Young thalli were found to associate with cyanobacteria even in the anchoring zone. Cephalodia formed on the young thalli or the anchoring hyphae share the same phenotypic characteristics. In spite of being composed of paraplectenchymatous hyphae, the cortex of mature thalli preserves a considerable plasticity, enabling the formation of cephalodia. The cyanobacterial incorporation process begins with cortical hyphae growing out towards adjacent cyanobacterial colonies, enveloping them and incorporating them into the thallus. The incorporation process is the same on the upper and the lower cortex. Early stages of cephalodia are usually found in young lobes, whereas in the older parts of the thallus only mature cephalodia are found.

Ulva flexuosa subsp. pilifera (Chlorophyta, Ulvophyceae) from the Wielkopolska region (West Poland): a new observation on the ultrastructure of vegetative cells

Ulva flexuosa subsp. pilifera previously known from northern Poland, from the channel near Szczecin and ponds near Łódź, has recently been found in the Malta Reservoir in the Wielkopolska (West Poland) region. Specimens collected in the Wielkopolska region were examined in detail, also under a transmission electron microscope (TEM). The morphometric analysis of Ulva thalli (both young and mature specimens) was performed in order to study the differences in the ultrastructure of vegetative cells. Rectangular cells in young thalli measured from 32.21–55.81 μm to 20.24–35.12 μm, and they formed clear longitudinal rows, while cells in the mature specimens ranged from 25.09–47.66 μm to 18.90–31.56 μm. This study indicates that vegetative cells of the mature thalli show tendency towards distortions of both the longitudinal and transverse cells arrangement. This distortion is determined by the development of possible carbonate calcium crystals on the thalli surface. The ultrastructural analysis (TEM) confirmed that the structure and placement of thylakoids is genus/species specific.