Moist Chamber Technique Research Articles

Myxobiota of the İskenderun Gulf (Mediterranean Sea/Türkiye) and its Environment

This study aims to determine the myxobiota of the İskenderun Gulf and its environment. This study was carried out on samples collected from 9 different stations in the İskenderun Gulf district in 2019-2022. Myxomycetes samples were collected from leaves, tree bark, and decayed and non-rotting plant materials. This aim is to detect both the myxobiota of this region and the halophytic myxomycetes species. The collected samples have developed myxomycete sporophores by using the moist chamber technique. In addition, myxomycetes which were developed in their natural environment were collected. As a result of the field and laboratory studies, 161 samples from different localities were collected and processed and 111 myxomycete samples were obtained from these samples. 41 species belonging to 6 ordo, 12 families, and 20 genera have been defined. 3 species were obtained only from the natural environment, 30 species were obtained only from moist chamber culture, and 8 species were obtained from both moist chamber culture and natural area. Arcyria cinerea, Didymium difforme, D. squamulosum, and Physarum pusillum were determined as abundant. While the number of species is rare in the seaside regions, it was observed that the further away from the sea, the number of species increased, especially in forest areas. A. cinerea, D. difforme, and D. dubium are the most common species on the beach and near the seaside. While cosmopolitan species were observed in abundance, Physarida members were found to be common in this results area. This study has contributed to the myxobiota of Türkiye.

A new species of Physarum (Physaraceae) from Turkey

A new species of Physarum, identified herein as P. karamanicum, developed on debris branches of Cedrus libani collected from the Sarıveliler district of Karaman province in Turkey and applied a moist chamber technique. P. karamanicum is morphologically similar to P. globuliferum, P. auripigmentum, and P. tenerum but is distinguished by sporotheca, stalk, capillitium, and spore features. The new species is characterized by a greenish-gray, globose sporotheca, light brown at the bottom, the remainder yellowish-cream colored stalk, triangular, convex quadrilateral nodes and pale-yellow capillitium, prominent spiny and groups of blunt warts spores, greenish gray peridium. Description of the new species, photographs (light microscope, stereo microscope, and scanning electron microscope), and a key to the closest morphologically related species of the genus are provided.

Results of myxomycete experiments could be affected by temperature in “standard” laboratory conditions

Moist chamber cultures are valuable for myxomycete research and can be used for the analysis of ecological patterns with implications in ecosystem conservation programs. However, to make comparisons between datasets valid, the method should be redesigned considering potential biases affecting the generation of results. In the present study, both the effect of the general climate of the laboratory and two microclimatic variables within the moist chamber were studied in relation with the obtained data. Of all the recorded variables, temperature was observed to affect the results, both at the level of the general laboratory climate and in relation with the microclimate of the moist chamber. Increments in laboratory and moist chamber temperature increased the probability, three or fourfold, associated with a higher number of records or species within a group of equivalent samples. Such probabilistic differences are significant enough to suggest that using the moist chamber technique in “standard” laboratory conditions is not enough for cross-dataset comparisons that increase the potential of myxomycete data for applications outside of the biological sciences.

Myxomycetes diversity in Adana Province (Turkey) with two new records

Samples were collected in Adana province during 2019–2021 and myxomycetes were cultured using the moist chamber technique. The samples consisted of rotten or live tree bark, leaves, rashes, vegetable, and animal material, which were considered likely to contain myxomycete spores. In adddition, natural myxomycete samples were collected at nine different stations. These ‘natural samples’ were immediately dried and potential spore-bearing material was kept in a warm and humid environment with the moist chamber technique. In total, 56 myxomycetes (205 samples; 148 by moist chamber technique and 57 ‘natural samples’) were obtained representing six orders, 12 families, 24 genera and 56 species. Eight species were collected only in natural samples, 35 species were obtained only by moist chamber technique in the laboratory, and 13 species were obtained by both methods. Two myxomycetes as new records (Licea rugosa and Physarum nivale) and one new locality (Cribraria lepida) were identified in Turkey

Konya-Beyşehir karayolu güzergahındaki miksomiset çeşitliliği

In this study, it was aimed to determine myxomycetes growing on materials collected from forests on Konya-Beyşehir (Turkey) highway route between 2019-2020. 253 materials such as log and stump materials, forest debris and bark of living tree were collected during the fieldworks in the region. The moist chamber technique was applied to the collected materials. As a result, 80 myxomycete specimens were developed and 21 myxomycete taxa belonging to 8 families were identified. The most common species is Perichaena depressa Lib. and was detected on 17 different substrates. In addition, the localities of the species (station number, substrate, collection date, collection number), and photographs of the species identified from the region are also given.

Cribraria lepida, Physarum dictyosporum, P. diderma, and P. spectabile newly recorded from Turkey

The moist chamber technique was used to obtain Cribraria lepida, Physarum dictyosporum, and P. diderma from wood or bark at different outdoor localities in Karaman Province and P. spectabile in Konya Province. These taxa are reported for the first time from Turkey.

Myxomycetes associated with three different types of Neotropical urban landscapes

Urban ecology is a research field gaining momentum due to contextual pressures to study macroecological effects on biodiversity. Herein, we evaluated the occurrence of myxomycetes in three urban Neotropical environments representing different levels of conurbation within different life zones. We used the moist chamber technique set up with ground litter collected by public roads for such evaluation. Results showed that 79% of the 420 moist chambers yielded myxomycetes and the same number of samples generated 570 records. The highest number of records and species were found in the cornurbated widespread city of the premontane moist forest life zone; and fewer records and species were recorded in the two non-conurbated compact cities. The lowest number of records was associated with the lowland dry forest life zone, whereas the lowest number of species was recorded in the desertic context. In general, more records and more species were observed in the outer zones relative to the urban centre but no significant differences were observed in either case. It is unlikely that the life zone characteristics could have primarily determined the results and highly likely played a secondary role. Also, previous geographical analyses in cities have reported no differences in myxomycete abundance or species richness in relation to the central parts of the urban landscape. The results presented herein support the latter idea, but also suggest that the presence of inter-conurbated green spaces and availability of substrates might play a role in the occurrence of sporocarps of myxomycetes in urban environments.

Pig (Sus scrofa domesticus) dung and their associated fungi: Potential candidates for bio-remediation and nutrients improvement of crude oil contaminated soils

Abstract. Enyiukwu DN, Bassey IN, Nwaogu GA, Asamudo NU, Chukwu LA, Monday EA, Maranzu JO. 2021. Pig (Sus scrofa domesticus) dung and their associated fungi: Potential candidates for bio-remediation and nutrients improvement of crude oil contaminated soils. Biodiversitas 22: 3276-3286. Pig dung a common agro-waste material in Nigeria is associated with many fungal saprophytes. Rising oil spillages and pollution in Nigeria have made cultivation of affected agro-lands un-rewarding. Potentials of pig dung in mitigating crude oil pollution in agro-soils are herein studied. The objectives of this study were to characterize the fungal profile associated with wet and dry dung of pig. And to evaluate the remediating impacts of pig dung and their associated fungi on crude oil and heavy metals contents, physicochemical properties, and phytotoxicity of crude oil polluted soils. Coprophilous fungi were isolated from wet and dry dung by moist chamber technique and identified. Utisoil from Umudike was polluted with crude oil, amended with 10-40% of dry pig dung and sown with soybean (Glycine max L.). The amended and un-amended crude oil contaminated utisols were analyzed to reduce crude oil and heavy metals contents; and physicochemical properties at planting, and harvesting, respectively, using classical and spectrometric methods, and the extent of crude oil and heavy metals removal determined. The percentage germination, height, number of leaves, leaf color, vigor, and biomass of the test crop were recorded from the test plant. The results showed that Aspergillus candidus, Sclerotium sp., Absidia sp., Acremonium sp., Candida tropicalis, C. pseudotropicalis, and Rhizopus oryzae were consistently isolated from the dry dung specimen than the wet one. The results also indicated that the treatments significantly reduced the treated soils' crude oil and heavy metals contents. It revealed that values obtained for residual oil or heavy metals contents of the treated soils and crop performance were inversely and directly related to levels of soil amendment respectively. Crop performance in the amended soil remained statistically (P?0.05) higher compared to the control. Soil analyses showed that the dung treatments significantly increased the values of nitrogen, phosphorus, potassium, organic carbon, etc. in a directly proportional manner. Therefore, pig dung and their associated fungi hold strong potentials as cheap eco-friendly platforms to provide and stimulate microbial growth, hasten degradation and removal of crude oil and heavy metals from contaminated soils; and thus permit and sustain good performance of arable crops.

Ecological assemblages of corticulous myxomycetes in forest communities of the North-East Ukraine

Corticulous myxomycetes remain one of the least surveyed ecological groups of terrestrial protists. These organisms develop on the bark of trees, mostly feeding on bacteria and microalgae. Their microscopic size and fast developmental cycle (3–5 days) complicate the study of these organisms, and therefore data their on ecological relationships and patterns of biodiversity corticulous myxomycetes remain controversial. On the territory of the southwest spurs of the Central Russian Upland (Northeast Ukraine), no special studies on these organisms have been conducted. During 2017–2020, in nine forest sites located in this territory, we collected samples of bark of 16 species of tree plants, on which sporulating myxomycetes were then identified using the moist chamber technique in laboratory conditions. A total of 434 moist chambers was prepared, and 267 (61.5%) of which were found to contain myxomycete fruiting bodies. In total, we made 535 observations, finding 20,211 sporocarps. As a result, in the surveyed territory, we found 38 species of corticulous myxomycetes, belonging to 18 genera, 10 families, 7 orders, and 2 subclasses of Myxomycetes. Among the species of corticulous myxomycetes, the most abundant were Echinostelium minutum, Arcyria pomiformis, Macbrideola cornea, Perichaena chrysosperma, Licea kleistobolus, Paradiacheopsis fimbriata, Cribraria violacea, Enerthenema papillatum, A. cinerea, and L. operculata. The greatest species richness in the examined biota was observed for genera Comatricha, Licea, Paradiacheopsis and Perichaena, families Amaurochaetaceae and Trichiaceae, orders Stemonitidales, Trichiales and Physarales. By species diversity, dark-spored myxomycetes (Collumellomycetidae) somewhat exceeded bright-spored myxomycetes (Lucisporomycetidae). Badhamia versicolor, Didymium dubium, D. sturgisii, Macbrideola decapillata, and Perichaena luteola are new species for the surveyed area. Four species of myxomycetes were collected in Ukraine for the first time: Hemitrichia pardina, Licea floriformis, L. pygmea, and Macbrideola argentea. Quantitative and qualitative structure of myxomycete consortia developing on different species of substrate-forming plants demonstrated significant differences. The highest level of similarity was demonstrated by Fraxinus excelsior and Acer platanoides, and a relatively strong relationship was seen between Pinus sylvestris and Tilia cordata. The central cluster comprised F. excelsior, A. platanoides and P. sylvestris. By the sum of values of Bray-Curtis coefficient, Quercus robur appeared to be most distinctive plant species by quantitative composition of myxomycete consortia. F. excelsior and T. cordata are the most favourable for the development of corticulous myxomycetes. In all the analyzed consortia, the dominant species belonged to the Stemonitidales and Trichiales orders, while the remaining orders were represented by notably fewer species. Relative species richness of Stemonitidales was the highest in consortia of P. sylvestris, the contribution of Liceales was the greatest in A. platanoides and P. sylvestris, the percentage of Echinosteliales and Physarales was the highest on F. excelsior, the share of Cribrariales was especially large on A. platanoides. Trichiales were represented on all the analyzed substrates to almost the same extent. Representatives of Cribrariales and Physarales were completely absent on P. sylvestris, the species of Clastodermatales – on all species of plants, except Q. robur. Prevalence of bright-spored myxomycetes was determined for consortia of Acer platanoides, the dominance of dark-spored myxomycetes – for F. excelsior, P. sylvestris and Q. robur. The obtained data indicate the presence of stable complexes of corticulous myxomycetes, associated with different species of trees in the forest ecosystems of Northeast Ukraine. This encourages further study of the structure of myxomycete consortia with tree species that were not included in this study and determining the influence of physical-chemical properties of the bark of different plant species on the discovered peculiarities of myxomycete communities.

Myxomycetes within ecotones in temperate and tropical forests

Ecotones are interesting portions of forests where microorganisms can be studied for monitoring and diversity purposes. Given the faster life cycle of these organisms, their study in such a context is important to develop models that allow us to understand adaptive microbial dynamics. The main objective of this study was to evaluate the dynamics of myxomycetes present in the leaf litter layer in different geographical locations representing both temperate and tropical systems, using an experimental approach within the ecotones produced by sharp forest edges. Employing the moist chamber technique and three ecological indicators (species richness, diversity, and number of records, as a proxy for abundance) associated with the reproductive phase of the life cycle of myxomycetes, the present study summarized the potential effect of both temperate and tropical ecotones on the distribution of these organisms. Data from 26 locations in six countries showed that all indicators were higher in temperate than in tropical forests and that most differences could be explained by ecological effects. The overall influence of the ecotone on myxomycetes was opposite in temperate and tropical forests, but the data collected for both types of forests showed similarities that could be explained by comparable mathematical models. These results suggested that, with the methods used herein, ecotones do affect ecological patterns of myxomycetes, but the intensity and directionality of such an effect differ depending on the forest type involved.

<strong>Myxomycetes biodiversity in Gaziantep Province (Turkey) with four new records</strong>

Myxomycetes were cultured in moist chambers using substrate material collected in Gaziantep province, Turkey, during 2017–2019. Fruit bodies of wild myxomycetes were collected at ten locations. Rotten or live tree bark, leaves, debris, vegetable, and animal material, which were considered likely to contain spores, were also collected. Natural samples were immediately dried, and potential spore-bearing material was kept in a warm and humid environment with the moist chamber technique. A total of 537 samples were studied and 203 myxomycetes isolates were obtained, 33 of which were natural samples, 76 were obtained with the moist chamber technique and 94 were obtained both naturally and with the moist chamber technique in the laboratory. Six orders, 9 families, 16 genera and 42 species were identified in 3 subclasses. All species were new in Gaziantep province and four myxomycetes were identified as new records in Turkey; Didymium atrichum Henney & Alexop., Didymium serpula Fr., Craterium obovatum Peck and Physarum bivalve Pers. were added to the Turkish mycota.

Myxomycete diversity on Cryptomeria japonica bark varies with land-use type along montane rivers

Myxomycetes occur globally, but little is known about the mechanism by which myxomycete diversity and community structure respond to environmental gradients and human activity at local scales. The present study assessed the distribution of corticolous myxomycetes living on the bark of <em>Cryptomeria japonica</em> trees along three rivers originating in the Chugoku Mountains in western Japan. Bark samples were collected from 14 sites along each river, which encompass the river sources as well as the upper, middle, and lower reaches. The environmental characteristics of each site were assessed for three variable types: geography, climate, and land-use. A 1 km2 grid was superimposed onto each survey site, consisting of 100 cells of 1 ha each, and each cell was classified using aerial imagery into a broad land-use type. Of these, three land-use types (forest, farmland, and residential) were regularly used as a comprehensive indicator of landscape. The bark samples were cultured using the moist chamber technique, and the resulting myxomycete fruiting bodies were identified. Sporophores formed on 96% of the 1,490 moist chamber cultures and were classified into 27 taxa (26 species and one variety). Species diversity was highest in natural forests near river sources and decreased with proximity to downstream estuaries. Community similarities between survey sites were analysed using non-metric multidimensional scaling (NMDS). The first NMDS axis was negatively correlated with distance from estuary and forest coverage. Indicator species were identified for environmental changes along river basins. The relative abundances of <em>Macbrideola argentea</em>, <em>Hemitrichia velutina</em>, and <em>Physarum nutans</em> var. <em>rubrum</em> were positively correlated with forest coverage. In contrast, the relative abundances of <em>Clastoderma debaryanum</em>, <em>Diderma chondrioderma</em>, and <em>Echinostelium minutum</em> were positively correlated with residential area coverage. The distribution of corticolous myxomycetes on C. japonica trees was associated with local landscape changes along the river environments. This is the first report on myxomycetes assemblages along river basins (acting as ecological corridors) and indicates that forest degradation and land-use types strongly affect myxomycete diversity on the bark of living <em>C. japonica</em> trees.

Myxomycetes Diversity of Belen Region of Hatay Province (Turkey)

Aim of study: The present study aimed to determine myxomycetes diversity in Belen region located in Hatay province, Turkey. Area of study: The present study was conducted on samples collected in 10 stations in Belen district during 2012 and 2013. Material and methods: Myxomycete sporophores were collected from leaf, tree bark, decayed and non-rotting plant material. Myxomycete sporophores were developed with the moist chamber technique in collected samples. Further myxomycete sporophores were collected in their natural environment. Main results: As a result of the field and laboratory studies, 40 species in 9 families and 19 genera were identified. Highlights: The present study contributed to Turkish Myxobiota.

Discovery of Three New Phytotoxins from the Fungus Aspergillus nidulans by Pathway Inactivation.

Fungi are a source of novel phytotoxic compounds to be explored in the search for effective and environmentally safe herbicides. The genetic inactivation of the biosynthetic pathway of the new phytotoxin cichorine has led to the isolation of three novel phytotoxins from the fungus Aspergillus nidulans: 8-methoxycichorine (4), 8-epi-methoxycichorine (5), and N-(4’-carboxybutyl) cichorine (6). The structure of the new compounds was clearly determined by a combination of nuclear magnetic resonance (NMR) analysis and high-resolution electrospray ionization (HRESIMS). The phytotoxic bioassay was studied on leaves from Zea mays and Medicago polymorpha L. at the concentration of 5 × 10−3 M by using a moist chamber technique. Novel phytotoxins 8-methoxycichorine (4), 8-epi-methoxycichorine (5), and N-(4’-carboxybutyl) cichorine (6) exhibited a better phytotoxic effect than cichorine.

Australian corticolous myxomycetes: models of distribution and development

This paper presents an integrated model of the variation over a continental landmass of myxomycetes, a single-celled organism in the phylum Amoebozoa. Bark samples were collected on long traverses across Australia, and cultivated in Petri dishes by the moist chamber technique to obtain large assemblages of common species. The results of this survey and previous surveys are consistent with there being four major myxomycete assemblages: Tropical, Northern Arid, Southern Arid and Temperate. Where mapped, these species assemblage regions are consistent with the Australian phytogeographical regions. The myxomycetes differ between arid and non-arid areas; the arid areas have slightly higher productivity per wetting event, with members of the Physarales and Liceales relatively important and the Stemonitidales, Trichiales and Cribrariales less important. When the bark samples are placed in a moist culture there is a myxomycete growth cycle and then the population declines to resting phases. The population increase during a growth phase can be modelled by a linear plot of log(abundance) against species rank, where abundance is total harvested spore volume of a species. The population decline appears to be linear from two weeks after watering, declining to negligible activity 4 weeks after watering.

Some Mycetozoa (Myxomycetes) Members from Zorkun High Plateau (Osmaniye)

In this study, Myxomycetes collected from 4 different locality from Zorkun Plateau (Osmaniye-Turkey) in 2014-2016. As a result of field and laboratory studies in two years we reported 28 species belonging to 9 family and 16 genus. 7 species collected only natural area, 16 species obtained Moist Chamber Technique in laboratory, 5 species obtained both natural and Moist Chamber Technique. Echinostelium minutum, Cribraria cancellata, Didymium difforme, Didymium squamulosum, Physarum album , and Arcyria cinerea are the most common species. 6 species are abundant, 10 species common and 12 species occasional, and species/genus ratio (S/G) is 1.75. Biodiversity, distribution, habitat characteristics and numbers of species are discussed in details.

Does elevation influence the distributional patterns of tropical myxomycetes? A case study in Costa Rica

ABSTRACTIn order to test the hypothesis that elevation may be an important factor accounting for the distribution of myxomycetes in tropical forests, this project was designed and conducted in Costa Rica. Two lower elevational belts were selected for this work due to their floristic and structural resemblance. Using the moist chamber technique, 40 different sites located in four different transects in two different macroclimatic regions were surveyed using three substrates during the rainy and the dry periods of 2014 and 2015. The results showed a lack of differences in diversity-based estimators according to elevation using different approaches and taxonomic differences were found across transects, collecting periods and substrates but not in relation with elevation either. Our results suggest that when increased spatial sampling resolution is implemented and floristic elements are common, elevation may not be as important of a factor in shaping the distribution of myxomycetes in tropical forests as commonly believed.

Microfungi Rhizo-Pyllosphere, Endomycorrhizae of Banana (Musa paradisiaca L.) in Karangwangi Countryside, South Cianjur, West Java Indonesia

Fermented banana “sale pisang” are hold potential home industry in Karangwangi countryside, in South Cianjur. However, recently sale pisang’s production is affected significantly by the pathogenic fungi. Research about microfungi rhizo-phyllosphere and endomycorrhizae of banana plant (Musa paradisiaca L.) in West Java aims to identify this reason. Preliminary survey to research locations using explorative method, sampling soils around roots, and leaves. Microfungi culture and counting were conducted by dilution technique and Total Plate Count (TPC) methods. Fungi identification was performed with Moist Chamber Technique based on morphology. While plant determination surround of banana plant, infection rate and colouring of endomycorrhizae were conducted according to the standard protocol described elsewhere. In this study, 11 species of microfungi were identified, namely Acremonium sp., Cladosporium sp., Colletotrichum sp., Curvularia sp., Fusarium sp., Monilia sp., Nigrospora sp., Penicillium sp1., Penicillium sp2., Phytophthora sp. and Trichoderma sp. Eventually Endomycorrhizae species were Gigaspora sp. and Acaulospora sp. Infection rate of plants surrounding banana plant were varied for Mimosa pudica, Cyperus rotundus, Stachytarpheya jamaicinensis, Cyperus kyllingia, Melinis minutiflora, Erechtites hieracifolia, Arachis hypogaea, Momordica charantia, Euphorbia hyrta, Amaranthus gracilis, Manihot utillissima, Ageratum conyzoides, Emilia sonchifolia, Widelia triloba, Amaranthus spynosus and Solanum nigrum as much as 20, 20, 20, 40, 60, 80, 100, 20, 60, 40, 100, 40, 40, 40, 40 and 20%, respectively. This species-specific rate of infection is discussed in order to obtain insight how the diversity of microfungi may affect mycorrhizal infection in banana plants.

A comparative species listing of myxomycetes from tropical (Philippines) and temperate (United States) forests

In terms of their flora and fauna, tropical forests are generally reported to be characterized by higher species diversity than temperate forests. Can this also be true for microorganisms such as myxomycetes? In the present study, three lowland tropical forests in the Philippines and three midlatitude temperate forests in north central Arkansas in the United States were surveyed for myxomycetes with the moist chamber technique as it applies to these organisms. Results indicated that more species of myxomycetes were associated with samples of aerial litter (dead but still attached plant parts), forest floor litter and woody twigs collected in Arkansas than for those collected in the Philippines. A higher value for taxonomic diversity also was noted for the temperate forests. However, 19 species listed herein are new records for the Philippines. In the present study, a comparison of the taxonomic diversity of myxomycetes in relation to the number of moist chamber replications was carried out, and this showed that a difference of as many as eight species could occur between what was recorded from a single culture and a series of three cultures prepared from the same sample. Clearly, many of these species could be missed if only a single culture is prepared for a particular sample.

Isolation, structural identification and herbicidal activity of N-phenylpropanamide from Streptomyces sp. KA1-3

The secondary metabolites produced by Streptomyces sp. KA1-3, cultured on starch casein broth, was extracted by ethyl acetate and concentrated. Purification of the compound by thin layer chromatography lead to isolation of N-phenylpropanamide from one polar fraction. The structure of the herbicidal compound was elucidated on the basis of UV, FT-IR, mass and H1 NMR spectroscopy. The herbicidal activity of the isolate was tested against the weeds Cassia occidentalis L. and rhizome Cyperus rotundus L. by moist chamber technique and rolled towel paper assay method. Herbicidal activity of the bioactive compound N-phenylpropanamide was further evaluated under in vitro condition. The herbicidal compound showed 80% of seed germination inhibition in C. occidentalis L. and rhizome C. rotundus L. weed. The actinobacterium can be used as a source for bioherbicidal agent.