Fungal Diversity in Deep Sea Sediments from East Yap Trench and Their Denitrification Potential

Abstract

In recent years, the hadal trenches have been recognized as biological hot spots for deep sea researchers. Due to high hydrostatic pressure, low temperatures, high salinity and low nutrients, the microorganisms in hadal trenches may have unique community structure with potential for biotechnical application. Compared with bacteria and archaea, the diversity and ecological roles of fungi in hadal trenches remain largely unknown. The purpose of this study was to explore fungal diversity in deep-sea sediments of the Yap trench and their denitrification potential. In the present study, a total of 106 fungal strains were isolated from six sediment samples collected in the East Yap Trench. These fungi belonged to five classes (Dothideomycetes, Eurotiomycetes, Sordariomycetes, Cystobasidiomycetes, and Microbotryomycetes), thirteen genera (Acremonium, Alternaria, Aureobasidium, Aspergillus, Cladosporium, Cystobasidium, Engyodontium, Gliomastix, Lecanicillium, Penicillium, Phoma, Rhodotorula and Trichoderma) and eighteen species, based on morphological identification and ITS-rDNA sequence analysis. Among them, the dominant genus is Cladosporium, which accounting for 42.45% of the total fungal strains. Meanwhile, the denitrification potential of the fungal strains was also examined with two different denitrifying media (nitrate and nitrite as sole substrate, respectively). Two fungal strains (Acremonium sp. and Aspergillus versicolor), were found to be able to produce N₂O ex situ in the presence of nitrite. No fungus was found to produce N₂O by using nitrate. Our results suggest that fungi in hadal sediments, play important roles in nitrogen cycles.     

Diversity of culturable filamentous Ascomycetes in the eastern South Pacific Ocean off Chile

Our study reports the diversity of culturable mycoplankton in the eastern South Pacific Ocean off Chile to contribute with novel knowledge on taxonomy of filamentous fungi isolated from distinct physicochemical and biological marine environments. We characterized spatial distribution of isolates, evaluated their viability and assessed the influence of organic substrate availability on fungal development. Thirty-nine Operational Taxonomic Units were identified from 99 fungal strains isolated from coastal and oceanic waters by using Automatic Barcode Gap Discovery. All Operational Taxonomic Units belonged to phylum Ascomycota and orders Eurotiales, Dothideales, Sordariales and Hypocreales, mainly Penicillium sp. (82%); 11 sequences did not match existing species in GenBank, suggesting occurrence of novel fungal taxa. Our results suggest that fungal communities in the South Pacific Ocean off Chile appear to thrive in a wide range of environmental conditions in the ocean and that substrate availability may be a factor influencing fungal viability in the ocean.     

Distribution of the antifungal agents sordarins across filamentous fungi

Sordarins are a class of natural antifungal agents which act by specifically inhibiting fungal protein synthesis through their interaction with the elongation factor 2, EF2. A number of natural sordarins produced by diverse fungi of different classes have been reported in the literature. We have run an exhaustive search of sordarin-producing fungi using two different approaches consecutively, the first one being a differential sensitivity screen using a sordarin-resistant mutant yeast strain run in parallel with a wild type strain, and the second one an empiric screen against Candida albicans followed by early detection of sordarins by LC–MS analysis. Using these two strategies we have detected as many as 22 new strains producing a number of different sordarin analogues, either known (sordarin, xylarin, zofimarin) or novel (isozofimarin and 4′-O-demethyl sordarin). Sordarin and xylarin were the most frequently found compounds in the class. The producing strains were subjected to sequencing of the ITS region to determine their phylogenetic affinities. All the strains were shown to belong to the Xylariales, being distributed across three families in this order, the Xylariaceae, the Amphisphaeriaceae, and the Diatrypaceae. Despite being screened in large numbers, we did not find sordarin production in any other fungal group, including those orders where sordarin producing fungi are known to exist (i.e., Sordariales, Eurotiales, and Microascales), suggesting that the production of sordarin is a trait more frequently associated to members of the Xylariales than to any other fungal order.     

Fungal load in <Emphasis Type="Italic">Bradysia agrestis</Emphasis>, a phytopathogen-transmitting insect vector

Larvae of Bradysia agrestis, a phytopathogen-transmitting insect vector in East Asia, were sampled from geographically (ecologically) segregated regions to identify their intestinal fungal flora. A total of 24 fungal strains were isolated from the insect vectors and selected based on morphological differences. In addition, 38 fungal strains were isolated from the ulcerated parts of invaded host plants by the same method, revealing the impact of vector fungal flora on their host plants. For molecular identification of the fungi, internal transcribed spacer (ITS) regions were amplified and sequenced. Their sequences were compared with sequences of other fungal strains obtained from NCBI GenBank, and their phylogeny was determined. The dominant fungal genera in the insect vector were Penicillium (25%), Aspergillus (21%), and Cladosporium (13%). In plant scar lesions, most fungal isolates belonged to the genera Fusarium (31.6%), Phoma (7.8%), Didymella (7.8%), and Epicoccum (7.8%). Fungal genera in vectors or host plant lesions differed by study site. Furthermore, diversity indices by study site showed clear differences based on Margalef’s richness (2.06, 2.40, 3.04), and Menhinick’s (1.89, 2.12, 2.53), and Simpson’s indices (0.14, 0.07, 0.07). In addition, common fungal strains in insect vectors were found to be closely related to members of the genera Cladosporium, Penicillium, or Aspergillus. Among these strains, those showing the highest homology with Aspergillus terreus, which regarded as beneficial fungal genera could be considered ideal paratransgenesis candidates. Some other fungal strains from vectors or ulcerated plant parts from each study site after B. agrestis invasion may be harmful in terms of plant disease or agrifood safety. This study provides information on the fungal microbiota of B. agrestis, an emerging problem in East Asia, and proposes paratransgenesis candidates to control this insect vector. Furthermore, potential transferable pathogens or commensal fungi were revealed by comparing the fungal biota between the insect gut and the ulcerated parts of the invaded host plants.     

Susceptibility of phosphogypsum to fungal growth and the effect of various biocides

Natural gypsum (NG) and phosphogypsum (PG) were tested for resistance to fungal growth based on standard test ASTM D 3273-86, with the recommended mixture of three fungal species, and using the same test modified by the use of a Cladosporium sp. A, isolated from a gypsum plaster ceiling. In the standard test little growth occurred on any of the test specimens. However, abundant fungal growth was produced by the Cladosporium sp. A on phosphogypsum, which was much more susceptible than natural gypsum. Phosphogypsum heated to 600°C to destroy organic residues was resistant to growth of Cladosporium sp. A, as well as other fungi isolated from phosphogypsum panels stored in the environment for 2 years: Cladosporium sp. B, Aspergillus niger and Trichoderma sp. Phosphogypsum moulded in Petri dishes was susceptible to growth of a wide range of fungi, although Fusarium sp. and Rhizopus sp. caused practically no discoloration of the substrate. Six biocides were separately incorporated into the phosphogypsum at concentrations recommended by the suppliers and test specimens incubated on Sabouraud agar inoculated with various fungal isolates. The biocide 2-N-octyl-4-isothiazolin-3-one was the most efficient compound. It prevented the growth of the fungi most likely to cause health problems in buildings, but not that of Helminthosporium sp., isolated from powdered phosphogypsum in the factory. This was the most resistant fungus showing growth on all biocide-containing specimens.     

Culturable endophytic filamentous fungi from leaves of transgenic imidazolinone-tolerant sugarcane and its non-transgenic isolines

The diversity of endophytic filamentous fungi from leaves of transgenic imidazolinone-tolerant sugarcane plants and its isoline was evaluated by cultivation followed by amplified rDNA restriction analysis (ARDRA) of randomly selected strains. Transgenic and non-transgenic cultivars and their crop management (herbicide application or manual weed control) were used to assess the possible non-target effects of genetically modified sugarcane on the fungal endophytic community. A total of 14 ARDRA haplotypes were identified in the endophytic community of sugarcane. Internal transcribed spacer (ITS) sequencing revealed a rich community represented by 12 different families from the Ascomycota phylum. Some isolates had a high sequence similarity with genera that are common endophytes in tropical climates, such as Cladosporium, Epicoccum, Fusarium, Guignardia, Pestalotiopsis and Xylaria. Analysis of molecular variance indicated that fluctuations in fungal population were related to both transgenic plants and herbicide application. While herbicide applications quickly induced transient changes in the fungal community, transgenic plants induced slower changes that were maintained over time. These results represent the first draft on composition of endophytic filamentous fungi associated with sugarcane plants. They are an important step in understanding the possible effects of transgenic plants and their crop management on the fungal endophytic community.     

Detection and monitoring of anaerobic rumen fungi using an ARISA method

Aim: To develop an automated ribosomal intergenic spacer region analysis (ARISA) method for the detection of anaerobic rumen fungi and also to demonstrate utility of the technique to monitor colonization and persistence of fungi, and diet‐induced changes in community structure. Methods and Results: The method could discriminate between three genera of anaerobic rumen fungal isolates, representing Orpinomyces, Piromyces and Neocallimastix species. Changes in anaerobic fungal composition were observed between animals fed a high‐fibre diet compared with a grain‐based diet. ARISA analysis of rumen samples from animals on grain showed a decrease in fungal diversity with a dominance of Orpinomyces and Piromyces spp. Clustering analysis of ARISA profile patterns grouped animals based on diet. A single strain of Orpinomyces was dosed into a cow and was detectable within the rumen fungal population for several weeks afterwards. Conclusions: The ARISA technique was capable of discriminating between pure cultures at the genus level. Diet composition has a significant influence on the diversity of anaerobic fungi in the rumen and the method can be used to monitor introduced strains. Significance and Impact of the Study: Through the use of ARISA analysis, a better understanding of the effect of diets on rumen anaerobic fungi populations is provided.     

Antimicrobial potentials of endophytic fungi residing in <Emphasis Type="Italic">Quercus variabilis</Emphasis> and brefeldin A obtained from <Emphasis Type="Italic">Cladosporium</Emphasis> sp.

Among 67 endophytic fungi isolated from Quercus variabilis, 53.7% of endophytic fungal fermentation broths displayed growth inhibition on at least one test microorganism, such as pathogenic fungi (Trichophyton rubrum, Candida albicans, Aspergillus niger, Epidermophyton floccosum, Microsporum canis) and bacteria (Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens). Moreover, 19.4% of strains showed a broader antimicrobial spectrum, such as Aspergillus sp., Penicillium sp., Alternaria sp., 20.9% of strains showed strong inhibition (+++) to pathogenic bacteria, while only 7.5% displayed that to test fungi. The most active antifungal strain I(R)9-2, Cladosporium sp. was selected and fermented. From the broth, a secondary metabolite, brefeldin A was obtained. This is the first report on the antimicrobial potentials of endophytic fungi residing in Q. variabilis and isolation of brefeldin A produced by Cladosporium sp.     

Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis

The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and β-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or β-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores.     

Denitrification activity of Bradyrhizobium sp. isolated from Argentine soybean cultivated soils

Two hundred and fifty strains, all of them representatives of native Bradyrhizobium sp., isolated from soils cultivated with soybean have been characterized by their denitrification activity. In addition, the denitrification potential of those soils was also measured by evaluating the most-probable-number (MPN) of denitrifying bacteria and the denitrification enzyme assay (DEA). Of the 250 isolates tested, 73 were scored as probable denitrifiers by a preliminary screening method. Only 41 were considered denitrifiers because they produced gas bubbles in Durham tubes, cultures reached an absorbance of more than 0.1 and NO³⁻ and NO²⁻ were not present. Ten of these 41 were selected to confirm denitrification and to study denitrification genes. According to N₂O production and cell protein concentration with NO³⁻, the isolates could be differentiated in three categories of denitrifiers. The presence of the napA, nirK, norC and nosZ genes was detected by production of a diagnostic PCR product using specific primers. RFLP from the 16S-23S rDNA spacer region (IGS) revealed that denitrifiers strains could be characterized as Bradyrhizobium japonicum and strains which were non-respiratory denitrifiers as B. elkanii.     

The diversity,distribution, and pathogenic potential of cultivable fungi present in rocks from the South Shetlands archipelago,Maritime Antarctica

We studied the molecular taxonomy and diversity of cultivable rock fungi from Antarctic islands. From 50 rock samples, 386 fungal isolates were obtained and identified as 33 taxa of 20 genera. The genera Cladophialophora, Cladosporium, Cyphellophora, Eichleriella, Paracladophialophora, and Penicillium displayed the highest densities. Ecological diversity indices showed that the fungal assemblages are diverse and rich with low dominance. The genera Cladophialophora, Cladosporium, and Penicillium showed a broad distribution from rocks of the various islands. One hundred and fifty-nine fungi, grown at 37 °C, were identified as Penicillium chrysogenum, Fusarium sp., and Rhodotorula mucilaginosa. One hundred and three fungi displayed haemolytic activity, 81 produced proteinase, 9 produced phospholipase, and 25 presented dimorphism and a spore diameter ≤ 4 µm. The Antarctic Peninsula region appears to be under the effects of global climate changes, which may expose and accelerate the rock’s weathering processes, and expose and release cryptic fungi and other microbes, especially those with innate pathogenic potential, previously arrested in rocks. Consequently, these rocks and their particles may represent a vehicle for the dispersal of microbial propagules, including those able to spread pathogens, along, across, and out of Antarctica.

     

不同种植年限薰衣草根际土壤真菌群落结构的演变

土壤真菌群落结构和多样性对植物生长起着重要作用,了解种植年限对薰衣草根际土壤真菌群落结构的影响对薰衣草病害防治和增产的研究具有重要意义。探究不同种植年限薰衣草根际土壤真菌群落结构和多样性特征,及其随种植年限的演变规律。采集新疆伊犁霍城县种植年限1、3、5 a,以及未种植薰衣草土壤,对ITS序列进行Illumina高通量测序。对测序结果进行分析,比较各样品组真菌多样性和群落分布规律及与种植年限的关联。结果表明,Alpha多样性分析显示随着种植年限的增加Shannon指数逐渐降低,而Chao1指数先降低后增高。在种植薰衣草土壤中共检测到12个门,28个纲,72个目,146个科,236个属。在门水平上,子囊菌门(Ascomycota)、担子菌门(Basidiomycota)、球囊菌门(Glomeromycota)为优势菌门。随着种植年限的增加,子囊菌门相对丰度逐渐降低,担子菌门逐渐增高,而球囊菌门先增加后降低。在属水平上优势菌属为Xylodon、锐孔菌属(Oxyporus)、镰刀菌属(Fusarium)、枝孢属(Cladosporium),均属于植物致病菌。随着种植年限的增加,Xylodon...     

Diversity and Antimicrobial Activity of Culturable Endophytic Fungi Isolated from Moso Bamboo Seeds

Bamboos, regarded as therapeutic agents in ethnomedicine, have been used to inhibit inflammation and enhance natural immunity for a long time in Asia, and there are many bamboo associated fungi with medical and edible value. In the present study, a total of 350 fungal strains were isolated from the uncommon moso bamboo (Phyllostachys edulis) seeds for the first time. The molecular diversity of these endophytic fungi was investigated and bioactive compound producers were screened for the first time. All the fungal endophytes were categorized into 69 morphotypes according to culturable characteristics and their internal transcriber spacer (ITS) regions were analyzed by BLAST search with the NCBI database. The fungal isolates showed high diversity and were divided in Ascomycota (98.0%) and Basidiomycota (2.0%), including at least 19 genera in nine orders. Four particular genera were considered to be newly recorded bambusicolous fungi, including Leptosphaerulina, Simplicillium, Sebacina and an unknown genus in Basidiomycetes. Furthermore, inhibitory effects against clinical pathogens and phytopathogens were screened preliminarily and strains B09 (Cladosporium sp.), B34 (Curvularia sp.), B35 (undefined genus 1), B38 (Penicillium sp.) and zzz816 (Shiraia sp.) displayed broad-spectrum activity against clinical bacteria and yeasts by the agar diffusion method. The crude extracts of isolates B09, B34, B35, B38 and zzz816 under submerged fermentation, also demonstrated various levels of bioactivities against bambusicolous pathogenic fungi. This study is the first report on the antimicrobial activity of endophytic fungi associated with moso bamboo seeds, and the results show that they could be exploited as a potential source of bioactive compounds and plant defense activators. In addition, it is the first time that strains of Shiraia sp. have been isolated and cultured from moso bamboo seeds, and one of them (zzz816) could produce hypocrellin A at high yield, which is significantly different from the other strains published.     

Screening and characterization of antifungal clusterbean (Cyamopsis tetragonoloba) rhizobacteria

About 377 guar (Cyamopsis tetragonoloba) rhizobacteria were isolated from cultivated soils of north-west India (Thar Desert) and their antifungal activity against Macrophomina phaseolina (strains of groundnut, mungbean and guar) and Fusarium oxysporum (strains of chickpea and cumin) was examined. Isolates were characterised for generic types and physiological/functional diversity. About 19% isolates representing 24% locations were inhibitory to fungal growth. Isolates 009071, 009073, 009078 and 102354 recorded maximum inhibition of pathogenic fungi on plates. Isolate 034206 gave highest %RI, 009073 showed maximum protease activity and 102354 gave highest salt tolerance. Net house and field screening results revealed that isolates 004052, 009071, 009073, 001001, 094340 and 102354 had potential for biocontrol of disease. Partial sequencing of 16S rRNA gene of 61 isolates showed that 85% of isolates belonged to genus Bacillus. Phylogenetically, however, there were four clusters in the Bacillus group comprising of Bacillus subtilis, B. cereus, B. pumilus and B. sphaericus. One isolate was identified as B. flexus, while six isolates were Bacillus spp. Four isolates were identified as Achromobacter xylosoxidans, two as Bacterium (unclassified bacteria), and one each as Ochrobactrum intermedium, Pseudomonas aeruginosa and Ralstonia sp.     

NirK and nirS Nitrite reductase genes from non-agricultural forest soil bacteria in Thailand

The genetic heterogeneity of the nitrite reductase gene (nirK and nirS) fragments from denitrifying prokaryotes in a non-agricultural forest soil in Thailand was investigated using soil samples from the Plant Germplasm-Royal Initiation Project area in Kanchanaburi Province, Thailand. Soil bacteria were screened for denitrification activity and 13 (from 211) positive isolates were obtained and further evaluated for their ability to reduce nitrate and to accumulate or reduce nitrite. Three species with potentially previously unreported denitrifying activities were recorded. Analysis of the partial nirK and nirS sequences of these 13 strains revealed a diverse sequence heterogeneity in these two genes within the same environment and even potentially within the same host species, the potential existence of lateral gene transfer and the first record of both nirK and nirS homologues in one bacterial species. Finally, isolates of two species of bacteria (Corynebacterium propinquum and Micrococcus lylae) are recorded as denitrifiers for the first time.     

Effect of pre-harvest fungicides on the spoilage of soft fruit after harvest

In the 1972 season, Elvaron and Benlate reduced the count of Botrytis cinerea, Cladosporium spp., Penicillium spp. and Aureobasidiutn pullulans on strawberry and raspberry fruits during the first 2 wk of the season, but not subsequently. No such reduction was recorded for Hainesia lythri (strawberries only), Phoma state of Didymella applanata (raspberries only), Mucor spp., and Rhizopus spp. Similar results were obtained in 1973. In the first half of either season, both fungicides decreased the rate of fungal spoilage of stored fruit (except for Benlate in the 1973 season), although in the latter part of the season, there was little difference in the incidence of fungal spoilage between sprayed and unsprayed fruit. Both fungicides, however, reduced the development of B. cinerea (and Cladosporium spp. on raspberries) on stored fruit irrespective of season or harvest date except for Benlate in the 1973 season where Benlate-resistant strains of the fungi developed on the fruit. The failure of either fungicide to reduce fungal spoilage in the latter part of the harvesting season was due to the development of Phycomycetes, particularly Mucor mucedo and to a lesser extent Rhizopus stolonifer and R. sexualis, except where Benlate-resistant strains of B. cinerea and Cladosporium spp. were present.     

Molecular Characterization of Diazotrophic and Denitrifying Bacteria Associated with Mangrove Roots

An analysis of the molecular diversity of N₂ fixers and denitrifiers associated with mangrove roots was performed using terminal restriction length polymorphism (T-RFLP) of nifH (N₂ fixation) and nirS and nirK (denitrification), and the compositions and structures of these communities among three sites were compared. The number of operational taxonomic units (OTU) for nifH was higher than that for nirK or nirS at all three sites. Site 3, which had the highest organic matter and sand content in the rhizosphere sediment, as well as the lowest pore water oxygen concentration, had the highest nifH diversity. Principal component analysis of biogeochemical parameters identified soil texture, organic matter content, pore water oxygen concentration, and salinity as the main variables that differentiated the sites. Nonmetric multidimensional scaling (MDS) analyses of the T-RFLP data using the Bray-Curtis coefficient, group analyses, and pairwise comparisons between the sites clearly separated the OTU of site 3 from those of sites 1 and 2. For nirS, there were statistically significant differences in the composition of OTU among the sites, but the variability was less than for nifH. OTU defined on the basis of nirK were highly similar, and the three sites were not clearly separated on the basis of these sequences. The phylogenetic trees of nifH, nirK, and nirS showed that most of the cloned sequences were more similar to sequences from the rhizosphere isolates than to those from known strains or from other environments.     

Nickel Tolerance and Accumulation by Filamentous Fungi From Sludge of Metal Finishing Industry

Industrialization, urbanization and increased vehicular traffic have resulted in increased contamination of our environment by heavy metals. The long persistence of heavy metals in nature has in turn resulted in development of metal resistant microbial strains. These strains are minimizing heavy metals toxicity, either by metal complexation or precipitation and other mechanisms. Characterization of fungal diversity was done in contaminated soil of the Wazirpur industrial area throughout the year. In this area highly acidic hazardous solid waste produced high concentration of heavy metals (Ni, Cu, Cr, Fe, Mn). Nickel toxicity is a major environmental concern. Due to long persistence of this waste in the environment without any treatment, many fungal isolates from the surrounding environment settle on the upper surface of waste. Few of them are capable of growing in the toxic conditions. More than 20 strains were isolated, most of them belonging to species of Aspergillus, Penicillium, Fusarium and Mucor genera. Seasonal variation in fungal diversity was significant. Four filamentous fungal isolates were found to be resistant for nickel (II) and a strain of Papulaspora sepedonoides reported first time for bioremediation of Ni (II) in this investigation, which is absorbing 62.33 μmol Ni gr^?1. These fungal isolates showed a high level (100–10000 mg kg^?1) of resistance for Ni (II) salt and removing Ni (II) from solution. Metal uptake varied with fungi. The toxicity also was influenced by different factors like pH and composition of growth medium.     

葡萄生单轴霉菌围可培养微生物多样性及潜在生防菌研究

【目的】揭示葡萄生单轴霉(Plasmoparaviticola)菌围可培养细菌和真菌的多样性特征,筛选对葡萄霜霉病有较强稳定防治效果的生防菌。【方法】连续两年从我国南北方具有代表性的7个葡萄产区采集葡萄霜霉病叶,镊子夹取经保湿培养获得的新鲜霉层并配制孢子囊悬浮液,采用传统分离培养法,结合形态分类、BOX-PCR指纹图谱分析以及分子鉴定结果,对葡萄生单轴霉菌围的可培养细菌和真菌进行聚类分析;采用菌株及其发酵液与病原菌孢子囊悬浮液等体积混合培养测定其对孢子囊的抑制作用,离体叶片接种法检测该菌株及其发酵液对霜霉病的防治效果。【结果】分离获得了90株细菌和110株真菌,分别归属于8个细菌属和14个真菌属,且相同地区不同葡萄品种葡萄生单轴霉菌围的细菌和真菌在同年处于同一分支。假单胞菌属(Pseudomonas)和枝孢属(Cladosporium)稳定存在于各地区不同品种葡萄霜霉病叶上葡萄生单轴霉菌围;在两年间稳定存在的菌株占比多数在80.0%以上且均具有较高的生防作用;其中,广泛分布的6株枝顶孢属(Acremonium)真菌对葡萄霜霉病的防治效果均较好,最高可达100.0%;防治效果较高的11个菌株的无菌发酵液中,黑曲霉(Aspergillusniger) NX2F、苋楔孢黑粉菌(Thecaphora amaranthi) BJ1G和匍枝根霉(Rhizopus stolonifer) BM1L的无菌发酵液防治效果均为100.0%。【结论】葡萄生单轴霉菌围的可培养细菌和真菌群落主要受地区因素影响,有较高的稳定性和生防作用,揭示了枝顶孢属真菌在我国葡萄主要产区葡萄生单轴霉菌围附生的普遍性,为葡萄霜霉病的防治提供了丰富和宝贵的资源。     

Diversity and spatial–temporal distribution of airborne fungi at the world culture heritage site Maijishan Grottoes in China

The deposition of the airborne microorganisms onto cultural heritage is associated closely with the subsequent biodeterioration. In this study, a systematic investigation was carried out to detect the seasonal variation and diversity of airborne fungal concentration at the World Cultural Heritage Site Maijishan Grottoes in western China. A bio-aerosol sampler was deployed to collect samples over four seasons in 2016. The culturable airborne fungi were isolated, purified and then identified with the extraction of genomic DNA, PCR amplification of ITS rRNA region, sequencing, and phylogenetic analysis. The concentrations of culturable fungi ranged from 216 to 1389 CFU/m³, which varied seasonally with significant differences among the sampling sites. Fifteen different fungal genera were confirmed, among them, Cladosporium was the most predominant fungal genus, followed by Penicillium. The fungal community structure and their relationship with environmental factors were also delineated. The spatial–temporal differences of airborne fungi at Maijishan Grottoes were mainly due to height, rainfall, relative humidity, and temperature. The dominant genera Cladosporium and Penicillium may pose potential threats to the ancient painted sculptures and murals, and monitoring of the airborne fungi at such a heritage site could provide supporting data for the pre-warning and control of fungal outbreaks inside the caves for better management.