Analysis Of Fungal Diversity Research Articles

Analysis of fungal diversity in the feces of Arborophila rufipectus.

Intestinal fungal composition plays a crucial role in modulating host health, and thus is of great significance in the conservation of endangered bird species. However, research on gut fungal composition in birds is limited. Therefore, in this study, we aimed to examine gut fungal community and potential fecal pathogen composition in wild Arborophila rufipectus. Fecal samples were collected from the habitats of wild A. rufipectus and Lophura nycthemera (a widely distributed species belonging to the same family as A. rufipectus) in summer and autumn. Thereafter, RNA was collected and the internal transcribed spacer rRNA gene was sequenced via high-throughput sequencing to investigate seasonal variations in intestinal core fungi, microbial fungi, and potential pathogenic fungi. The gut microbiota of A. rufipectus and L. nycthemera were highly similar and mainly consisted of three phyla, Ascomycota (58.46%), Basidiomycota (28.80%), and Zygomycota (3.56%), which accounted for 90.82% of the fungal community in all the samples. Further, the predominant genera were Ascomycota_unclassified (12.24%), Fungi_unclassified (8.37%), Davidiella (5.18%), Helotiales_unclassified (2.76%), Wickerhamomyces (1.84%), and Pleosporales_unclassified (1.14%), and the potential fecal pathogens identified included Candida, Cryptococcus, Trichosporon, and Malassezia. Our results provide evidence that the diversity of intestinal fungi in the endangered species, A. rufipectus, is similar to that in the common species, L. nycthemera, and may serve as a basis for monitoring the status of A. rufipectus and for developing conservation measures.

The diversity of fungal associates of Dendrobium ovatum (L.) Kraenzl., an endemic orchid of the Western Ghats of India

Dendrobium ovatum is a tropical epiphytic orchid endemic to the Western Ghats of India and has been listed as a threatened species in recent research due to its declining populations and changes in flowering and fruit set patterns. This study aims to investigate the mycoflora associated with the roots, stems and leaves of D. ovatum. Both surface‐associated and endophytic fungal associates were isolated and identified using morphological and molecular methods. The study resulted in the isolation of 139 cultures, which were divided into 24 morphotypes, 99% of which belonged to Ascomycota. The most dominant members, Trichoderma harzianum and Colletotrichum gloeosporioides, were consistently observed across all the study sites. Tissue‐specific fungal diversity analysis revealed that each organ was dominated by a distinct fungal group, forming characteristic communities specific to each tissue. The roots of D. ovatum exhibited the highest species richness and diversity, compared to the stem and leaves. This research also represents the first documentation of fungal associates of the threatened orchid D. ovatum.

Analysis of Fungal Diversity, Physicochemical Properties and Volatile Organic Compounds of Strong-Flavor Daqu from Seven Different Areas.

Strong-flavor Daqu, as a fermentation agent, plays a significant role in shaping the quality of strong-flavor baijius, and fungal species in Daqu are important factors affecting the quality of Daqu. Therefore, we selected strong-flavor Daqu from seven different origins to study the fungal composition and the effects of the fungal composition on the physicochemical properties and volatile organic compounds (VOCs). It was found that the fungal composition influences the physicochemical properties of Daqu. Specifically, there was a positive link between Rhizomucor, Rhizopus, Thermomyces, and liquefying activity and a positive correlation between Aspergillus and fermenting activity. Furthermore, the relationships between esterifying activity and Thermomyces, Rhizomucor, Aspergillus, Pichia, and Saccharomycopsis were found to be positive. The VOCs in Daqu were affected by Aspergillus, Issatchenkia, Pichia, and Thermoascus. Issatchenkia was significantly positively correlated with benzeneethanol as well as Aspergillus and pentadecanoic acid ethyl ester, ethyl myristate. Pichia and Thermoascus were significantly negatively correlated with benzaldehyde and 2-furaldehyde. This study deepens our understanding of the relationship between VOCs, the physicochemical properties with microbial communities, and reference significance for the production of better-quality strong-flavor Daqu.

Analysis of fungal diversity in the gut feces of wild takin (Budorcas taxicolor).

The composition of the intestinal microbiome correlates significantly with an animal's health status. Hence, this indicator is highly important and sensitive for protecting endangered animals. However, data regarding the fungal diversity of the wild Budorcas taxicolor (takin) gut remain scarce. Therefore, this study analyzes the fungal diversity, community structure, and pathogen composition in the feces of wild B. taxicolor. To ensure comprehensive data analyses, we collected 82 fecal samples from five geographical sites. Amplicon sequencing of the internal transcribed spacer (ITS) rRNA was used to assess fecal core microbiota and potential pathogens to determine whether the microflora composition is related to geographical location or diet. We further validated the ITS rRNA sequencing results via amplicon metagenomic sequencing and culturing of fecal fungi. The fungal diversity in the feces of wild Budorcas taxicolor primarily comprised three phyla (99.69%): Ascomycota (82.19%), Fungi_unclassified (10.37%), and Basidiomycota (7.13%). At the genus level, the predominant fungi included Thelebolus (30.93%), Functional_unclassified (15.35%), and Ascomycota_unclassified (10.37%). Within these genera, certain strains exhibit pathogenic properties, such as Thelebolus, Cryptococcus, Trichosporon, Candida, Zopfiella, and Podospora. Collectively, this study offers valuable information for evaluating the health status of B. taxicolor and formulating protective strategies.

Analysis of fungal diversity and structure in Nile River water polluted with crude oil and naphthalene using microcosm experiments

Analysis of fungal diversity and structure in Nile River water polluted with crude oil and naphthalene using microcosm experiments

Comparative analysis of gut fungal composition and structure of the yaks under different feeding models.

The yaks that inhabit the Tibetan plateau are a rare breed that is closely related to local economic development and human civilization. This ancient breed may have evolved a unique gut microbiota due to the hypoxic high-altitude environment. The gut microbiota is susceptible to external factors, but research regarding the effects of different feeding models on the gut fungal community in yaks remains scarce. In this study, we compared and analyzed the composition and variability of the gut fungal community among wild yaks (WYG), house-feeding domestic yaks (HFG), and grazing domestic yaks (GYG). The results revealed that Basidiomycota and Ascomycota were the most preponderant phyla in the gut fungal community, regardless of feeding models. Although the types of dominant fungal phyla did not change, their abundances did. Intergroup analysis of fungal diversity showed that the Shannon and Simpson indices of WYG and GYG were significantly higher than those of HFG. Fungal taxonomic analysis showed that there were 20 genera (Sclerostagonospora and Didymella) that were significantly different between WYG and GYG, and 16 genera (Thelebolus and Cystobasidium) that were significantly different between the WYG and HFG. Furthermore, the proportions of 14 genera (Claussenomyces and Papiliotrema) significantly decreased, whereas the proportions of eight genera (Stropharia and Lichtheimia) significantly increased in HFG as compared to GYG. Taken together, this study indicated that the gut fungal composition and structure differ significantly between yaks raised in different breeding groups.

Addition of Organic Matter to Pine Plantations on Agricultural Land Positively Alters the Mycobiome of Agricultural Soils

Afforestation of former agricultural land poses a real challenge for foresters because soil life is often severely limited by the loss of natural soil fungal diversity. In addition, former agricultural soils have low levels of fungal species typical of forest soils, which have a unique microbiome that plays a protective role (antagonists, plant growth promoters, mycorrhizal fungi, etc.). This study aimed to determine the effect of using organic material in the form of bark compost, wood waste, and sawdust to improve the soil mycobiome of soils that have been damaged by their agricultural use. This study used experimental plots established 20 years ago, and we compared the biodiversity of the treated soils with that of the control soils by analysing soil samples with powerful molecular methods. Next-generation sequencing analysis of DNA extracted from soil samples and subsequent analysis of their species composition and biodiversity showed that the mycobiome of soil fungi has been altered by the addition of various forms of organic material. The proportion of fungi belonging to the Ascomycota decreased in favour of species from the Basidiomycota and Mucoromycota. The dominant fungal groups in the soil of the control area were Sagenomella, Wilcoxina, Oidiodendron, Meliniomyces, and Penicillium. Enrichment with organic matter by adding bark compost under the roots led to an increase in Penicillium, Inocybe, and Amphinema. The application of bark compost on the surface led to an increase in the dominance of Inocybe fungi in the soil. The mycobiome of the plant to which woody debris was applied was characterised by a marked dominance of fungi of the genera Russula, Oidiodendron, and Penicillium. Similar ratios were found in the plant to which sawdust was applied, where the fungi Meliniomyces, Penicillium, Oidiodendron, and Russula dominated. A comparative analysis of fungal diversity with the Shannon diversity index showed that the most diverse fungal communities were found in the sawdust plant (6.56), while the control sample (a soil sample from an agricultural area where no organic material was applied) had an index of 5.71. After the treatments, more potential antagonists against pine pathogens and mycorrhizal fungi were found to form beneficial symbiotic relationships with them. In our opinion, the results of this study show that it is worthwhile to introduce different forms of organic matter to post-agricultural land to improve soil biodiversity and mycorrhizal associations of pine roots with fungi to ensure the sustainability of the first generation of forests created.

Fungal Diversity and Community Composition across Ecosystems

Fungi have shaped the biosphere since the development of life on Earth. Despite fungi being present in all environments, most of the available fungal research has focused on soils. As a result, the role and composition of fungal communities in aquatic (marine and freshwater) environments remain largely unexplored. The use of different primers to characterise fungal communities has additionally complicated intercomparisons among studies. Consequently, we lack a basic global assessment of fungal diversity across major ecosystems. Here, we took advantage of a recently published 18S rRNA dataset comprising samples from major ecosystems (terrestrial, freshwater, and marine) to attempt a global assessment of fungal diversity and community composition. We found the highest fungal diversities for terrestrial > freshwater > marine environments, and pronounced gradients of fungal diversity along temperature, salinity, and latitude in all ecosystems. We also identified the most abundant taxa in each of these ecosystems, mostly dominated by Ascomycota and Basidiomycota, except in freshwater rivers where Chytridiomycota dominated. Collectively, our analysis provides a global analysis of fungal diversity across all major environmental ecosystems, highlighting the most distinct order and ASVs (amplicon sequencing variants) by ecosystem, and thus filling a critical gap in the study of the Earth's mycobiome.

Analysis of Fungal Diversity of Three Samples (dqjq_ck, dqjqcp and dqjp3) from Danquan Distillery

There are fungal communities in fermented grains and high-temperature Daqu of Danquan Baijiu, which had an important effect on Baijiu production. As a saccharifying and fermenting material, to a great extent, Daqu determines Baijiu flavour. However, in high-temperature Daqu “dqjq_ck” and “dqjqcp” (after fermentation for 45 d, 135 d) and fermented grains dqjp3 originated from Danquan distillery, fungal communities remain unclear. Therefore, this study analyzed fungal communities by high-throughput sequencing. Their fungal communities mainly included Ascomycota at phylum level. The most abundant fungal genera were Aspergillus, Issatchenkia and Zygosaccharomyces in dqjq_ck, dqjqcp, dqjp3, respectively. The dominant genera included Aspergillus (96.21%), Thermomyces (2.13%) in dqjq_ck. The dominant genera contained Issatchenkia (48.29%), Millerozyma (14.53%), Thermoascus (12.17%), Aspergillus (8.80%), Hyphopichia (3.28%), Rhizomucor (3.17%), Lichtheimia (2.62%), Thermomyces (2.27%), Monascus (1.37%) in dqjqcp. The dominant genera had Zygosaccharomyces (68.48%), Monascus (15.18%), Aspergillus (6.97%), Cladosporium (1.12%) in dqjp3. The main fungi included Aspergillus, Zygosaccharomyces, Issatchenkia, Monascus, Millerozyma, Thermoascus, Thermomyces, Hyphopichia, Rhizomucor, Lichtheimia and Cladosporium in three samples. The fungal succession of three samples were revealed in three key stages of Danquan Baijiu production. This paper will lay the basis for screening outstanding fungi to enhance Danquan Baijiu quality in near future.

Analysis of Fungal Diversity before and after Discoloration of Rubberwood in Xishuangbanna

The discoloration problem of rubber wood caused by the discoloration fungi has caused the degradation of rubber wood and greatly reduced its economic value, and the prevention and control of rubber wood discoloration have become the top priority of basic research on rubber wood protection and modification. To determine the fungal community diversity and dominant groups before and after discoloration of rubber wood, nine rubber wood samples were subjected to ITS sequencing using Illumina high-throughput sequencing technology. The results showed that the detected fungal communities comprised 5 phyla, 18 classes, 58 orders, 137 families, 218 genera, and 297 species. Discoloration of rubber wood is not caused by a single species, with the dominant genera for discolored rubber wood being Huntiella, Ceratocystis, and Acremonium and for undiscolored rubber wood, Phomopsis. Furthermore, the diversity, uniformity of species distribution, and richness of discolored rubber wood were found to be higher than those of undiscolored rubber wood. In conclusion, understanding the change trends in the structure of these fungal communities is essential for studying the biological control of rubberwood discoloration.

Antarctic Glacial Meltwater Impacts the Diversity of Fungal Parasites Associated With Benthic Diatoms in Shallow Coastal Zones.

Aquatic ecosystems are frequently overlooked as fungal habitats, although there is increasing evidence that their diversity and ecological importance are greater than previously considered. Aquatic fungi are critical and abundant components of nutrient cycling and food web dynamics, e.g., exerting top-down control on phytoplankton communities and forming symbioses with many marine microorganisms. However, their relevance for microphytobenthic communities is almost unexplored. In the light of global warming, polar regions face extreme changes in abiotic factors with a severe impact on biodiversity and ecosystem functioning. Therefore, this study aimed to describe, for the first time, fungal diversity in Antarctic benthic habitats along the salinity gradient and to determine the co-occurrence of fungal parasites with their algal hosts, which were dominated by benthic diatoms. Our results reveal that Ascomycota and Chytridiomycota are the most abundant fungal taxa in these habitats. We show that also in Antarctic waters, salinity has a major impact on shaping not just fungal but rather the whole eukaryotic community composition, with a diversity of aquatic fungi increasing as salinity decreases. Moreover, we determined correlations between putative fungal parasites and potential benthic diatom hosts, highlighting the need for further systematic analysis of fungal diversity along with studies on taxonomy and ecological roles of Chytridiomycota.

Multiomics analysis reveals that peach gum colouring reflects plant defense responses against pathogenic fungi

In the present study, the differences in the antioxidant capability, metabolite composition and fungal diversity in peach gum with various colours were investigated. Metabolomics revealed that peach gum comprised many small-molecule metabolites (including primary and secondary metabolites), and most polyphenols (such as flavonoids and phenolic acids) showed a significantly positive relationship with the colour deepening, total phenol content and antioxidant capability. Using fungal diversity analysis, the abundance of five fungi at the genus level increased with peach gum colour deepening, and these fungi demonstrated a significantly positive relationship with two defense hormones (salicylic acid and abscisic acid) and most polyphenols (particularly flavonoids). The gummosis pathogenic fungus Botryosphaeria was among the five fungi, suggesting that peach gum colouring may reflect plant defense responses against pathogenic fungi. Additionally, the concentrations of 12 flavonoids in peach gum samples were detected based on LC-QQQ/MS, among which hesperetin, naringenin and eriodictyol were the most abundant.

Crowdsourcing fungal biodiversity: revision of iNaturalist observations in Northwestern Siberia

The paper presents the first analysis of crowdsourcing data of all observations of fungi (including lichens) and myxomycetes in Northwestern Siberia uploaded to iNaturalist.org to date (24.02.2022). The Introduction presents an analysis of fungal diversity crowdsourcing globally, in Russia, and in the region of interest. Materials and methods describe the protocol of uploading data to iNaturalist.org, the structure of the crowdsourcing community, initiative to revise the accumulated data, procedures of data analysis, and compilation of a dataset of revised crowdsourced data. The Results present the analysis of accumulated data by several parameters: temporal, geographical and taxonomical scope, observation and identification efforts, identifiability of various taxa, species novelty and Red Data Book categories and the protection status of registered observations. The Discussion provides data on usability of crowdsourcing data for biodiversity research and conservation of fungi, including pros and contras. The Electronic Supplements to the paper include an annotated checklist of observations of protected species with information on Red Data Book categories and the protection status, and an annotated checklist of regional records of new taxa. The paper is supplemented with a dataset of about 15 000 revised and annotated records available through Global Biodiversity Information Facility (GBIF). The tradition of crowdsourcing is rooted in mycological societies around the world, including Russia. In Northwestern Siberia, a regional mycological club was established in 2018, encouraging its members to contribute observations of fungi on iNaturalist.org. A total of about 15 000 observations of fungi and myxomycetes were uploaded so far, by about 200 observers, from three administrative regions (Yamalo-Nenetsky Autonomous Okrug, Khanty-Mansi Autonomous Okrug, and Tyumen Region). The geographical coverage of crowdsourcing observations remains low. However, the observation activity has increased in the last four years. The goal of this study consisted of a collaborative effort of professional mycologists invited to help with the identification of these observations and analysis of the accumulated data. As a result, all observations were reviewed by at least one expert. About half of all the observations have been identified reliably to the species level and received Research Grade status. Of those, 90 species (195 records) represented records of taxa new to their respective regions; 876 records of 53 species of protected species provide important data for conservation programmes. The other half of the observations consists of records still under-identified for various reasons: poor quality photographs, complex taxa (impossible to identify without microscopic or molecular study), or lack of experts in a particular taxonomic group. The Discussion section summarises the pros and cons of the use of crowdsourcing for the study and conservation of regional fungal diversity, and summarises the dispute on this subject among mycologists. Further research initiatives involving crowdsourcing data must focus on an increase in the quality of observations and strive to introduce the habit of collecting voucher specimens among the community of amateurs. The timely feedback from experts is also important to provide quality and the increase of personal involvement.

Integrated Bacterial and Fungal Diversity Analysis Reveals the Gut Microbial Alterations in Diarrheic Giraffes.

Gut microbiota has been demonstrated to be associated with multiple gastrointestinal diseases, but information regarding the gut microbial alternations in diarrheic giraffe remains scarce. Here, 16S rDNA and ITS gene amplicon sequencing were conducted to investigate the gut microbial composition and variability in diarrheic giraffes. Results demonstrated that Firmicutes and Proteobacteria were the most dominant phyla in the gut bacterial community, whereas Ascomycota and Basidiomycota were observed to be predominant in the gut fungal community regardless of health status. However, the species and relative abundance of preponderant bacterial and fungal genera in healthy and diarrheic giraffes were different. In contrast to the relatively stabilized gut fungal community, gut bacterial community displayed a significant decrease in the alpha diversity, accompanied by distinct changes in taxonomic compositions. Bacterial taxonomic analysis revealed that the relative abundances of eight phyla and 12 genera obviously increased, whereas the relative abundances of two phyla and eight genera dramatically decreased during diarrhea. Moreover, the relative richness of five fungal genera significantly increased, whereas the relative richness of seven fungal genera significantly declined in diarrheic giraffes. Taken together, this study demonstrated that diarrhea could cause significant alternations in the gut microbial composition of giraffes, and the changes in the gut bacterial community were more significant than those in the gut fungal community. Additionally, investigating the gut microbial characteristics of giraffes in different health states is beneficial to provide a theoretical basis for establishing a prevention and treatment system for diarrhea from the gut microbial perspective.

Microbiological analysis of water and fish of fish farms in the protection area of the western lowlands of Maranhão, Brazil

This work aimed to analyse the water quality and identify the fungal diversity present in the skin and gills of tambatinga fish from the environmental protection area of the western lowlands of Maranhão, Brazil. The physicochemical parameters of water were analysed and microbiological tests were carried out to identify the most probable number of total coliforms and Escherichia coli. Forty tambatinga specimens were collected. In addition, skin and gill fragments were removed for fungi isolation in culture medium (BDA). Physicochemical and microbiological parameters of water, fish biometry and fungal diversity of fish in the dry and rainy periods were analysed. Results showed significant differences in the total length (TC) and weight (P) of fish in the dry and rainy periods. Microbiological data showed that the water of the four fish farms analysed in both periods are within the limits established by the National Environment Council (CONAMA) of Brazil and within limits established in complementary literature. Regarding the physicochemical parameters of water, only conductivity (F (1, 14) = 8.611; p = 0.011) and temperature had considerable differences (F (1, 14) = 11.037; p = 0.005). There was prevalence of yeasts (33%) and fungi of the genus Fusarium sp. (26%), Penicillium sp. (21%) and Aspegillus sp. (20%). The analysis of fungal diversity showed associations for sex, Aspegillus sp. (X2 = 19,600; DF = 1; p = 0,001) and the dry season, Penicillium sp. (X2 = 8,120; DF = 1; p = 0,004) and Fusarium sp. (X2 = 21.583; GL = 1; p = 0.001). It is concluded, therefore, that the harvests are within the established by the legislation of the National Environment Council—CONAMA in Brazil and complementary literature. However, it is necessary to evaluate the presence of possible mycotoxins in cultivated fish, since the most prevalent fungal species are considered to be mycotoxigenic fungi and are widely disseminated in the environment.

Comparative Analysis of Fungal Diversity and Community Structure of Different Volcanic Soils in Wudalianchi, China

The relationship between the fungal community characteristics and soil environmental factors of volcanic ecosystem in Wudalianchi, China were investigated. The soil fungal community structure and diversity of new, old, and non-erupting volcanos were explored through highthroughput sequencing technology. The result showed that the physical and chemical properties of three plots were significantly different. Through sequencing 578 species, 366 genera, 202 families, 89 orders, 32 classes, and 11 phyla were detected. Among them Ascomycota and Basidiomycota were the dominant fungi phyla. The relative abundance of various flora determined by phylum classification showed significant differences. The Shannon, Simpson, Ace, and Chao1 indices for the soil fungi in the three plots were also significantly different. Redundancy and correlation analyses showed that the α diversity of fungi was significantly correlated with pH, organic matter and total nitrogen in the soil. These results indicate that soil environmental factors influence the fungal diversity in the different volcanic ecosystems in Wudalianchi, China.
 Bangladesh J. Bot. 50(2): 327-334, 2021 (June)

High-throughput sequencing-based analysis of fungal diversity and taste quality evaluation of Douchi, a traditional fermented food.

Douchi, a popular traditional fermented soybean product, is mainly made by natural fermentation. However, its taste quality is affected by specific fungal communities which vary greatly according to fermentation conditions and production technologies used in different regions. Therefore, the taste quality of Douchi samples from different regions was digitally evaluated using electronic tongue technology. In addition, the fungal community structures and its association of them were also identified using high‐throughput sequencing technology. Results showed that there were obvious differences in the taste quality of samples from different regions, while the tastes of different types of samples from the same region were similar. Sourness, umami, richness, and saltiness were the main reasons for regional differences in taste. Similarly, the results of high‐throughput sequencing indicated that samples from different regions displayed important differences in dominant fungal genus, among which Debaryomyces, Fusarium, Pichia, Aspergillus, and Saccharomyces were the main dominant fungi. Debaryomyces and Trichosporon were conducive to the formation of taste qualities of Douchi, while Cladosporium and Candida have a negative impact on the taste quality of Douchi var correlation analysis. This study indicated the effects of dominant fungi on the formation of Douchi taste quality, allowing a deeper understanding of the role of microbial species in generating fermented soybean products in China. Our work provides theoretical support to guide the improvement of the industrial production process of Douchi.

Fungal Diversity Analysis of Grape Musts from Central Valley-Chile and Characterization of Potential New Starter Cultures.

Autochthonous microorganisms are an important source of the distinctive metabolites that influence the chemical profile of wine. However, little is known about the diversity of fungal communities associated with grape musts, even though they are the source of local yeast strains with potential capacities to become starters during fermentation. By using internal transcribed spacer (ITS) amplicon sequencing, we identified the taxonomic structure of the yeast community in unfermented and fermented musts of a typical Vitis vinifera L. var. Sauvignon blanc from the Central Valley of Chile throughout two consecutive seasons of production. Unsurprisingly, Saccharomyces represented the most abundant fungal genus in unfermented and fermented musts, mainly due to the contribution of S. uvarum (42.7%) and S. cerevisiae (80%). Unfermented musts were highly variable between seasons and showed higher values of fungal diversity than fermented musts. Since microbial physiological characterization is primarily achieved in culture, we isolated nine species belonging to six genera of fungi from the unfermented must samples. All isolates were characterized for their potential capacities to be used as new starters in wine. Remarkably, only Metschnikowia pulcherrima could co-exist with a commercial Saccharomyces cerevisiae strain under fermentative conditions, representing a feasible candidate strain for wine production.

Comparative Analysis of Fungal Diversity in Rhizospheric Soil from Wild and Reintroduced Magnolia sinica Estimated via High-Throughput Sequencing

Magnolia sinica is a critically endangered species and considered a “plant species with extremely small populations” (PSESP). It is an endemic species in southeastern Yunnan Province, China, with reproductive barriers. Rhizosphere fungi play a crucial role in plant growth and health. However, the composition, diversity, and function of fungal communities in wild and reintroduced M. sinica rhizospheres remain unknown. In this study, Illumina sequencing of the internal transcribed spacer 2 (ITS2) region was used to analyze rhizospheric soil samples from wild and reintroduced M. sinica. Thirteen phyla, 45 classes, 105 orders, 232 families, and 433 genera of fungi were detected. Basidiomycota and Ascomycota were dominant across all samples. The fungal community composition was similar between the wild and reintroduced rhizospheres, but the fungal taxa relative abundances differed. The fungal community richness was higher in the reintroduced rhizosphere than in the wild rhizosphere, but the diversity showed the opposite pattern. Soil nutrients and leaf litter significantly affected the fungal community composition and functional diversity. Here, the composition, structure, diversity, and ecological functions of the fungal communities in the rhizospheres of wild and reintroduced M. sinica were elucidated for the first time, laying a foundation for future research and endangered species protection.

COMPARISON EFFICACY OF ITS AND 18S rDNA PRIMERS FOR DETECTION OF FUNGAL DIVERSITY IN COMPOST MATERIAL BY PCR-DGGE TECHNIQUE

Through composting process, biosolid wastes are gradually transformed into compost material which can be used as soil fertilizer. Among microorganisms involved in composting process, fungi play important roles because they break down complex substrates, such as ligno-cellulose. Recently, PCR-DGGE technique has been considered as a useful tool for analysis of fungal diversity in environmental samples. Among other factors, primer set selection is necessary for successful of the PCR-DGGE analysis. There are several PCR primer sets targeting fungal variable regions of 18S ribosomal DNA (rDNA) and internal transcribed spacer (ITS) for the use in community analyses, however there exist just few reports on efficacy of these primers in studying fungal communities in compost materials. In this study, four different primer sets were tested, including EF4/Fung5 (followed by EF4/NS2-GC), EF4/ITS4 (followed by ITS1F-GC/ITS2), NS1/GC-Fung, and FF390/FR1-GC. Extracted DNA from compost materials often contains co-extracted humic substances and other PCR inhibitors. Therefore, the primers were tested for (i) tolerance to the PCR inhibitors presenting in the DNA extracted from compost materials, and (ii) efficacy and specificity of the PCR. The results showed that of the four primer sets, only FF390/FR1-GC achieved both criteria tested whereas the other three did not, i.e. primer EF4/ITS4 had low tolerance to PCR inhibitors, primers EF4/Fung5 was low in PCR amplification efficacy, whereas primers EF4/ITS4 created unspecific products. DGGE analyses of PCR products amplified with the primer set FF390/FR1-GC showed single bands for reference pure cultures Penicillium sp., Aspergillus sp., and Trichoderma sp., as well as distinctly separated bands for the fungal communities of three different composting materials. Thus, the primer set FF390/FR1-GC could be suitable for studying structure and dynamic of fungal communities in compost materials.