云南松林野生食用菌物种多样性及保护对策

近年来,随着野生食用菌的商业化采收规模急剧扩大,云南松（Pinus yunnanensis）林下野生食用菌尤其是野生贸易真菌的种类和产量呈逐年下降趋势。作者于2000－2004年夏,对滇中及其邻近地区云南松林下野生食用菌的资源状况进行了调查研究。作者共采集、鉴定标本457号,并对他人采自云南松林下的152号馆藏标本也进行了重新鉴定,共统计野生食用菌27科39属147种（含变种、变型）;同时对市场上出售的采自云南松林下的野生食用菌类群也进行了调查统计,共收集、鉴定标本167号,计21科33属97种（含变种、变型）。调查结果显示,目前记录云南松林产野生食用菌共计27科43属211种（含变种、变型）,野生贸易真菌有23科35属111种（含变种、变型）。其中红菇科（Russulaceae）的种数最多,包括红菇属（Russula）32种,乳菇属（Lactarius）17种,分别占总种数的15.17%和8.06%;牛肝菌科（Boletaceae）占第二位,有41种,占总种数的19.43%。其次是口蘑科（Tricholomataceae）17种,占总种数的8.06%。丛枝瑚菌科（Ramariaceae）有11种,乳牛肝菌科（Suillaceae）11种,鸡油菌科（Cantharellaceae）9种,三者的种数占总种数的百分比分别为5.21%、5.21%和4.27%。在这些类群当中,红菇属、牛肝菌属（Boletus）、乳菇属、乳牛肝菌属（Suillus）、枝瑚菌属（Ramaria）、口蘑属（Tricholoma）、鸡油菌属（Cantharellus）和革菌属（Thelephora）的30余种真菌为云南市场上最为常见的野生贸易类群,约占云南优势野生贸易真菌种数的47.50%。作者呼吁立即采取行之有效的措施保护和管理这些珍贵的自然资源,并加强研究,以期达到持续利用之目的。     

云南普洱地区野生食药用菌及其持续利用

2002—2009年夏,对云南省普洱市澜沧县、景东县及其周边地区野生食用菌的物种多样性、用途或特性以及濒危等级进行了调查研究,共采集、鉴定标本593份,并对采于该区的214份馆藏标本进行了统计,野生食用菌共计35科61属134种(含变种、变型)。同时对该区市场上出售的野生贸易类群也进行了调查统计,有28科39属95种(含变种、变型)。迄今普洱共计有野生食用菌38科67属155种(含变种、变型),其中多孔菌科(Polyporaceae)所含种数最多,有17种,占总种数的10.97%;红菇科(Russulaceae)和牛肝菌科(Bo-letaceae)皆为16种,共占总种数的20.65%,灵芝菌科(Ganodermataceae)有11种,占总种数7.1%。在这些类群当中,木耳属(Auricularia)、牛肝菌属(Boletus)、乳菇属(Lactarius)、枝瑚菌属(Ramaria)、红菇属(Russula)、蚁巢菌属(Termitomyces)和革菌属(Thelephora)等属的42种真菌为普洱市场上最常见的野生贸易类群,约占云南优势野生贸易真菌总种数的52.5%。近年来,随着人们对野生食用菌消费需求的日益增长,导致其商业化采收规模急剧扩大,造成生存环境的破坏甚至丧失,从而使普洱野生食用菌,尤其是野生贸易类群的产量呈逐年下降趋势。因此,亟待采取行之有效的措施保护和管理这些珍贵的自然资源,以期达到持续利用之目的。     

Natural occurrence in Argentina of a new fungal pathogen of cockroaches,Metarhizium argentinense sp. nov.

The aim of this study was to search for entomopathogenic fungi that infect wild cockroaches in forest ecosystems in two protected natural areas of Argentina. Two isolates of Metarhizium argentinense were obtained and identified from wild cockroaches (Blaberidae: Epilamprinae) through the use of morphological characteristics and molecular phylogenetic analyses. This novel species was found in Argentina and is a member of the Metarhizium flavoviride species complex. Phylogenetic analyses, based on sequence similarity analysis using internal transcribed spacer (ITS) and a set of four protein-coding marker sequences (EF1A, RPB1, RPB2 and BTUB), supported the status of this fungus as a new species. In addition, we tested the biological activity of the new species through assays against Blattella germanica nymphs and found that the two evaluated isolates were pathogenic. However, isolate CEP424 was more virulent and caused a confirmed mortality of 76 % with a median lethal time of 7.2 d. This study reports the southernmost worldwide location of a Metarhizium species that infects cockroaches and will help expand the knowledge of the biodiversity of pathogenic fungi of Argentine cockroaches.     

Multi-gene phylogenetic analyses reveals <Emphasis Type="Italic">Neohelicosporium</Emphasis> gen. nov. and five new species of helicosporous hyphomycetes from aquatic habitats

Helicosporous hyphomycetes are a morphologically allied group of Tubeufiales. We introduce a new helicosporous genus, Neohelicosporium, with five new species, Neohelicosporium aquaticum, N. guangxiense, N. hyalosporum, N. parvisporum, and N. thailandicum, based on morphological and phylogenetic evidence. The RPB2 protein gene data are provided to analyze their phylogeny in Tubeufiales. Phylogenetic analyses of combined ITS, LSU, RPB2, and TEF1α sequence data from 13 new isolates of Neohelicosporium provided evidence to support the establishment of the new taxa. The morphological characters of Neohelicosporium that differentiate it from other helicosporous species are compared and discussed.     

In vitro mycorrhization and acclimatization of Amanita caesareoides and its relatives on Pinus densiflora

Amanita caesareoides is a sister species of Amanita caesarea, also known as Caesar’s mushroom and one of the most desirable edible mycorrhizal mushrooms. However, cultivation of Caesar’s mushrooms has not yet been successful due to the difficulties involved in establishing pure cultures. In this study, we established pure cultures of four Asian Caesar’s mushroom species, i.e., A. caesareoides, Amanita javanica, Amanita esculenta, and Amanita similis, which were identified by sequence analysis of their rDNA internal transcribed spacer (ITS) region. Five selected isolates in A. caesareoides, A. javanica, and A. esculenta were tested for ectomycorrhizal syntheses with axenic Pinus densiflora seedlings in vitro. Ectomycorrhizal tips of each fungal isolate tested were observed on pine lateral roots within 5 months of inoculation. Seventeen pine seedlings that formed ectomycorrhizas in vitro with these three Amanita species were acclimatized under non-sterile conditions. Seven months following acclimatization, ectomycorrhizal colonization by A. caesareoides was observed on newly grown root tips, which was confirmed by polymerase chain reaction restriction fragment length polymorphism analysis of the fungal rDNA ITS region. Two other Amanita species also survived during ectomycorrhizal acclimatization. These results suggest that the cultivation of A. caesareoides and its relatives can be attempted through mycorrhizal synthesis using P. densiflora as a host. This is the first report of in vitro mycorrhization of Asian Caesar’s mushrooms and their acclimatization under non-sterile conditions.     

MSP-PCR and RAPD molecular biomarkers to characterize<Emphasis Type="Italic">Amanita ponderosa</Emphasis> mushrooms

Amanita ponderosa is a specie of wild edible mushrooms growing spontaneously in some Mediterranean microclimates, namely in Alentejo and Andaluzia, in the Iberian Peninsula. The nutritional values of these fungi make them highly exportable. Due to the wide diversity of mushrooms in nature, it is essential to differentiate and to identify the various edible species. RAPD markers have been used as a valuable tool to distinguish the different genotypes, although this method has not yet been used toAmanita ponderosa. Two methods were used to establish different genetic fingerprinting patterns of edible mushrooms. Samples ofAmanita ponderosa were collected in six different regions of the southwest of the Iberian Peninsula and compared by RAPD-PCR and MSP-PCR. Additionally, to compare molecular profiles with others genera of edible mushrooms, three species of Basidiomycetes (Pleurotus ostreatus, Lactarius deliciosus andCoriolus versicolor) and an Ascomycete were used. Results showed that some molecular markers discriminate among an Ascomycete from Basidiomycetes (Amanita ponderosa, Pleurotus ostreatus, Lactarius deliciosus andCoriolus versicolor) and discriminate among the different genera within basidiomycetes, as it is expected. Moreover, OPF-6, OPG-2, OPG3 and M13 primes allowed to unravel a level of genetic polymorphism withinAmanita ponderosa mushrooms collected from different geographic origin.     

Declining Wild Mushroom Recognition and Usage in Burkina Faso

Declining Wild Mushroom Recognition and Usage in Burkina Faso. Ethnomycological investigation was carried out in Burkina Faso, West Africa, using standardized interviews focused on 40 representative mushroom species assembled in a traveling herbarium (photographs plus preserved specimens). A total of 540 informants from 18 localities representing three main ethnic groups—Mòosé, Bobo, and Gouin—were interviewed, though the bulk of those interviewed were Mòosé (or Mossi) from the more populous central region of the country. Among the study species, as many as 24 were considered edible by the different ethnic groups; a few medicinal uses also were noted. The edibility of a common Chlorophyllum species is confirmed for the three ethnic groups. Mushroom nomenclature is not well developed in any of the three groups as evidenced by the fact that many mushrooms, including some common edible species, do not appear to have specific local names, but are instead simply referred to by a general term for “mushroom.” Preliminary observations about inter–ethnic variation in recognition, naming, and appreciation of mushrooms are noted. Older people recognized many more mushroom species than did younger people, and women appeared to be slightly more knowledgeable about mushrooms than men. The most important finding is that the gathering and consumption of wild edible mushrooms seems to be dying out in Burkina Faso, especially in the central part of the country, apparently because of declining mushroom populations resulting from disappearing forest habitats. NOTE TO VENDOR: In place of the corresponding author, send proof to this paper’s editor, David Aroro, maxfun @cruzio.com     

Molecular and morphological characterization of a poorly known marine ciliate, Myoschiston duplicatum precht 1935: implications for phylogenetic relationships between three morphologically similar genera -- Zoothamnium, Myoschiston, and Zoothamnopsis (Ciliophora, Peritrichia, Zoothamniidae)

We studied the morphology and molecular phylogeny of Myoschiston duplicatum, a peritrich ciliate that has been recorded as an epibiont of crustaceans, but which we also identified on marine algae from Korea. The important morphological characteristics revealed by silver staining of Myoschiston species have not been described because they are rarely collected. Using morphological methods, we redescribed the type species of the genus, Myoschiston duplicatum, and provided an improved diagnosis of Myoschiston. In addition, the coding regions for nuclear small subunit (SSU) rRNA and internal transcribed spacer 1‐5.8S‐internal transcribed spacer 2 sequences were sequenced. Phylogenetic analyses that included available SSU rDNA sequences of peritrichs from GenBank strongly supported a position of M. duplicatum within the family Zoothamniidae. In addition, phylogenetic analyses were performed with single datasets (ITS1‐5.8S‐ITS2) and combined datasets (SSU rDNA + ITS1‐5.8S‐ITS2) to explore further the phylogenetic relationship in the family Zoothamniidae between the three morphologically similar genera—Zoothamnium, Myoschiston, and Zoothamnopsis.     

An ITS-based phylogenetic framework for the genus Vorticella: finding the molecular and morphological gaps in a taxonomically difficult group

Vorticella includes more than 100 currently recognized species and represents one of the most taxonomically challenging genera of ciliates. Molecular phylogenetic analysis of Vorticella has been performed so far with only sequences coding for small subunit ribosomal RNA (SSU rRNA); only a few of its species have been investigated using other genetic markers owing to a lack of similar sequences for comparison. Consequently, phylogenetic relationships within the genus remain unclear, and molecular discrimination between morphospecies is often difficult because most regions of the SSU rRNA gene are too highly conserved to be helpful. In this paper, we move molecular systematics for this group of ciliates to the infrageneric level by sequencing additional molecular markers—fast-evolving internal transcribed spacer (ITS) regions—in a broad sample of 66 individual samples of 28 morphospecies of Vorticella collected from Asia, North America and Europe. Our phylogenies all featured two strongly supported, highly divergent, paraphyletic clades (I, II) comprising the morphologically defined genus Vorticella. Three major lineages made up clade I, with a relatively well-resolved branching order in each one. The marked divergence of clade II from clade I confirms that the former should be recognized as a separate taxonomic unit as indicated by SSU rRNA phylogenies. We made the first attempt to elucidate relationships between species in clade II using both morphological and multi-gene approaches, and our data supported a close relationship between some morphospecies of Vorticella and Opisthonecta, indicating that relationships between species in the clade are far more complex than would be expected from their morphology. Different patterns of helix III of ITS2 secondary structure were clearly specific to clades and subclades of Vorticella and, therefore, may prove useful for resolving phylogenetic relationships in other groups of ciliates.     

Molecular classification and phylogenetic relationships of selected edible Basidiomycetes species

Morphological identification of edible mushrooms can sometimes prove troublesome, because phenotypic variation in fungi can be affected by substrate and environmental factors. One of the most important problems for mushroom breeders is the lack of a systematic consensus tool to distinguish different species, which are sometimes morphologically identical. Basidiomycetes as one of the largest groups of edible mushrooms have become more important in recent times for their medicinal and nutritional properties. Partial rDNA sequences, including the Internal Transcribed Spacer I-5.8SrDNA-Internal Transcribed Spacer II, were used in this study for molecular identification and assessment of phylogenetic relationships between selected edible species of the Basidiomycetes. Phylogenetic trees showed five distinct clades; each clade belonging to a separate family group. The first clade included all the species belonging to the Pleurotaceae (Pleurotus spp.) family; similarly, the second, third, fourth, and fifth clades consist of species from the Agaricaceae (Agaricus sp.), Lyophllaceae (Hypsigygus sp.), Marasmiaceae (Lentinula edodes sp.) and Physalacriaceae (Flammulina velutipes sp.) families, respectively. Moreover, different species of each family were clearly placed in a distinct sub-cluster and a total of 13 species were taken for analysis. Species differentiation was re-confirmed by AMOVA analysis (among the populations: 99.67%; within: 0.33%), nucleotide divergence, haplotyping and P value. Polymorphism occurred throughout the ITS regions due to insertion-deletion and point mutations, and can be clearly differentiated within the families as well as genera. Moreover, this study proves that the sequence of the ITS region is a superior molecular DNA barcode for taxonomic identification of Basidiomycetes.     

Wild Mushroom Markets in Central Mexico and a Case Study at Ozumba

Wild Mushroom Markets in Central Mexico and a Case Study at Ozumba. More than 200 species of wild edible mushrooms are consumed in Mexico, and over 100 are sold in local, regional, and wholesale markets. This paper analyzes general trends in the commercialization of wild edible mushrooms at twelve markets in central Mexico, where at least 90 species are sold and more than 100 local names were registered. The paper then focuses its analysis on the traditional market at Ozumba (state of Mexico) near Izta-Popo and Zoquiapan national parks. In 2006, 411 stands selling more than 60 mushroom species were sampled throughout the year to record information on the vendors and the species they sold. The greatest diversity of species was recorded in July and August. More than 90% of the wild mushroom vendors were women, and 64% were 40 to 60 years old. The economic value of wild mushrooms in some regions of Mexico appears to contribute to the maintenance of traditional ethnobiological knowledge, generally observed to be in overall decline.     

Morphological and molecular taxonomy of novel species <Emphasis Type="Italic">Pleurotheciaceae</Emphasis> from freshwater habitats in Yunnan,China

The diversity of lignicolous freshwater fungi of the Greater Mekong Subregion are currently being studied. In this paper, 18 fresh collections of Pleurotheciaceae from submerged wood in freshwater are characterized based on morphological characters and analyses of ITS, LSU, SSU and RPB2 sequence data. Six new species of Pleurotheciella (P. aquatica, P. fusiformis, P. guttulata, P. lunata, P. saprophytica, P. submersa), one new Phaeoisaria species (Ph. aquatica) and one new Pleurothecium species (Pl. aquaticum) are introduced based on their distinct morphology and evidence from molecular phylogeny. Pleurotheciella uniseptata, Phaeoisaria clematidis and Pleurothecium pulneyense are also redescribed and phylogenetic relationships assessed herein, these species are the first records for China.     

Escaping introns in COI through cDNA barcoding of mushrooms: Pleurotus as a test case

DNA barcoding involves the use of one or more short, standardized DNA fragments for the rapid identification of species. A 648‐bp segment near the 5′ terminus of the mitochondrial cytochrome c oxidase subunit I (COI) gene has been adopted as the universal DNA barcode for members of the animal kingdom, but its utility in mushrooms is complicated by the frequent occurrence of large introns. As a consequence, ITS has been adopted as the standard DNA barcode marker for mushrooms despite several shortcomings. This study employed newly designed primers coupled with cDNA analysis to examine COI sequence diversity in six species of Pleurotus and compared these results with those for ITS. The ability of the COI gene to discriminate six species of Pleurotus, the commonly cultivated oyster mushroom, was examined by analysis of cDNA. The amplification success, sequence variation within and among species, and the ability to design effective primers was tested. We compared ITS sequences to their COI cDNA counterparts for all isolates. ITS discriminated between all six species, but some sequence results were uninterpretable, because of length variation among ITS copies. By comparison, a complete COI sequences were recovered from all but three individuals of Pleurotus giganteus where only the 5′ region was obtained. The COI sequences permitted the resolution of all species when partial data was excluded for P. giganteus. Our results suggest that COI can be a useful barcode marker for mushrooms when cDNA analysis is adopted, permitting identifications in cases where ITS cannot be recovered or where it offers higher resolution when fresh tissue is. The suitability of this approach remains to be confirmed for other mushrooms.     

Mushroom diversity in sustainable shade tea forest and the effect of fire damage

A survey of the biodiversity of wild macrofungi, including edible species yields, was carried out from 1 May to 30 September 2007 at four different forest types (in mainly Miang tea forest). The plots 100 m², comprised a tea garden with a few planted canopy tree species (37.2% canopy cover), a cultivated sustainable tea forest (80.2% canopy cover), an abandoned sustainable tea forest (89.8% canopy cover), and an abandoned sustainable tea forest that had suffered fire damage (72.9% canopy cover). All visible mushrooms were collected during weekly forays. Macro-characters of the fungi were annotated and photographed by digital camera and the fungi were identified to genera and morphospecies (e.g. Agaricus sp.1, Agaricus sp.2). The fresh weight of wild edible mushrooms produced in the plots was also recorded during this period. The biodiversity of macrofungi in abandoned sustainable tea forest was highest with 115 species in 47 genera, followed by cultivated sustainable tea forest with 85 species in 42 genera and fire damaged abandoned sustainable tea forest with 48 species in 25 genera, while only 19 species belonging to 15 genera were found in the tea garden. Twenty-one species belonging to nine genera with a total of 60.9 kg of edible fungi were collected throughout this study. The fresh weight of edible mushrooms was recorded with the highest fresh weight (25.3 kg) collected from the abandoned sustainable tea forest, followed by 18.2 kg in the cultivated sustainable tea forest, while in the fire damaged sustainable tea forest 16.4 kg were collected and only 1 kg was collected in the tea garden. Abandoned sustainable tea forest contained a greater shade tree biodiversity and higher canopy cover than other plots. This area has a generally higher humidity, a greater ground litter cover, a greater number of microhabitats, a greater number of trees that can form mycorrhizal associations and this probably accounts for the higher diversity of macrofungi and production of edible mushrooms. The fire damaged sustainable tea forest had lower mushroom diversity and edible mushroom production possibly due to the loss of litter and lower tree diversity. The tea garden supported a poor diversity of mushrooms and almost no edible mushrooms. Shade tea forest (Miang tea forest) is a good method to produce tea in a sustainable way as it maintains diversity of mushrooms and other organisms and could be developed as an alternative to shade coffee.     

Morphological and molecular characterization of Tulasnella spp. fungi isolated from the roots of Epidendrum secundum,a widespread Brazilian orchid

Tulasnella spp. are the main fungal symbionts of Brazilian Epidendrum orchids. The taxonomy of these fungi is largely based on ITS rDNA similarity, but culture dependent techniques are still essential to establish the true biological entity of the mycobiont. The aim of this study was to characterize morphologically and molecularly 16 Tulasnella spp. fungi isolated from three different populations of E. secundum and to test the coincidences between morphological and molecular characterization. Two uninucleate rhizoctonia fungi, obtained from Oncidium barbaceniae, and two phytopathogenic isolates were included as outgroups. Qualitative and quantitative morphological characteristics were analyzed using multivariate statistics and were able to distinguish Ceratobasidium, Tulasnella and Thanatephorus genera and separate the isolates of Tulasnella spp. into two groups. Analysis of RAPD (Random Amplified Polymorphic DNA) and ITS rDNA sequences validated the morphological data. Symbionts of O. barbaceniae presented identity to ITS sequences of Ceratobasidium genus, while E. secundum isolates presented identity to two species of Tulasnella. We observed homogeneity among Tulasnella spp. obtained from a single population and from neighboring populations, but there was higher variability among isolates obtained from populations of regions that were farther apart. Morphological data associated with multivariate statistics proved to be a useful tool in the multi-level taxonomy of these orchid-associated fungi and in estimating the diversity of orchid mycorrhizal fungi.     

Three New Species of Cyphellophora (Chaetothyriales) Associated with Sooty Blotch and Flyspeck

The genus Cyphellophora includes human- and plant-related species from mammal skin and nails, plant materials, and food. On the basis of analysis of ITS, LSU, TUB2 and RPB1 data and morphological characters, three new species, Cyphellophora phyllostachysdis, C. artocarpi and C. musae, associated with sooty blotch and flyspeck disease, were added to this genus. The 2D structure of ITS1 and ITS2 confirmed this taxonomic status. Pathogenicity tests on apple fruit indicated that C. artocarpi could be a sooty blotch and flyspeck pathogen of apple.     

Diversity of macrofungi and exploitation of edible mushroom resources in the National Park “Appennino Lucano,Val D'Agri,Lagonegrese” (Italy)

An investigation on the macrofungal diversity of the National Park “Appennino Lucano, Val D'Agri, Lagonegrese” (Basilicata, southern Italy) was carried out, together with an evaluation of wild edible mushrooms in agro-forest ecosystems and their possible exploitation as a new source of food and revenue in rural and under-developed areas of this region. An unpublished list of 249 mushroom taxa (229 Basidiomycota and 20 Ascomycota), mostly belonging to the genera Tricholoma, Tuber, Russula, Amanita, Boletus, Lactarius, Mycena, Agaricus, Clitocybe and Hygrophorus, growing in this important natural area is provided together with ecological characterization of the recorded specimens. Inonotus obliquus is a new record for Basilicata, while Mycena bresadolana is reported for the first time from southern Italy. The presence of Amaurodon mustialaënsis, an extremely rare species of the northern hemisphere reported for the first time on white fir, is noteworthy. Among the previous findings, 116 species of edible mushrooms are presented and discussed in view of their relative abundance and edibility.