Brown rot decay of copper-chromated-phosphorus impregnated fence poles: Characterization by molecular analyses and microscopy

At one location in central Sweden, agricultural pine (Pinus sylvestris L.) fence poles treated with a commercial copper–chromium–phosphorus preservative (CCP) formulation according to use class 4 at retention of 30 kg m⁻³ were prematurely degraded by fungi after only two years in-service. Light- and electron microscopy analyses showed decay to result from primarily brown rot attack. Culture studies produced on different agar and copper-containing media using small wood slivers removed from infected poles allowed establishment of a number of pure cultures of Phycomycetes, Ascomycetes, Fungi Imperfecti and Basidiomycete fungi. Using morphological characters, PCR and sequencing of isolated strains, Antrodia vaillantii was determined as the most abundant basidiomycete present and as the major causal agent of decay. Compatibility tests and comparison of the ITS nrDNA sequences of our putative A. vaillantii isolate with other A. vaillantii strains and with Antrodia radiculosa showed differences suggesting a hybrid strain. A combination of site characteristics (e.g. hot spots of A. vaillantii), the use of juvenile poles, copper tolerance and overall ineffectivity of CCP against A. vaillantii is suggested as reasons for premature decay.     

Antrodia onychoides (Egel.) Ryv. specie nuova per l'Italia. II

Abstract

Antrodia onychoides (Egel.) Ryv., new to Italy and very rare species in Europe. II. - The paper is concerned with the first Italian collection of A. onychoides, a resupinate polypore. From the literature, this is the fourth European finding of Antrodia onychoides and follows the first collection, occurred in 1913 near Oslo, in Norway, the second in France and the third in Finland.

Macroscopically A. onychoides could be misdetermined as a Tyromyces however, on the basis of microscopical features, there is no doubt concerning its determination as a species of Antrodia genus.     

Antrodia hyalina, a new polypore from Russia, and A. leucaena, new to Europe

A new polypore species, Antrodia hyalina, is described from Russia. It is morphologically similar to Antrodia pulvinascens, but differs in having annual, thinner and softer basidiocarps, solid skeletal hyphae, and cylindrical spores. Antrodia leucaena, originally described from China, is reported as new from Finland and Russia on Populus tremula. Antrodia wangii is regarded as a synonym of A. bondartsevae.     

东喜马拉雅地区多孔菌区系和生态习性

对东喜马拉雅地区多孔菌区系和生态习性进行了分析,发现该地区多孔菌极为丰富,共有10目19科101属372种,其中优势科为多孔菌科和锈革孔菌科,优势属为木层孔菌属和多孔菌属。种的区系地理成分可以分为8类,其中北温带类型种类最多,表明东喜马拉雅地区多孔菌具有明显的北温带区系特征。该地区的多孔菌有常见种、偶见种、稀有种和濒危种分别为156、141、54和21个;有腐生菌304种、寄生菌49种、菌根菌19种。在寄主方面,能够生长在被子植物上的有256种,裸子植物上的有137种,既能生长在被子植物,也能生长在裸子植物上的有40种,地上生长的有19种。东喜马拉雅地区的裸子植物,特别是云杉属、松属和冷杉属树木对于多孔菌的生长更重要,因为生长在这些属树木上的多孔菌分别有71、68和51种,高于该地区被子植物其他属树木上的多孔菌。     

Hidden diversity in the <Emphasis Type="Italic">Antrodia malicola</Emphasis> group (Polyporales,Basidiomycota)

Taxonomy of the Antrodia malicola group is revised based on DNA, morphological, ecological, and geographic data. This species complex is not related to Antrodia s. str. but constitutes its own lineage within the large Fomitopsis – Daedalea clade. The A. malicola group includes five species. Antrodia malicola s. str. is distributed in North America and East Asia, and a few records of this species are reported from Azores and Africa. Its European counterpart is A. kuzyana, comb. nova, with wider pores. Infraspecific variability of A. malicola and possible gene flow between it and A. kuzyana in East Asia are detected based on tef1 sequence data. Antrodia cyclopis, sp. nova, is described as a large-spored relative of A. malicola from New Guinea. Antrodia minuta is reported here from several European countries and Siberia, and its morphological characters and host preferences are discussed. Its close relative is A. tuvensis, sp. nova, from Central Siberia, an unusual member of the group with sparse, flexuous skeletal hyphae.     

Phylogenetic analysis of Antrodia and related taxa based on partial mitochondrial SSU rDNA sequences

Sequences of mitochondrial SSU rDNA were obtained from six species of Antrodia and related fungal taxa to reveal their phylogenetic relationships. Phylogenetic analysis showed that the species of Antrodia did not cluster into a single clade. Brown rot fungi were separated into two main groups through which Antrodia was dispersed. Antrodia sinuosa, A. serialis, A. heteromorpha and A. malicola clustered with Perenniporia, Fomitopsis, Piptoporus, Daedalea and Melanoporia within one group of brown rot fungi, while A. carbonica and A. vaillantii clustered with Oligoporus, Gloeophyllum and Auriporia within the other group of brown rot fungi, indicating that Antrodia is a heterogeneous genus and that brown rot fungi have evolved convergently. This revised version was published online in June 2006 with corrections to the Cover Date.     

双向电泳和质谱技术分析樟芝无性孢子萌发相关蛋白

【目的】采用双向电泳(2DE)、质谱技术和实时荧光定量PCR(RT-q PCR)技术分析樟芝无性孢子萌发相关蛋白。【方法】分别提取培养0 h和24 h的樟芝孢子总蛋白并进行双向电泳分离,再用PDQuest软件进行差异蛋白分析,并用MALDI-TOF-MS技术对差异蛋白进行鉴定;其次将鉴定成功的蛋白与孢子萌发相关蛋白的本地数据库进行比对,获得樟芝中的孢子萌发相关蛋白信息,最后用RT-qPCR技术对相关基因的转录水平进行分析。【结果】两组样品共有32个差异蛋白点,其中在24 h表达量上调的蛋白25个,下调的蛋白7个。将32个差异蛋白点挖取鉴定,成功鉴定24个。其中,与孢子萌发相关的蛋白有10个,分别为GerO、Ubc1、Cat-1、Snf1、Cas2、SfaD、Chaperonin、Fad5、Tyrosine-P和ChiA。【结论】该研究结果为进一步解析樟芝无性孢子萌发的分子机制提供了理论依据。     

Antrodia tropica sp. nov. from southern China inferred from morphological characters and molecular data

Antrodia tropica sp. nov. is described and illustrated on the basis of collections originating from Hainan, southern tropical China. Both the morphology and phylogenetic analysis of rDNA ITS sequences support this new species. Morphologically, it is characterized by resupinate basidiocarps, an annual growth habit, greyish to pinkish buff pore surface, a dimitic hyphal system with clamped generative hyphae, and cylindrical to subfusiform basidiospores. The hymenophoral trama is dominated by generative hyphae, whereas skeletal hyphae are dominant in the subiculum. Molecular phylogeny inferred from ITS sequence data suggested a close relationship between A. tropica and two other Antrodia species, including A. huangshanensis reported from China and A. ramentacea found mostly in boreal Eurasia.     

<Emphasis Type="Italic">Antrodia multiformis</Emphasis> and <Emphasis Type="Italic">A. tenerifensis</Emphasis> spp. nov. (Fomitopsidaceae,Basidiomycota): new brown rot polypores

Based on molecular evidence and morphological studies, the new polypores Antrodia multiformis and A. tenerifensis are described. Both species were discovered in arid biotope, A. multiformis in southwestern USA and A. tenerifensis in Tenerife (Canary Islands). Both of the new species show unique morphological and cultural features and are different in phylogeny. The structure of the Antrodia core clade of brown rot polypores is discussed.     

Phylogenetic analysis of <Emphasis Type="Italic">Antrodia</Emphasis> species and <Emphasis Type="Italic">Antrodia camphorata</Emphasis> inferred from internal transcribed spacer region

The species of Antrodia are one of the difficult-to-classify and obscure groups of poroid Aphyllophorales based on morphological appearance. However, it is becoming increasingly important to reliably identify the entire suite of Antrodia camphorata strains and Antrodia species due to the potential pharmaceutical value of their biologically active ingredients. In this study, the internal transcribed spacer (ITS) region of the ribosomal RNA gene (rDNA) was sequenced and phylogenetically analyzed in a number of Antrodia fungal species and strains. ITS amplicons from the Antrodia species tested ranged in size from 543 to 610 bp; the size of the ITS of A. camphorata strains ranged from 592 to 596 bp. The overall sizes of ITS2 and 5.8S ribosomal RNA gene of all A. camphorata strains tested in this study were shown to be 217 and 158 bp, respectively. A phylogenetic analysis of ITS data generated, which included sequences of 11 A. camphorata strains and nine other Antrodia species, showed three clearly distinct groups. Group 1 includes A. camphorata, Antrodia salmonea, and Antrodia carbinca strains. Within Group 2, Antrodia sinuosa and Antrodia xantha were clustered together. Group 3 contained Antrodia albida, A. heteromorpha, A. serialis, and A. malicola. The observed sequence diversity among ITS alleles provided an effective tool for differentiating strains of A. camphorata, A. salmonea, A. xantha, A. sinuosa, or A. serialis. Polymorphisms arising within the ITS1-5.8S-ITS2 region can provide practical markers for establishing a foundation for the further expansion of an ITS sequence database of medically important fungi.     

Production and regulation of lignocellulose-degrading enzymes of <Emphasis Type="Italic">Poria</Emphasis>-like wood-inhabiting basidiomycetes

The wood-decomposing fungal species Antrodia macra, A. pulvinascens, Ceriporiopsis aneirina, C. resinascens and Dichomitus albidofuscus were determined for production of laccase (LAC), Mn peroxidase (MnP), lignin peroxidase (LiP), endo-l,4-P-β-glucanase, endo-l,4-β-xylanase, cellobiohydrolase, 1,4-β-glucosidase and 1,4-β-xylosidase. The results confirmed the brown-rot mode of Antrodia spp. which did not produce the activity of LAC and MnP. The remaining species performed detectable activity of both enzymes while no strain produced LiP. Significant inhibition of LAC production by high nitrogen was found in all white-rot species while only MnP of D. albidofuscus was regulated in the same way. The endoglucanase and endoxylanase activities of white-rotting species were inhibited by glucose in the medium while those of Antrodia spp. were not influenced by glucose concentration. The regulation of enzyme activity and bio-mass production can vary even within a single fungal genus.     

牛樟芝非核糖体多肽合成酶基因(ＡcNRPS)的克隆与鉴定

胶霉毒素属于真菌天然次生代谢产物epipolythiodioxopiperazine (ETP)家族,具有免疫抑制剂、抗真菌等多种生物活性,是由非核糖体多肽合成酶(NRPSs)催化合成。从牛樟芝(Antrodia camphorata)基因组中挖掘出非核糖体多肽合成酶基因(AcNRPS,NCBI登录号为KX430967),克隆获取其全长cDNA,并对其进行生物信息学分析和表达谱分析。结果显示AcNRPS基因cDNA全长6 687 bp;与其DNA序列比对发现AcNRPS基因含有12 个内含子;其开放阅读框编码2 229 个氨基酸残基,BLAST比对发现其含有2 个A-C-T结构域,底物需2个氨基酸;系统发育树结果显示AcNRPS与其他合成产物为胶霉毒素的NRPS基因聚为一类,其可合成胶霉毒素类化合物;表达谱分析显示,以葡萄糖和土豆蛋白胨作为碳、氮源的培养基能够有效促进牛樟芝NRPS基因的表达。     

Characterization of a strong CCA-treated wood degrader,unknown <Emphasis Type="Italic">Crustoderma</Emphasis> species

In this study, basidiomycete isolates that possessed a strong ability to degrade chromated copper arsenate (CCA)-treated wood were characterized. These fungal isolates, which were collected from CCA-treated pine log wastes, showed no recognizable morphological properties on culture media. Nucleotide sequence analysis of the large subunit rDNA of the isolates revealed that they were one species. Based on the high sequence similarity (>95%) and close phylogenetic relationship with several known species of Crustoderma, the fungal isolates characterized in this study were classified as a Crustoderma sp. In a wood degradation test, Crustoderma isolate KUC8611 produced a remarkably higher weight loss in CCA-treated Pinus radiata (68.7%), Pseudotsuga menziesii (39.7%), and Tsuga heterophylla (38.5%) wood than other evaluated basidiomycete species, including Crustoderma flavescens and Crustoderma corneum. In addition, extracellular enzymes for cellulose and protein degradation were detected when the isolates were cultured in chromogenic media, which supports the finding that isolate KUC8611 is a wood degrader. Furthermore, an in vitro test for metal tolerance revealed that isolate KUC8611 showed strong arsenic tolerance, but that it could not tolerate copper. Finally, isolate KUC8611 produced lower amounts of oxalic acid than copper-tolerant fungi such as Fomitopsis palustris and Antrodia vaillantii. To the best of our knowledge, this is the first study to report the degradation of CCA-treated wood by a Crustoderma species.     

Wood-inhabiting,polyporoid fungi in aspen-dominated forests managed for biomass in the U.S. Lake States

To better understand the potential long-term effects of biomass harvesting on biodiversity, the polyporoid fungi community was characterized from 120 plots in four aspen-dominated forests in Minnesota. Four deadwood variables (substratum species, substratum type, decay class and diameter class) were recorded for each polyporoid species occurrence. A total of 2 358 polyporoid occurrences, representing 86 species, were recorded on 16 tree species. Eight species (Trichaptum biforme, Bjerkandera adusta, Trametes hirsuta, Phellinus tremulae, Fomes fomentarius, Irpex lacteus, Fomitopsis ochracea and Antrodia serialis) made up 67 % of occurrences. Four polyporoid species (Funalia trogii, Pycnoporellus fulgens, Rigidoporus crocatus and Skeletocutis chrysella) are potentially rare and/or threatened in the Lake States. Non-metric multidimensional scaling and rarefaction curves demonstrated that small diameter substrata (especially those <5 cm) most strongly influenced polyporoid species occurrences. Aspen-dominated systems show great potential for biomass production, but these forests also support a species-rich community of polyporoid fungi, including potentially rare species.     

Is fungal species richness and composition related to the occurrence of the old‐growth associated wood‐decaying Amylocystis lapponica?

Amylocystis lapponica (Romell) Singer is a widely distributed wood‐decaying polypore fungus found throughout the Northern Hemisphere. Despite its huge distribution range it occurs rather patchily and seems narrowly associated with old‐growth forest stands. Notably, it has been used as an ‘indicator species’, believed to reflect the long‐term presence of dead wood, naturalness of forest stands, and indirectly, species richness and possibly composition. In this study we focused on the last issue – whether or not there is a link between the occurrence of A. lapponica and the species richness and composition of other wood‐decaying fungi. Selecting log characteristics and microclimate as similar as possible, we compared 12 logs with and 12 logs without visible fruit bodies of A. lapponica to examine: 1) if visible fruit bodies corresponded with molecular identification of the mycelia, 2) if fungal species richness and composition of the substrate were related to A. lapponica occurrence, and 3) if A. lapponica was restricted to certain parts of the log. Fungal species were recorded by inspecting visible fruit bodies and by culture isolation and ITS sequencing from wood disc samples. Laboratory and field identification of A. lapponica had 71% correspondence, and mycelia were identified in two logs without visible fruit bodies. Twice as many fungal species were detected using ITS sequencing compared to fruit body identification. Total species richness was similar between the two log categories, but number of species per log was slightly higher in A. lapponica logs. Antrodia serialis (Fr.) Donk, and possibly also Fomitopsis pinicola (Sw.:Fr.) P. Karst. and Phellinus nigrolimitatus (Romell) Bourdot & Galzin, occurred more frequently in A. lapponica logs. Mycelia of A. lapponica were restricted to less decayed parts of the wood in the centre of the middle part of the logs.     

中国北京地区的多孔菌

本文报道产于中国北方北京地区的非褶菌目孔状真菌72种,并列出了每种的寄主。其中大部分种类为该地区首次报道。王氏薄孔菌为一新种,它的主要特征为子实体平伏反转,孔口表面奶油色至浅黄色,担孢子圆柱型并略弯曲,生长在李属树木上。该新种与垫状薄孔菌相似,但后者孢子较宽且是完全平伏的种类。     

Effect of anoxic conditions on wood-decay fungi treated with argon or nitrogen

The effects of low-oxygen conditions, achieved with either argon or nitrogen gas, on the viability of wood-decay fungi Coniophora puteana and Antrodia vaillantii, grown on artificial growth medium, were tested. Initial tests for viability were run after 1, 2, 3 and 4 weeks of exposure to low oxygen conditions, at oxygen levels in the vessels maintained below 10 ppm. Treatment was later extended to 10 and 16 weeks. Tests included measurements of CO₂ production and determination of the ability of fungal tissue to regenerate. The effect of anoxic conditions on the mycelia of treated fungal species was expressed as an increased time needed for regeneration or as a complete absence of growth of inocula taken from the exposed cultures. The cultures that were retarded by the low-oxygen environment consequently produced less CO₂ per hour. For C. puteana cultures, the effects of anoxic treatment became apparent in the second week of the treatment. The number of affected cultures rose steadily with the prolongation of anoxic treatment. By the 16th week of the experiment, 80% of the inocula of C. puteana did not regenerate. A. vaillantii inocula regeneration was not affected until after the fourth week of treatment. The influence of anoxic treatment on the cultures of this species was more pronounced in the test on the 10th and especially on the 16th week, when 67% of inocula did not regenerate. Argon or nitrogen gas caused the same degree of loss of viability in both the fungal species tested. In general, A. vaillantii mycelial cultures proved to be less sensitive to anoxic conditions, caused by either argon and nitrogen gas.