Glutathione transferase-like proteins encoded in genomes of yeasts and fungi: insights into evolution of a multifunctional protein superfamily

Most fungal glutathione transferases (GSTs) do not fit easily into any of the previously characterised classes by immunological, sequence or catalytic criteria. In contrast to the paucity of studies on GSTs cloned or isolated from fungal sources, a screen of databases revealed 67 GST-like sequences from 21 fungal species. Comparison by multiple sequence alignment generated a dendrogram revealing five clusters of GST-like proteins designated clusters 1, 2, EFIBgamma, Ure2p and MAK16, the last three of which have previously been related to the GST superfamily. Surprisingly, a relatively small number of fungal GSTs belong to mainstream classes and the previously-described fungal Gamma class is not widespread in the 21 species studied. Representative crystal structures are available for the EFIBgamma and Ure2p classes and the domain structures of representative sequences are compared with these. In addition, there are some "orphan" sequences that do not fit into any previously-described class, but show similarity to genes implicated in fungal biosynthetic gene clusters. We suggest that GST-like sequences are widespread in fungi, participating in a wide range of functions. They probably evolved by a process similar to domain "shuffling".     

The importance of sourcing enzymes from non-conventional fungi for metabolic engineering and biomass breakdown

A wealth of fungal enzymes has been identified from nature, which continue to drive strain engineering and bioprocessing for a range of industries. However, while a number of clades have been investigated, the vast majority of the fungal kingdom remains unexplored for industrial applications. Here, we discuss selected classes of fungal enzymes that are currently in biotechnological use, and explore more basal, non-conventional fungi and their underexploited biomass-degrading mechanisms as promising agents in the transition towards a bio-based society. Of special interest are anaerobic fungi like the Neocallimastigomycota, which were recently found to harbor the largest diversity of biomass-degrading enzymes among the fungal kingdom. Enzymes sourced from these basal fungi have been used to metabolically engineer substrate utilization in yeast, and may offer new paths to lignin breakdown and tunneled biocatalysis. We also contrast classic enzymology approaches with emerging ‘omics’-based tools to decipher function within novel fungal isolates and identify new promising enzymes. Recent developments in genome editing are expected to accelerate discovery and metabolic engineering within these systems, yet are still limited by a lack of high-resolution genomes, gene regulatory regions, and even appropriate culture conditions. Finally, we present new opportunities to harness the biomass-degrading potential of undercharacterized fungi via heterologous expression and engineered microbial consortia.     

Genome-Wide Identification of Mitogen-Activated Protein Kinase Gene Family across Fungal Lineage Shows Presence of Novel and Diverse Activation Loop Motifs

The mitogen-activated protein kinase (MAPK) is characterized by the presence of the T-E-Y, T-D-Y, and T-G-Y motifs in its activation loop region and plays a significant role in regulating diverse cellular responses in eukaryotic organisms. Availability of large-scale genome data in the fungal kingdom encouraged us to identify and analyse the fungal MAPK gene family consisting of 173 fungal species. The analysis of the MAPK gene family resulted in the discovery of several novel activation loop motifs (T-T-Y, T-I-Y, T-N-Y, T-H-Y, T-S-Y, K-G-Y, T-Q-Y, S-E-Y and S-D-Y) in fungal MAPKs. The phylogenetic analysis suggests that fungal MAPKs are non-polymorphic, had evolved from their common ancestors around 1500 million years ago, and are distantly related to plant MAPKs. We are the first to report the presence of nine novel activation loop motifs in fungal MAPKs. The specificity of the activation loop motif plays a significant role in controlling different growth and stress related pathways in fungi. Hence, the presences of these nine novel activation loop motifs in fungi are of special interest.     

Microbial Pathogens in the Fungal Kingdom

The fungal kingdom is vast, spanning ~1.5 to as many as 5 million species diverse as unicellular yeasts, filamentous fungi, mushrooms, lichens, and both plant and animal pathogens. The fungi are closely aligned with animals in one of the six to eight supergroups of eukaryotes, the opisthokonts. The animal and fungal kingdoms last shared a common ancestor ~1 billion years ago, more recently than other groups of eukaryotes. As a consequence of their close evolutionary history and shared cellular machinery with metazoans, fungi are exceptional models for mammalian biology, but prove more difficult to treat in infected animals. The last common ancestor to the fungal/metazoan lineages is thought to have been unicellular, aquatic, and motile with a posterior flagellum, and certain extant species closely resemble this hypothesized ancestor. Species within the fungal kingdom were traditionally assigned to four phyla, including the basal fungi (Chytridiomycota, Zygomycota) and the more recently derived monophyletic lineage, the dikarya (Ascomycota, Basidiomycota). The fungal tree of life project has revealed that the basal lineages are polyphyletic, and thus there are as many as eight to ten fungal phyla. Fungi that infect vertebrates are found in all of the major lineages, and virulence arose multiple times independently. A sobering recent development involves the species Batrachochytrium dendrobatidis from the basal fungal phylum, the Chytridiomycota, which has emerged to cause global amphibian declines and extinctions. Genomics is revolutionizing our view of the fungal kingdom, and genome sequences for zygomycete pathogens (Rhizopus, Mucor), skin-associated fungi (dermatophytes, Malassezia), and the Candida pathogenic species clade promise to provide insights into the origins of virulence. Here we survey the diversity of fungal pathogens and illustrate key principles revealed by genomics involving sexual reproduction and sex determination, loss of conserved pathways in derived fungal lineages that are retained in basal fungi, and shared and divergent virulence strategies of successful human pathogens, including dimorphic and trimorphic transitions in form. The overarching conclusion is that fungal pathogens of animals have arisen repeatedly and independently throughout the fungal tree of life, and while they share general properties, there are also unique features to the virulence strategies of each successful microbial pathogen.     

Glycogen phosphorylase sequences from the amitochondriate protists, Trichomonas vaginalis, Mastigamoeba balamuthi, Entamoeba histolytica and Giardia intestinalis

Glycogen phosphorylase genes or messages from four amitochondriate eukaryotes, Trichomonas vaginalis, Mastigamoeba balamuthi, Entamoeba histolytica (two genes) and Giardia intestinalis, have been isolated and sequenced. The sequences of the amitochondriate protist enzymes appear to share a most recent common ancestor. The clade containing these sequences is closest to that of another protist, the slime mold (Dictyostelium discoideum), and is more closely related to fungal and plant phosphorylases than to mammalian and eubacterial homologs. Structure-based amino acid alignment shows conservation of the residues and domains involved in catalysis and allosteric regulation by glucose 6-phosphate but high divergence at domains involved in phosphorylation-dependent regulation and AMP binding in fungi and animals. Protist phosphorylases, as their prokaryotic and plant counterparts, are probably not regulated by phosphorylation.     

A matter of structure: structural comparison of fungal carbonic anhydrases

Carbonic anhydrases (CAs) are metalloenzymes that catalyze the interconversion of carbon dioxide (CO₂) and hydrogen carbonate. CAs are distributed over all the three domains of life and are divided into five distinct evolutionarily unrelated gene families (α, β, γ, δ, ζ). In the large fungal kingdom, the majority of fungi encode multiple copies of β-CAs, with some also possessing genes for α-class CAs. Hemiascomycetous and basidiomycetous yeasts encode one or two β-CAs, while most of the filamentous ascomycetes have multiple copies of genes encoding α- and β-CAs. The functions of fungal β-CAs have been investigated intensively, while the role of fungal α-CAs is mostly unknown. The β-CAs are involved in sexual development, CO₂-sensing, pathogenicity, and survival in ambient air. Only recently, researchers have begun to use functional and structural data of CAs from pathogenic and non-pathogenic organisms to develop powerful and effective drugs and inhibitors or to identify enzymes that can be utilized in industrial applications. Despite the large number of fungal CAs known, only five have been characterized structurally: the α-CA AoCA of Aspergillus oryzae, the full length β-CA Can2 from the pathogenic basidiomycete Cryptococcus neoformans, the N-terminally truncated Saccharomyces cerevisiae β-CA Nce103, and two β-CAs of Sordaria macrospora. This review focuses on the functional and structural properties of fungal CAs.     

Hybrid histidine kinases in pathogenic fungi

Histidine kinases (HK) sense and transduce via phosphorylation events many intra‐ and extracellular signals in bacteria, archaea, slime moulds and plants. HK are also widespread in the fungal kingdom, but their precise roles in the regulation of physiological processes remain largely obscure. Expanding genomic resources have recently given the opportunity to identify uncharacterised HK family members in yeasts and moulds and now allow proposing a complex classification of Basidiomycota, Ascomycota and lower fungi HK. A growing number of genetic approaches have progressively provided new insight into the role of several groups of HK in prominent fungal pathogens. In particular, a series of studies have revealed that members of group III HK, which occur in the highest number of fungal species and contain a unique N‐terminus region consisting of multiple HAMP domain repeats, regulate morphogenesis and virulence in various human, plant and insect pathogenic fungi. This research field is further supported by recent shape‐function studies providing clear correlation between structural properties and signalling states in group III HK. Since HK are absent in mammals, these represent interesting fungal target for the discovery of new antifungal drugs.     

Characterization of ATDRG1, a member of a new class of GTP-binding proteins in plants

We report the initial characterization of an Arabidopsis thaliana cDNA (atdrg1), a member of a new class of GTP-binding proteins (G-proteins) in plants. The predicted ATDRG1 protein contains all five structural motifs characteristic of the G-protein superfamily. Apart from these motifs, the amino acid sequence differs substantially from all known G-proteins except for a recently discovered new family named developmentally regulated G-proteins (DRGs). Sequences closely related to atdrg1 are found in species as distant as human (80% amino acid conservation), Drosophila (74%), yeast (77%) and Caenorhabditis elegans (77%). The remarkable evolutionary conservation of these proteins suggests an important, but as yet unclear role. Phylogenetic analysis of the available homologous sequences strongly suggests a diphyletic origin of the eukaryotic DRG proteins. Northern analysis shows high levels of atdrg1 mRNA in all Arabidopsis tissues studied, and homologues of atdrg1 are present throughout the plant kingdom. In situ hybridization reveals that atdrg1 is highly expressed in actively growing tissues and reproductive organs. Southern analysis indicates the presence of either one or two copies of atdrg1 in the Arabidopsis genome. Immunolocalization studies show that the protein is present in cytoplasmic vesicles found mainly in actively growing tissues suggesting a putative role for ATDRG1 in either the regulation of vesicle transport or the regulation of enzymes involved in storage protein processing.     

A new promising phylogenetic marker to study the diversity of fungal communities: The Glycoside Hydrolase 63 gene

In molecular ecology, the development of efficient molecular markers for fungi remains an important research domain. Nuclear ribosomal internal transcribed spacer (ITS) region was proposed as universal DNA barcode marker for fungi, but this marker was criticized for Indel‐induced alignment problems and its potential lack of phylogenetic resolution. Our main aim was to develop a new phylogenetic gene and a putative functional marker, from single‐copy gene, to describe fungal diversity. Thus, we developed a series of primers to amplify a polymorphic region of the Glycoside Hydrolase GH63 gene, encoding exo‐acting α‐glucosidases, in basidiomycetes. These primers were validated on 125 different fungal genomic DNAs, and GH63 amplification yield was compared with that of already published functional markers targeting genes coding for laccases, N‐acetylhexosaminidases, cellobiohydrolases and class II peroxidases. Specific amplicons were recovered for 95% of the fungal species tested, and GH63 amplification success was strikingly higher than rates obtained with other functional genes. We downloaded the GH63 sequences from 483 fungal genomes publicly available at the JGI mycocosm database. GH63 was present in 461 fungal genomes belonging to all phyla, except Microsporidia and Neocallimastigomycota divisions. Moreover, the phylogenetic trees built with both GH63 and Rpb1 protein sequences revealed that GH63 is also a promising phylogenetic marker. Finally, a very high proportion of GH63 proteins was predicted to be secreted. This molecular tool could be a new phylogenetic marker of fungal species as well as potential indicator of functional diversity of basidiomycetes fungal communities in term of secretory capacities.     

Virulence genes and the evolution of host specificity in plant-pathogenic fungi

In the fungal kingdom, the ability to cause disease in plants appears to have arisen multiple times during evolution. In many cases, the ability to infect particular plant species depends on specific genes that distinguish virulent fungi from their sometimes closely related nonvirulent relatives. These genes encode host-determining "virulence factors," including small, secreted proteins and enzymes involved in the synthesis of toxins. These virulence factors typically are involved in evolutionary arms races between plants and pathogens. We briefly summarize current knowledge of these virulence factors from several fungal species in terms of function, phylogenetic distribution, sequence variation, and genomic location. Second, we address some issues that are relevant to the evolution of virulence in fungi toward plants; in particular, horizontal gene transfer and the genomic organization of virulence genes.     

古田山国家级自然保护区木腐真菌物种多样性及分布

木腐真菌是微生物的一个重要类群, 主要以倒木为生长基质, 通过产生各种水解酶将倒木的纤维素、木质素和半纤维素分解为小分子物质, 对促进森林生态系统中的营养物质循环发挥着重要的生态功能。于2016年8月在浙江古田山国家级自然保护区开展的木腐真菌野外调查, 利用形态学和DNA序列分析对采集的标本进行了物种鉴定, 并分析了木腐真菌的物种组成和地理成分。在采集的158份标本中鉴定木腐真菌45属92种, 其中白腐真菌78种, 褐腐真菌14种。古田山的木腐真菌物种区系组成中, 热带-亚热带成分比例最高。在158份木腐真菌标本中, 97份标本采自直径大于10 cm的倒木或树桩上, 分属于76个种, 是木腐真菌生长的主要基质大小类型; 48份标本采自直径为2-10 cm的枝干上, 分属38个种; 13份标本采自直径小于2 cm的枝干上, 分属12种。不同腐烂等级倒木上生长的真菌数量和种类差异明显, 其中一级腐烂倒木上采集到9份标本(7种), 二级腐烂倒木上采集到86份标本(45种), 三级腐烂倒木上49份标本(29种), 四级腐烂倒木上14份标本(14种)。结果表明, 林分中倒木直径大小和腐烂程度是影响木腐真菌生长与分布的重要因子。     

Formins: intermediates in signal-transduction cascades that affect cytoskeletal reorganization

The control of cell growth and polarity depends on a dynamic actin cytoskeleton that has the ability to reorganize in response to developmental and environmental stimuli. In animals and fungi, formins are just one of the four major classes of poly-L-proline-containing (PLP) proteins that form part of the signal-transduction cascade that leads to rearrangement of the actin cytoskeleton. Analysis of the Arabidopsis genome sequence indicates that, unlike animals and fungi, formins are the only class of conserved profilin-binding PLP proteins in plants. Moreover, plant formins show significant structural differences compared with their animal and fungal counterparts, raising the possibility that plant formins are subject to novel mechanisms of control or perform unique roles in plants.     

Comparative Genome Analysis Reveals an Absence of Leucine-Rich Repeat Pattern-Recognition Receptor Proteins in the Kingdom Fungi

Background

In plants and animals innate immunity is the first line of defence against attack by microbial pathogens. Specific molecular features of bacteria and fungi are recognised by pattern recognition receptors that have extracellular domains containing leucine rich repeats. Recognition of microbes by these receptors induces defence responses that protect hosts against potential microbial attack.

Methodology/Principal Findings

A survey of genome sequences from 101 species, representing a broad cross-section of the eukaryotic phylogenetic tree, reveals an absence of leucine rich repeat-domain containing receptors in the fungal kingdom. Uniquely, however, fungi possess adenylate cyclases that contain distinct leucine rich repeat-domains, which have been demonstrated to act as an alternative means of perceiving the presence of bacteria by at least one fungal species. Interestingly, the morphologically similar osmotrophic oomycetes, which are taxonomically distant members of the stramenopiles, possess pattern recognition receptors with similar domain structures to those found in plants.

Conclusions

The absence of pattern recognition receptors suggests that fungi may possess novel classes of pattern-recognition receptor, such as the modified adenylate cyclase, or instead rely on secretion of anti-microbial secondary metabolites for protection from microbial attack. The absence of pattern recognition receptors in fungi, coupled with their abundance in oomycetes, suggests this may be a unique characteristic of the fungal kingdom rather than a consequence of the osmotrophic growth form.     

Yeast chitin synthases 1 and 2 consist of a non-homologous and dispensable N-terminal region and of a homologous moiety essential for function

Predicted protein sequences of fungal chitin synthases can be divided into a non-homologous N-terminal region and a C-terminal region that shows significant homology among the various synthases. We have explored the function of these domains by constructing a series of nested deletions, extending from either end, in theCHS1 andCHS2 genes ofSaccharomyces cerevisiae. In both cases, most or all of the sequences encoding the non-homologous N-terminal region (one-third of the protein for Chs1p and about one-fourth for Chs2p) could be excised, with little effect on the enzymatic activity in vitro of the corresponding synthase or on its function in vivo. However, further small deletions (20–25 amino acids) into the homologous region were deleterious to enzymatic activity and function, and often led to changes in the zymogenic character of the enzymes. Similarly, relatively small (about 75 amino acids) deletions from the C-terminus resulted in loss of enzymatic activity and function of both synthases. Thus, it appears that all the information necessary for membrane localization, enzymatic activity and function resides in the homologous regions of Chs1p and Chs2p, a situation that may also apply to other chitin synthases.These authors contributed equally to this paper     

Spitzenkorper localization and intracellular traffic of green fluorescent protein-labeled CHS-3 and CHS-6 chitin synthases in living hyphae of Neurospora crassa

The subcellular location and traffic of two selected chitin synthases (CHS) from Neurospora crassa, CHS-3 and CHS-6, labeled with green fluorescent protein (GFP), were studied by high-resolution confocal laser scanning microscopy. While we found some differences in the overall distribution patterns and appearances of CHS-3-GFP and CHS-6-GFP, most features were similar and were observed consistently. At the hyphal apex, fluorescence congregated into a conspicuous single body corresponding to the location of the Spitzenkörper (Spk). In distal regions (beyond 40 μm from the apex), CHS-GFP revealed a network of large endomembranous compartments that was predominantly comprised of irregular tubular shapes, while some compartments were distinctly spherical. In the distal subapex (20 to 40 μm from the apex), fluorescence was observed in globular bodies that appeared to disintegrate into vesicles as they advanced forward until reaching the proximal subapex (5 to 20 μm from the apex). CHS-GFP was also conspicuously found delineating developing septa. Analysis of fluorescence recovery after photobleaching suggested that the fluorescence of the Spk originated from the advancing population of microvesicles (chitosomes) in the subapex. The inability of brefeldin A to interfere with the traffic of CHS-containing microvesicles and the lack of colocalization of CHS-GFP with the endoplasmic reticulum (ER)-Golgi body fluorescent dyes lend support to the idea that CHS proteins are delivered to the cell surface via an alternative route distinct from the classical ER-Golgi body secretory pathway.Fungal hyphae elongate and branch by a complex process based on polarized secretion. Many studies have investigated the cellular and molecular components involved in shaping fungal cells, but no detailed understanding of the mechanisms that govern and regulate polarized fungal growth has been achieved (4, 25). In the yeast Saccharomyces cerevisiae, many of the main components of the secretory pathway, including some of the enzymes involved in cell wall formation, have been extensively characterized (32). Filamentous fungi encode homologues of some key components known from the yeast secretory pathway, but despite their apparent orthology, relatively little is known about how this pathway is organized to accomplish the highly polarized growth typical of hyphae. There are some differences in cell wall synthesis between filamentous fungi and S. cerevisiae. In hyphae of septate fungi, vesicles and other components accumulate at the apex, as part of the Spitzenkörper (Spk) (14, 22-24, 28). The composition and mode of action of this pleomorphic and dynamic structure have intrigued fungal biologists for many decades.Fungal cells have at least two types of well-defined secretory vesicles (5). It has been suggested that macrovesicles, or conventional secretory vesicles, carry the components of the amorphous phase of the cell wall, in addition to the load of extracellular enzymes (5, 27). There is a large body of evidence characterizing the chitin synthase (CHS)-carrying microvesicles as chitosomes (3, 8, 13, 30). CHS are β-glycosyltransferases that catalyze the polymerization of N-acetylglucosamine from UDP N-acetylglucosamine into chitin (47), a major structural polymer of the fungal cell wall (2). Chitin synthesis occurs in highly localized fashion both at the hyphal apices (7) and at nascent septa (29). Chitosomes are the smallest vesicles with the ability to form chitin microfibrils in vitro and have been suggested to carry and transport CHS to the cell surface at the apex of hyphae for cell wall synthesis (13, 37, 48, 55, 56). In recent years, studies on fungal CHS have concentrated mainly on gene identification. Given this wealth of information, we chose CHS as candidate markers to investigate vesicle traffic in fungal hyphae.Fungi have multiple chs genes grouped into two divisions, with seven classes, primarily on the basis of similarities in the primary sequence of the predicted proteins (12, 16, 37, 50). Division I includes classes I, II, and III, which share a catalytic domain surrounded by a hydrophilic N-terminal region and a hydrophobic C-terminal region (12). Division II includes classes IV, V, and VII, all with a catalytic domain preceded by a cytochrome b₅-like domain. In addition, classes V and VII contain an N-terminal myosin motor-like domain, suggesting a direct interaction with the actin cytoskeleton (15, 20, 58). Class VI has not been assigned to either division and includes recently identified CHS of unknown function (16). Earlier studies suggest that the various CHS have specific roles in chitin cell wall synthesis that are time or space dependent (60). In contrast to most filamentous fungi, S. cerevisiae (46) and Candida albicans (40) have only three or four CHS isozymes, respectively. S. cerevisiae Chs1p, C. albicans Chs2p, and C. albicans Chs8p belong to class I; S. cerevisiae Chs2p and C. albicans Chs1p belong to class II; and S. cerevisiae Chs3p and C. albicans Chs3p belong to class IV (46). While potential roles in hyphal growth have been suggested for some of the seven CHS classes described in filamentous fungi (9, 64, 65), we lack specific information on the cellular localization and trafficking to their sites of action in regions of active cell wall growth for most of these proteins.The goal of this study was to elucidate the traffic of CHS-containing vesicles en route from their site of genesis to their site of exocytosis in living hyphae of Neurospora crassa. The availability of an almost-complete genome sequence for this fungus allowed the identification of seven open reading frames with high homology to previously described chs genes (10). We chose to trace the intracellular location and secretory paths of CHS-3 and CHS-6. Neurospora CHS-3 belongs to the previously reported class I CHS with known homologues in all fungi tested, including S. cerevisiae Chs1p. In contrast, CHS-6 is a newly identified CHS assigned to class VI, homologous to Aspergillus fumigatus ChsD (39) and Coccidioides posadasii CHS-6 (34) but with no apparent homologues in S. cerevisiae or C. albicans. To trace both proteins, we fused green fluorescent protein (GFP) to the carboxyl terminus of the CHS coding regions and analyzed the fate of the resulting CHS-3-GFP and CHS-6-GFP fusion proteins by high-resolution confocal laser scanning microscopy (CLSM) in living hyphae of N. crassa.     

Die Zellwand der Pilze als Korsett

Form follows function: The fungal cell wall as a support structure Within the domain of Eukarya, the fungi form a seperate kingdom. The typical formation of branched mycelia from single hyphae is based on cell wall production at the growing hyphal tip. There, excretory vesicle fuse with the membrane releasing cell wall synthesis enzymes like chitin synthase forming the polymer of N‐acetyl glucosamin, the backbone of fungal cell walls. In addition, glucan synthases form the structural component β‐1.3‐glucan. Via β‐1,6‐glucan, cell wall proteins can be linked to the maturing cell wall, and α‐1,3‐glucan can form a matrix within the cell wall, but also a slimy matrix secreted into the medium. A layer of hydrophobins allows for growth into the air, but also facilitates formation of macroscopic structures like mushrooms.     

5种优势腐生真菌降解华山松针叶的酶活测定

【目的】分析腐生真菌降解华山松落针过程中各种酶活性变化与酶间关系,探索真菌对华山松落针的降解能力。【方法】通过形态学观察和转录间隔区ITS序列分析,鉴定分离自华山松凋落物的5种优势菌株。以此为供试菌株,华山松落针为底物,通过发酵纯培养方法,测定底物总有机物质(Total organic matter,TOM)质量损失率及在发酵过程中产生的内切葡聚糖酶(Endoglucanase,EG)、木聚糖酶(Xylanase,Xyl)、木质素过氧化物酶(Lignin peroxidase,Li P)、锰过氧化物酶(Manganese peroxidase,Mn P)和酸性磷酸酶(Acid phosphatase,AP)活性。【结果】5种菌株分别为Mucor sp.、Pestalotiopsis sp.、Allantophomopsis sp.、Phoma sp.和Hypocrea sp.。5种菌引起的针叶TOM质量损失率在6.63%-15.77%之间,Pestalotiopsis sp.具有最高的AP酶活性,且EG酶、Xyl酶和Li P酶3种酶活性较高。Allantophomopsis sp.的Li P酶活性最高,并具有很高的Mn P酶活。Hypocrea sp.分泌的EG酶、Xyl酶活性低,但能产生Li P酶且有较高的AP酶活。相关性分析表明菌株分泌的AP酶活性与TOM质量损失率负相关,EG酶、Xyl酶及AP酶3种酶之间存在协同作用,特别是EG与AP之间。【结论】5种腐生真菌对华山松针叶均有降解作用,降解能力:Pestalotiopsis sp.>Allantophomopsis sp.>Hypocrea sp.>Phoma sp.>Mucor sp.。酶活大小及酶协同作用共同影响针叶降解,Pestalotiopsis sp.、Allantophomopsis sp.和Hypocrea sp.能产生木质纤维素降解酶并能引起较高的质量损失率,因此这3株菌为木质纤维素降解真菌。     

Domain II Hairpin Structure in ITS1 Sequences as an Aid in Differentiating Recently Evolved Animal and Plant Pathogenic Fungi

The hypothesis that ITS structural features can be used to define fungal groups, where sequence analysis is unsatisfactory, was examined in plant and animal pathogenic fungi. Structural models of ITS1 regions were predicted for presumed closely related species in Colletotrichum and Trichophyton anamorphs of Arthroderma species. Structural alignment of models and comparison with ITS sequence analysis identified a variable region in a conserved hairpin formed from a common inverted repeat. Thirteen different hairpin structure models were obtained for Colletotrichum species and five different models were obtained for Trichophyton species. The different structure types could be matched to individual species and species complexes as defined by ITS sequence analysis.     

Module evolution and substrate specificity of fungal nonribosomal peptide synthetases involved in siderophore biosynthesis

Background  

Most filamentous ascomycete fungi produce high affinity iron chelators called siderophores, biosynthesized nonribosomally by multimodular adenylating enzymes called nonribosomal peptide synthetases (NRPSs). While genes encoding the majority of NRPSs are intermittently distributed across the fungal kingdom, those encoding ferrichrome synthetase NRPSs, responsible for biosynthesis of ferrichrome siderophores, are conserved, which offers an opportunity to trace their evolution and the genesis of their multimodular domain architecture. Furthermore, since the chemistry of many ferrichromes is known, the biochemical and structural 'rules' guiding NRPS substrate choice can be addressed using protein structural modeling and evolutionary approaches.