Exploiting the Biosynthetic Potency of Taxol from Fungal Endophytes of Conifers Plants; Genome Mining and Metabolic Manipulation.

Endophytic fungi have been considered as a repertoire for bioactive secondary metabolites with potential application in medicine, agriculture and food industry. The biosynthetic pathways by fungal endophytes raise the argument of acquisition of these machineries of such complex metabolites from the plant host. Diterpenoids "Taxol" is the most effective anticancer drug with highest annual sale, since its discovery in 1970 from the Pacific yew tree, Taxus brevifolia. However, the lower yield of Taxol from this natural source (bark of T. brevifolia), availability and vulnerability of this plant to unpredicted fluctuation with the ecological and environmental conditions are the challenges. Endophytic fungi from Taxus spp. opened a new avenue for industrial Taxol production due to their fast growth, cost effectiveness, independence on climatic changes, feasibility of genetic manipulation. However, the anticipation of endophytic fungi for industrial Taxol production has been challenged by the loss of its productivity, due to the metabolic reprograming of cells, downregulating the expression of its encoding genes with subculturing and storage. Thus, the objectives of this review were to (1) Nominate the endophytic fungal isolates with the Taxol producing potency from Taxaceae and Podocarpaceae; (2) Emphasize the different approaches such as molecular manipulation, cultural optimization, co-cultivation for enhancing the Taxol productivities; (3) Accentuate the genome mining of the rate-limiting enzymes for rapid screening the Taxol biosynthetic machinery; (4) Triggering the silenced rate-limiting genes and transcriptional factors to activates the biosynthetic gene cluster of Taxol.

Cytochalasans from endophytic fungus Diaporthe sp. SC-J0138.

Five new cytochalasans, diaporthichalasins D-H (1-5), along with five known cytochalasans (6-10) were isolated from solid cultures of the endophytic fungus Diaporthe sp. SC-J0138 isolated from the leaves of Cyclosorus parasiticus. Their structures were elucidated by analysis of spectroscopic data and theoretical calculations of electronic circular dichroism spectra. Compounds 1 and 5 showed noticeable cytotoxicity against human carcinoma A549, HeLa, and HepG2 cells. The structure-activity relationships in cytotoxicity were discussed for this group of compounds.

An endophytic Kocuria palustris strain harboring multiple arsenate reductase genes.

Aiming at revealing the arsenic (As) resistance of the endophytic Kocuria strains isolated from roots and stems of Sphaeralcea angustifolia grown at mine tailing, four strains belonging to different clades of Kocuria based upon the phylogeny of 16S rRNA genes were screened for minimum inhibitory concentration (MIC). Only the strain NE1RL3 was defined as an As-resistant bacterium with MICs of 14.4/0.0125 mM and 300/20.0 mM for As3+ and As5+, respectively, in LB/mineral media. This strain was identified as K. palustris based upon analyses of cellular chemical compositions (cellular fatty acids, isoprenoides, quinones, and sugars), patterns of carbon source, average nucleotide identity of genome and digital DNA-DNA relatedness. Six genes coding to enzymes or proteins for arsenate reduction and arsenite-bumping were detected in the genome, demonstrating that this strain is resistant to As possibly by reducing As5+ to As3+, and then bumping As3+ out of the cell. However, this estimation was not confirmed since no arsenate reduction was detected in a subsequent assay. This study reported for the first time the presence of phylogenetically distinct arsenate reductase genes in a Kocuria strain and evidenced the possible horizontal transfer of these genes among the endophytic bacteria.

Phylogeny and global diversity of Porodaedalea, a genus of gymnosperm pathogens in the Hymenochaetales.

Porodaedalea is a polypore genus of the Hymenochaetales that encompasses pathogens of conifer trees. In this study, we conduct a comprehensive study of the phylogeny and diversity of Porodaedalea based on collections and isolates from Europe, North America, North Africa, and Asia. Phylogenetic analysis of a two-gene data set, nuc ribosomal DNA internal transcribed spacers (ITS1-5.8S-ITS2 = ITS) and translation elongation factor 1-alpha (tef1), shows that 20 terminal clades that correspond to phylogenetic species well supported within Porodaedalea. Based on morphological evidence, five new species, P. alpicola, P. indica, P. kesiyae, P. microsperma, and P. yunnanensis, are described and illustrated. In addition, four still unnamed lineages are detected in North America and East Asia.

The Inocybe geophylla group in North America: a revision of the lilac species surrounding I. lilacina.

The Inocybe geophylla group is circumscribed based on phylogenetic analysis of DNA sequences largely sampled from North America and Europe. Twenty-nine phylogenetic species are uncovered after analysis of combined nuc 28S rDNA (28S) and RNA polymerase II second largest subunit (rpb2) DNA sequence data. Species in the I. geophylla group share the presence of a cortina, silky-fibrillose pileus and stipe, pruinose stipe apex, spermatic odor, thick-walled hymenial cystidia, and smooth amygdaliform or elliptical basidiospores. Within the group, as many as five phylogenetic species attributable to I. lilacina and allies form a strongly supported clade based on analysis of nuc ITS1-5.8S-ITS2 rDNA (ITS [internal transcribed spacer]), 28S, and rpb2 data. However, all lilac-colored species do not form a monophyletic group. Sufficient morphological and ecological data are present to document four of the I. lilacina subgroup species, two of which are described from North America as new: I. ionocephala and I. sublilacina. Inocybe lilacina is recircumscribed based on sequencing the holotype and is distributed in the eastern United States under pines and/or hardwoods. Inocybe pallidicremea is a widespread and common conifer associate in mostly northern parts of North America, to which the name I. lilacina was previously applied. Descriptions, photographs, line drawings, and a taxonomic key to lilac species in the I. lilacina subgroup from North America are provided. Well-documented collections, especially notes on gross morphology and ecology, are needed to continue to assess and describe the high taxonomic variation in the I. lilacina subgroup and its allies worldwide.

Growing Azolla to produce sustainable protein feed: the effect of differing species and CO2 concentrations on biomass productivity and chemical composition.

BACKGROUND: Since available arable land is limited and nitrogen fertilizers pollute the environment, cropping systems ought to be developed that do not rely on them. Here we investigate the rapidly growing, N2 -fixing Azolla/Nostoc symbiosis for its potential productivity and chemical composition to determine its potential as protein feed. RESULTS: In a small production system, cultures of Azolla pinnata and Azolla filiculoides were continuously harvested for over 100 days, yielding an average productivity of 90.0-97.2 kg dry weight (DW) ha-1 d-1 . Under ambient CO2 levels, N2 fixation by the fern's cyanobacterial symbionts accounted for all nitrogen in the biomass. Proteins made up 176-208 g kg-1 DW (4.9 × total nitrogen), depending on species and CO2 treatment, and contained more essential amino acids than protein from soybean. Elevated atmospheric CO2 concentrations (800 ppm) significantly boosted biomass production by 36-47%, without decreasing protein content. Choice of species and CO2 concentrations further affected the biomass content of lipids (79-100 g kg-1 DW) and (poly)phenols (21-69 g kg-1 DW). CONCLUSIONS: By continuous harvesting, high protein yields can be obtained from Azolla cultures, without the need for nitrogen fertilization. High levels of (poly)phenols likely contribute to limitations in the inclusion rate of Azolla in animal diets and need further investigation. © 2018 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The homothallic mating-type locus of the conifer needle endophyte Phialocephala scopiformis DAOMC 229536 (order Helotiales).

We describe the complete mating-type (MAT) locus for Phialocephala scopiformis Canadian Collection of Fungal Cultures (DAOMC) 229536 - a basal lineage within Vibrisseaceae. This strain is of interest due to its ability to produce the important antiinsectan rugulosin. We also provide some of the first insights into the genome structure and gene inventory of nonclavicipitalean endophytes. Sequence was obtained through shotgun sequencing of the entire P. scopiformis genome, and the MAT locus was then determined by comparing this genomic sequence to known MAT loci within the Phialocephala fortinii s.l.-Acephala applanata species complex. We also tested the relative levels of sequence conservation for MAT genes within Vibrisseaceae (n = 10), as well as within the Helotiales (n = 27). Our results: (1) show a homothallic gene arrangement for P. scopiformis [MAT1-1-1, MAT1-2-1, and MAT1-1-3 genes are present], (2) increase the genomic survey of homothallism within Vibrisseaceae, (3) confirm that P. scopiformis contains a unique S-adenosyl-l-methionine-dependent methyltransferase (SAM-Mtase) gene proximal to its MAT locus, while also lacking a cytoskeleton assembly control protein (sla2) gene, and (4) indicate that MAT1-1-1 is the more useful molecular marker amongst the MAT genes for phylogenetic reconstructions aimed at tracking evolutionary shifts in reproductive strategy and/or MAT loci gene composition within the Helotiales.

Interactions affect hyphal growth and enzyme profiles in combinations of coniferous wood-decaying fungi of Agaricomycetes.

Fomitopsis pinicola is a species of Polyporales frequently encountered in Nordic temperate and boreal forests. In nature, the fungus causes destructive brown rot in wood, colonizing tree trunks often occupied by other Basidiomycota species. We mimicked these species-species interactions by introducing F. pinicola to five white rot species, all common saprotrophs of Norway spruce. Hyphal interactions and mycelial growth in various combinations were recorded, while activities of lignocellulose-acting CAZymes and oxidoreductases were followed in co-cultures on two different carbon-source media. Of the species, Phlebia radiata and Trichaptum abietinum were the strongest producers of lignin-modifying oxidoreductases (laccase, manganese peroxidase) when evaluated alone, as well as in co-cultures, on the two different growth media (low-nitrogen liquid medium containing ground coniferous wood, and malt extract broth). F. pinicola was an outstanding producer of oxalic acid (up to 61 mM), whereas presence of P. radiata prevented acidification of the growth environment in the liquid malt-extract cultures. When enzyme profiles of the species combinations were clustered, time-dependent changes were observed on wood-supplemented medium during the eight weeks of growth. End-point acidity and production of mycelium, oxalic acid and oxidoreductase activities, in turn clustered the fungal combinations into three distinct functional groups, determined by the presence of F. pinicola and P. radiata, by principal component analysis. Our findings indicate that combinations of wood-decay fungi have dramatic dynamic effects on the production of lignocellulose-active enzymes, which may lead to divergent degradative processes of dead wood and forest litter.

Lactarius subgenus Russularia (Basidiomycota, Russulales): novel Asian species, worldwide phylogeny and evolutionary relationships.

Lactarius subg. Russularia is a large group of milkcaps occurring almost worldwide and dominant in many ecosystems. In this study we focus on new diversity, evolutionary relationships, divergence time, and origin of the subgenus. Six conifer symbionts are described as new to science: Lactarius atrii, L. aurantionitidus, L. dombangensis, L. flavigalactus, L. lachungensis, and L. sikkimensis. Species delimitation is assessed based on the concordance between morphological characteristics and an ITS phylogeny. Infrageneric relationships were studied using a phylogeny constructed from concatenated ITS-rpb2 data using Maximum Likelihood and Bayesian inference. Results show that species in this subgenus do not cluster together according to their geographic origin. Intercontinental sister relationships between Europe/Asia/North America are common but actual conspecificity is rare. This result suggests that allopatric speciation has played an important role within this subgenus. Only few morphological characteristics tend to be phylogenetically informative, with the most important being presence or absence of true cystidia and the pileipellis structure. Two datasets were generated in order to estimate the age of L. subg. Russularia. The results suggest the origin of L. subg. Russularia to be in the Mid Miocene period.

Mumia xiangluensis sp. nov., isolated from the rhizosphere of Peucedanum praeruptorum Dunn.

During an investigation of microbial diversity in medicinal herbs, a novel actinobacterium, strain NEAU-KD1(T) was isolated from the rhizosphere of Peucedanum praeruptorum Dunn collected from Xianglu Mountain in Heilongjiang Province, northeast China and characterized using a polyphasic approach. The organism was found to have the typical chemotaxonomic and morphological characteristics of the genus Mumia. Cells were observed to be non-spore-forming and irregular cocci. The cell wall was found to contain LL-diaminopimelic acid as the cell wall diamino acid. The whole-cell sugars were detected as galactose and rhamnose and the predominant menaquinone was identified as MK-9(H4). The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylglycerol, phosphoglycolipid and five unidentified phospholipids. The major cellular fatty acids were determined to be composed of C16:0, 10-methyl C18:0 and C18:1ω7c. The phylogenetic analysis based on 16S rRNA gene sequence also indicated that strain NEAU-KD1(T) belongs to the genus Mumia and with high sequence similarity to Mumia flava NBRC 109973(T) (97.6 % sequence similarity). The results of DNA-DNA hybridization and the phenotypic characteristics indicated that strain NEAU-KD1(T) could be distinguished from its close phylogenetic relative. Thus, strain NEAU-KD1(T) can be concluded to represent a novel species of the genus Mumia, for which the name Mumia xiangluensis sp. nov. is proposed. The type strain is NEAU-KD1(T) (=CGMCC 4.7305(T) = DSM 101040(T)).

Catellatospora vulcania sp. nov. and Catellatospora paridis sp. nov., two novel actinobacteria isolated from volcanic sediment and the rhizosphere of Paris polyphylla.

Two novel actinobacteria, designated strains NEAU-JM1(T) and NEAU-CL2(T), were isolated from volcanic sediment and the rhizosphere soil of Paris polyphylla, respectively, collected from Jiling and Heilongjiang Province, China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The 16S rRNA gene sequence analysis showed that the two novel isolates exhibit 99.5 % 16S rRNA gene sequence similarity with each other. The phylogenetic analysis based on 16S rRNA gene sequence of strain NEAU-JM1(T) showed it to be closely related to Catellatospora methionotrophica JCM 7543(T) (99.4 % sequence similarity), Catellatospora coxensis DSM 44901(T) (99.4 %), Catellatospora citrea DSM 44097(T) (99.3 %) and Catellatospora chokoriensis JCM 12950(T) (99.2 %), and that of strain NEAU-CL2(T) to C. citrea DSM 44097(T) (99.4 %), C. methionotrophica JCM 7543(T) (99.3 %), C. chokoriensis JCM 12950(T) (99.3 %) and C. coxensis DSM 44901(T) (99.2 %). However, the DNA-DNA hybridization value between strains NEAU-JM1(T) and NEAU-CL2(T) was 62.2 %, and the values between the two strains and their close phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their close phylogenetic relatives. Thus, strains NEAU-JM1(T) and NEAU-CL2(T) can be concluded to represent two novel species of the genus Catellatospora, for which the names Catellatospora vulcania sp. nov. and Catellatospora paridis sp. nov. are proposed. The type strains are NEAU-JM1(T) (=CGMCC 4.7174(T) = JCM 30054(T)) and NEAU-CL2(T) (=CGMCC 4.7236(T) = DSM 100519(T)), respectively.

Bioactive Secondary Metabolites Produced by the Fungal Endophytes of Conifers.

This is a review of bioactive secondary metabolites isolated from conifer-associated endophytic fungi from 1990-2014. This includes compounds with antimicrobial, anti-inflammatory, anti-proliferative and cytotoxic activity towards human cancer cell lines, and activity against either plant pathogens or plant insect pests. Compounds that were originally reported without associated activity were included if other studies ascribed activity to these compounds. Compounds were not included if they were exclusively phytotoxic or if they were isolated from active extracts but were not determined to be the active component of that extract.

Microbispora sp. LGMB259 endophytic actinomycete isolated from Vochysia divergens (Pantanal, Brazil) producing ß-carbolines and indoles with biological activity.

Endophytic actinomycetes encompass bacterial groups that are well known for the production of a diverse range of secondary metabolites. Vochysia divergens is a medicinal plant, common in the "Pantanal" region (Brazil) and was focus of many investigations, but never regarding its community of endophytic symbionts. During a screening program, an endophytic strain isolated from the V. divergens, was investigated for its potential to show biological activity. The strain was characterized as Microbispora sp. LGMB259 by spore morphology and molecular analyze using nucleotide sequence of the 16S rRNA gene. Strain LGMB259 was cultivated in R5A medium producing metabolites with significant antibacterial activity. The strain produced 4 chemically related ß-carbolines, and 3 Indoles. Compound 1-vinyl-ß-carboline-3-carboxylic acid displayed potent activity against the Gram-positive bacterial strains Micrococcus luteus NRRL B-2618 and Kocuria rosea B-1106, and was highly active against two human cancer cell lines, namely the prostate cancer cell line PC3 and the non-small-cell lung carcinoma cell line A549, with IC50 values of 9.45 and 24.67 µM, respectively. 1-Vinyl-ß-carboline-3-carboxylic acid also showed moderate activity against the yeast Saccharomyces cerevisiae ATCC204508, as well as the phytopathogenic fungi Phyllosticta citricarpa LGMB06 and Colletotrichum gloeosporioides FDC83.

Relación in vitro entre cylindrocarpon cylindroides y neonectria fuckeliana asociados a cancros fustales de pinus radiata en Chile / In vitro relationship between cylindrocarpon cylindroides and neonectria fuckeliana associated with stem cankers on pinus radiata in Chile

Desde el punto de vista estrictamente micológico, se ha discutido por décadas la relación de Cylindrocarpon cylindroides como anamorfo de Neonectria fuckeliana, lo que entre otras consecuencias, ha incidido en la controversial ubicación taxonómica de este teleomorfo. Se añade a esto que en patología forestal se ha brindado poca atención a la presencia de C. cylindroides dentro del ciclo de la enfermedad causada por Neonectria fuckeliana en coníferas, razón por la cual los estudios específicos respecto de este anamorfo han sido también muy escasos. El presente trabajo tuvo como objetivo verificar la relación entre C. cylindroides y Neonectria fuckeliana, asociados a cancros en troncos de Pinus radiata en Chile, mediante el análisis de las características macro y micromorfológicas que ellos manifiestan en cultivos in vitro, además de análisis de tipo molecular. Se pudo constatar que ambos agentes dieron origen a colonias con idénticas características macro y micromorfológicas, destacando entre éstas la producción de abundantes conodióforos y microconidios del tipo Acremonium, el cual ya ha sido señalado en anteriores estudios como estado anamorfo de Neonectria fuckeliana. El que este mismo estado asexual del tipo Acremonium haya sido la manifestación in vitro tanto de C. cylindroides como de Neonectria fuckeliana, permitió confirmar la relación existente entre estos dos agentes en estudio, la que también fue ratificada mediante los análisis moleculares realizados.

From the point of view strictly mycological has been discussed for decades the relationship of Cylindrocarpon cylindroides as anamorph of Neonectria fuckeliana, which among other consequences, has influenced the controversial taxonomic status of this teleomorph. Add to this that in forest pathology has given little attention to the presence of C. cylindroides within the cycle of the disease caused by Neonectria fuckeliana on conifers, why specific studies regarding this anamorph have been very few. This study aimed to verify the relationship between C. cylindroides and Neonectria fuckeliana associated with cankers on trunks of Pinus radiata in Chile, by analyzing the macro and micromorphological characteristics they manifest in cultures in vitro, further analysis type molecular. It was found that both agents gave rise to colonies with identical macro and micromorphological characteristics, highlighting among them the production of abundant microconidia and conidiophores of Acremonium type, which has already been noted in previous studies as anamorph state of Neonectria fuckeliana. The asexual state that this same type of Acremonium has been the in vitro demons-tration of both C. cylindroides as Neonectria fuckeliana, allowed us to confirm the relationship between these two agents in the study, which was also ratified by the molecular analyzes performed.

Nocardia bhagyanesis sp. nov., a novel actinomycete isolated from the rhizosphere of Callistemon citrinus (Curtis), India.

A novel actinomycete strain, designated VRC07(T), was isolated from a Callistemon citrinus rhizosphere sample collected from Hyderabad, India. Its taxonomic status was determined by using polyphasic approach. It is a Gram-positive, aerobic, non-motile, weakly acid-fast strain. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain VRC07(T) is a member of the genus Nocardia. The highest levels of 16S rRNA gene sequence similarity was found between the strains Nocardia niwae W9241(T) (99.6 %), Nocardia amikacinitolerans W9988(T) (99.3 %) and Nocardia arthritidis IFM 10035(T) (98.9 %); similarity to other type strains of the genus Nocardia was below 98.7 %. The organism had chemical and morphological features consistent with its classification in the genus Nocardia such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan. Arabinose and galactose as the diagnostic sugars. Diagnostic polar lipids were phosphatidylinositol, diphosphatidylglycerol, and phosphatidylglycerol. The predominant menaquinone was MK-8(H4, ω-cycl). The major fatty acids were C16:0, C18:0, C18:1 w9c, C18:0 10-methyl TBSA and sum in feature 3 (16:1 w7c/16:1 w6c). The G+C content of the genomic DNA was 68.5 mol%. The DNA-DNA relatedness data, together with phenotypic differences clearly distinguished the isolate from its closest relatives. On the basis of these phenotypic and genotypic data, the isolate represents a novel species, for which the name Nocardia bhagyanesis sp. nov., is proposed. The type strain is VRC07(T) (=KCTC 29209(T) = MTCC 11725(T) = ATCC BAA-2548).

Expression analysis of the impact of culture filtrates from the biocontrol agent, Phlebiopsis gigantea on the conifer pathogen, Heterobasidion annosum s.s. Transcriptome.

Phlebiopsis gigantea has been routinely used as the biological control agent for the conifer pathogen Heterobasidion annosum sensu lato, but the actual mechanism for the biocontrol process is not known. To investigate the effect of secreted molecules from culture filtrate produced by P. gigantea on the gene expression profile of H. annosum s.s., microarray analysis was used. Analysis of the differentially expressed genes led to the identification of genes with diverse functions. A major proportion of the up- and downregulated genes were either uncharacterized or genes whose functions were not known. A number of genes coding for proteins involved in metabolism, transport, and signal transduction were differentially downregulated; comparatively lower number of such genes were upregulated. Some genes involved in transport (polyamine transporters, 2573-fold, P = 0.002) and metabolism (endoglucanase, 622.5-fold, P = 0.002, cytochrome P450, 133.2-fold, P = 0.05) showed high transcript fold changes and were statistically significantly upregulated. Genes encoding defense-related proteins such as hydrophobins were either downregulated or expressed at relatively low levels. Further analysis of the effect of the culture filtrate on glucose metabolism showed downregulation of some key enzymes at the early stage of the glycolytic pathway while some genes were upregulated at the later stage of the pathway. A subset of the genes were selected and used to validate the micro-array result by quantitative real time polymerase chain reaction (qPCR) method. Generally, the high transcript levels of genes encoding several biochemically important genes (protein kinases, major facilitator superfamily polyamine transporters, endoglucanase, cytochrome P450, endoglucanase) suggests their potential functional relevance in signal perception, stress tolerance, cell defenses, and detoxification of toxic molecules during competitive interaction. These results have provided further insights into possible molecular and genetic factors underlying the response of H. annosum to metabolites from P. gigantea during interspecific interaction.

Cortinarius sanguineus and equally red species in Europe with an emphasis on northern European material.

The red species of Cortinarius subgenus Dermocybe in Europe were studied based on morphological and molecular data. Three completely red species were recognized: C. sanguineus (syn. C. sanguineus var. aurantiovaginatus), C. puniceus (syn. C. cruentus, C. rubrosanguineus) and C. vitiosus comb. nov. Cortinarius sanguineus has dusky red to red pileus, reddish yellow mycelium and lacking or with only slightly encrusted hyphae in pileipellis. It occurs in mesic to damp forests with Picea, often on rich soil in the boreal and montane areas of Europe, presumably also in eastern Canada. Cortinarius puniceus differs from C. sanguineus by its stronger purplish red, narrower spores and spot-like encrusted hyphae in pileipellis. It grows with deciduous trees in the temperate zone of Europe. Cortinarius vitiosus is known only from Fennoscandia and occurs in dry to mesic coniferous forests. It has fairly thin, often zonate, dark red to dark reddish brown pileus, pale red mycelium, small spores and encrusted lamellar trama and pileipellis hyphae. In addition to these three species C. fervidus and C. phoeniceus occasionally have red basidiomes. The relationships of the species were inferred by analysis of ITS sequences. Our study suggests that the section Sanguinei, as earlier defined, is polyphyletic. Here the section is limited to include C. sanguineus, C. puniceus and North American D. sierraensis. The relationships with other red species were not determined. Section Dermocybe, including C. cinnamomeus, C. croceus and C. uliginosus, formed a monophyletic group, and the section Malicoriae had some support. A total of 34 new sequences are published including nine from type specimens.

Molecular phylogenetics of the Gloeophyllales and relative ages of clades of Agaricomycotina producing a brown rot.

The Gloeophyllales is a recently described order of Agaricomycotina containing a morphologically diverse array of polypores (Gloeophyllum), agarics (Neolentinus, Heliocybe) and resupinate fungi (Veluti-Veluticeps, Boreostereum, Chaetodermella), most of which have been demonstrated to produce a brown-rot mode of wood decay and are found preferentially on coniferous substrates. Multiple phylogenetic studies have included taxa of Gloeophyllales, but none have sampled the order thoroughly, and so far only ribosomal RNA genes have been used. Consequently the limits and higher level placement of the Gloeophyllales are obscure. We obtained sequence data for three protein-coding genes (rpb2, atp6, tef1) and three rRNA regions (nuc-ssu, nuc-lsu, 5.8S) in 19 species of Gloeophyllales representing seven genera and analyzed them together with a diverse set of Agaricomycotina, emphasizing Polyporales. Boreostereum, which is suspected to produce a white rot, is the sister group of the rest of the Gloeophyllales, all of which produce a brown rot. Gloeophyllum contains at least two independent clades, one of which might correspond to the genus Osmoporus. White rot and resupinate fruiting bodies appear to be plesiomorphic in Gloeophyllales. Relaxed molecular clock analyses suggest that the Gloeophyllales arose in the Cretaceous, after the origin of Pinaceae.

Arbuscular mycorrhizal infection in two morphological root types of Araucaria araucana (Molina) K. Koch / Infección por micorrizas arbusculares en dos tipos de raíces de Araucaria araucana (Molina) K. Koch

Araucaria araucana (Molina) K. Koch is a conifer distributed in the Andean-Patagonian forests in the south of Argentina and Chile. The main objective of this work was to relate the different root classes appearing in A. araucana to mycorrhizal behavior. Samples were collected in three different sites in the Lanín National Park (NW Patagonia, Argentina). Two different root classes were present in A. araucana: longitudinal fine roots (LFR) and globular short roots (GSR). Both had extensive mycorrhizal arbuscular symbiosis (AM) and presented abundant hyphae and coils in root cells, a characteristic of the anatomical Paris-type. Dark septate fungal endophytes were also observed. Values of total AM colonization were high, with similar partial AM% values for each root class. Seasonal differences were found for total and partial colonization, with higher values in spring compared to autumn. Regarding the percentage of fungal structures between root classes, values were similar for vesicles and arbuscules, but higher coil percentages were observed in GSR compared to LFR. The percentages of vesicles increased in autumn, whereas the arbuscule percentages increased in spring, coinciding with the plant growth peak. Results show that both root classes of A. araucana in Andean-Patagonian forests are associated with AM fungi, which may have ecological relevance in terms of the importance of this symbiosis, in response to soil nutrient-deficiencies, especially high P-retention.

La conífera Araucaria araucana (Molina) K. Koch se encuentra distribuida en los bosques Andino-Patagónicos de Argentina y Chile. En este trabajo se relacionaron las diferentes clases morfológicas de raíces presentes en la especie con el comportamiento micorrícico. Las muestras fueron tomadas en tres sitios del Parque Nacional Lanín (NO de Patagonia, Argentina). Se observaron dos clases diferentes de raíces: raíces finas longitudinales (RFL) y raíces cortas globulares (RCG). Ambas clases presentaron una importante simbiosis micorrícico-arbuscular (MA), con abundante desarrollo de hifas intracelulares y rulos, característico del tipo anatómico Paris. También se observaron hifas marrones septadas. Los valores totales de colonización MA fueron altos, con porcentajes parciales similares en cada clase radicular. Se encontraron diferencias estacionales en los porcentajes de infección (totales y parciales), con valores mayores en primavera comparados con los de otoño. Los porcentajes de estructuras fúngicas presentes en cada clase radicular fueron similares para vesículas y arbúsculos, pero se observaron mayores valores de rulos en las RCG que en las RFL. Los porcentajes de vesículas aumentaron en otoño, mientras que los de arbúsculos lo hicieron en primavera, en coincidencia con el pico de crecimiento vegetativo de los árboles. Los resultados muestran una fuerte asociación simbiótica MA con A. araucana y su relevancia en términos ecológicos en respuesta a la deficiencia de nutrientes en el suelo, en especial, a la alta retención de fósforo.

The genus Laetiporus (Basidiomycota, Polyporales) in East Asia.

Relationships among East Asian, North American and European Laetiporus sulphureus s. lat., a cosmopolitan brown rot species complex, were assessed with phylogenetic analyses and incompatibility tests. Three East Asian taxa, Laetiporus cremeiporus sp. nov., Laetiporus montanus and Laetiporus versisporus, are described and illustrated as well as compared with related taxa from Southeast Asia, North America and Europe. Phylogenetic analyses showed that L. cremeiporus and L. versisporus are clearly distinct species among Laetiporus taxa. The three conifer inhabiting species, Laetiporus conifericola, Laetiporus huroniensis and L. montanus, are closely related to each other. The European population of L. montanus exhibits two sequence variants of the EF1alpha: one is the same as observed in L. sulphureus in Europe and the other is that observed in East Asian population of L. montanus. A key to the known species of Laetiporus in the northern hemisphere is provided.