ITS rDNA Sequence Analysis Research Articles

Identification of pathogen of Aquilaria sinensis dieback disease

The research focused on identification the pathogen of Aquilaria sinensis dieback disease and confirmation the function of inducing agarwood formation by fungi. The morphological observation, rDNA ITS sequence analyses, Koch's postulates and pathogenicity test were used to identify the isolates. The isolates of the causal agent was Lasiodiplodia theobromae, which induced the plant to produce highly valuable agarwood. The first report of dieback disease of Aquilaria sinensis caused by L. theobromae, and our research results laid a theoretical foundation for agarwood production by using fungus.

Isolation and identification of pathogen of seedling blight of Platycodon grandiflorum

To isolate and identify pathogen of the seedling blight occurred in Platycodon grandiflorum. The morphological observation, rDNA ITS sequence analysis, and Koch's postulates were used to identify the isolates of the causal agent. The isolates of the causal agent was Rhizoctonia solani. The result confirmed that R. solani is the pathogen of seedling blight of P. grandiflorum.

First Report of Colletotrichum acutatum Associated with Stem Blight of Blueberry Plants in China.

An anthracnose disease was observed on stems of high-bush blueberry plants (Vaccinium corymbosum L.) in Liaoning Province, China in 2012. The typical symptoms consist of sudden wilting and dieback of stems during the growing season. Dark brown lesions originate from infected buds and kill portions of the stems. Lesions have grayish white centers, with the necrotic areas becoming 6 to 8 cm in length. Disinfected stem pieces were placed on potato dextrose agar (PDA) and incubated at 28°C for 5 to 7 days, after which the emerging colonies were transferred to fresh PDA. All isolates initially produced white growth, but turned pink after 7 days before becoming blackish green. The average colony diameter was 65.5 to 75.0 mm after 7 days. Conidia were aseptate, hyaline, fusiform to ellipsoid, 8.5 to 16.5 × 2.5 to 4.0 μm in size and single celled with two to seven oil globules. Setae were not found on the acervuli. These characteristics matched published descriptions of Colletotrichum acutatum (1) (teleomorph Glomerella acutata). Pathogenicity test was confirmed in 15 2-year-old healthy potted plants of cv. Berkeley. Stems of 10 plants were punctured with flamed needles and sprayed with 5 ml of conidial suspension (106 conidia per ml in sterile distilled water) of isolate LNSW1. Five control plants were inoculated with sterile distilled water. Seven days after inoculation, eight of the 10 blueberry plants exhibited stem lesions, leaf chlorosis, followed by branch dieback 15 days post-inoculation. The symptoms were similar to those observed on diseased plants in the field, and no lesions were observed on control plants. The pathogen was reisolated from the margin of lesions and identified by colony growth characteristics on PDA. PCR amplification of one isolate (LNSW1) was carried out by utilizing the universal rDNA-ITS primer pair ITS1/ITS4. The sequence (557 bp) of isolate LNSW1 (GenBank Accession No. JX392857) showed 99% identity to G. acutata (AB443950) and C. acutatum (AJ749672) in a BLAST search. An approximately 490-bp fragment was amplified from LNSW1 by the species-specific primer pair CaInt2/ITS4 (2). The pathogen was identified as G. acutata (asexual stage: C. acutatum J.H. Simmonds) on the basis of morphological characters, rDNA-ITS sequence analysis, and a PCR product with species-specific primers. To our knowledge, this is the first report of C. acutatum in high-bush blueberry plants in China.

Diversity analysis of Beauveria bassiana isolated from infected silkworm in southwest China based on molecular data and morphological features of colony

Beauveria bassiana is an important entomopathogenic fungus that not only often causes infection and epidemics of wild insects but some strains also show pathogenicity to the silkworm, Bombyx mori. The present study is about diversity of B. bassiana isolated from the silkworm in southwest China. Five strains of B. bassiana were isolated from infected silkworm. Two isolates, GXtr1009 and GXtr1010, were isolated from infected silkworms treated with two kinds of biological pesticides applied in Guangxi province, and three isolates, SCsk1006, YNsk1106 and GXsk1011, were collected from naturally infected silkworms from different geographical locations in Yunnan and Sichuan. All of the isolates showed highly similar conidia and conidial fructification, but the colony characteristics demonstrated great differences among the isolates. The ITS and 18S rDNA sequence analysis was sufficient to identify all five isolates as B. bassiana. However, the dendrogram, based on the ISSR data, produced two large genetic groups. GXtr1009 and GXtr1010 comprised one group, and SCsk1006, YNsk1106 and GXsk1011 converged in a different large group. The results suggested that, although all of these five B. bassiana strains were pathogenic to silkworms, strains of biological pesticides could be differentiated from strains of naturally infected silkworm via ISSR analysis.

First Report of Leaf Spot Disease Caused by Colletotrichum gloeosporioides on Chinese Bean Tree in China.

Chinese bean tree, Catalpa fargesii f. duciouxii (Dode) Gilmour, is an ornamental arbor plant. Its roots, leaves, and flowers have long been used for medicinal purposes in China. During July 2010, severe outbreaks of leaf spot disease on this plant occurred in Kunming, Yunnan Province. The disease incidence was greater than 90%. The symptoms on leaves began as dark brown lesions surrounded by chlorotic halos, and later became larger, round or irregular spots with gray to off-white centers surrounded by dark brown margins. Leaf tissues (3 × 3 mm), cut from the margins of lesions, were surface disinfected in 0.1% HgCl2 solution for 3 min, rinsed three times in sterile water, plated on potato dextrose agar (PDA), and incubated at 28°C. The same fungus was consistently isolated from the diseased leaves. Colonies of white-to-dark gray mycelia formed on PDA, and were slightly brown on the underside of the colony. The hyphae were achromatic, branching, septate, and 4.59 (±1.38) μm in diameter on average. Perithecia were brown to black, globose in shape, and 275.9 to 379.3 × 245.3 to 344.8 μm. Asci that formed after 3 to 4 weeks in culture were eight-spored, clavate to cylindrical. The ascospores were fusiform, slightly curved, unicellular and hyaline, and 13.05 to 24.03 × 10.68 to 16.02 μm. PCR amplification was carried out by utilizing universal rDNA-ITS primer pair ITS4/ITS5 (2). Sequencing of the PCR products of DQ1 (GenBank Accession No. JN165746) revealed 99% similarity (100% coverage) with Colletotrichum gloeosporioides isolates (GenBank Accession No. FJ456938.1, No. EU326190.1, No. DQ682572.1, and No. AY423474.1). Phylogenetic analyses (MEGA 4.1) using the neighbor-joining (NJ) algorithm placed the isolate in a well-supported cluster (>90% bootstrap value based on 1,000 replicates) with other C. gloeosporioides isolates. The pathogen was identified as C. gloeosporioides (Penz.) Penz. & Sacc. (teleomorph Glomerella cingulata (Stoneman) Spauld & H. Schrenk) based on the morphological characteristics and rDNA-ITS sequence analysis (1). To confirm pathogenicity, Koch's postulates were performed on detached leaves of C. fargesii f. duciouxii, inoculated with a solution of 1.0 × 106 conidia per ml. Symptoms similar to the original ones started to appear after 10 days, while untreated leaves remained healthy. The inoculation assay used three leaves for untreated and six leaves for treated. The experiments were repeated once. C. gloeosporioides was consistently reisolated from the diseased tissue. C. gloeosporioides is distributed worldwide causing anthracnose on a wide variety of plants (3). To the best of our knowledge, this is the first report of C. gloeosporioides causing leaf spots on C. fargesii f. duciouxii in China.

Pseudoidium javanicum, a new species of powdery mildew on Acalypha spp. from Indonesia

Pseudoidium javanicum is proposed as a new species based on analyses of 28S, ITS and IGS rDNA sequences, and morphological data. This new species was found on Acalypha wilkesiana var. marginata, A. argentea, and A. cristata collected from Cibodas Botanical Garden, Bogor (West Java Province, Indonesia). Our analyses showed that all these specimens have identical rDNA sequences and similar morphological characteristics. They form a distinct clade separated from other species of Erysiphaceae. Pseudoidium javanicum differs from Erysiphe acalyphae by having shorter conidiophores and foot cells 1–3 times as long as the 0–2 following cells. The conidial size of Ps. javanicum is also smaller than that of E. jatrophae.

Rhizoctonia solani AG 2-2IIIB에 의한 마 뿌리썩음병의 한국 내 발생

2011년 안동과 진주의 마 재배포장에서 줄기 지제부 및 괴근썩음 증상이 나타났다. 병징을 나타내는 부위로부터 Rhizoctonia와 유사 속에 속하는 20개 균주가 분리되었다. rDNA-internal transcribed spacer(ITS) 염기서열 상동성을 기초로 8균주는 Rhizoctonia solani, 12균주는 Ceratobasidium sp.로 동정되었다. rDNA-ITS 염기서열의 cluster 분석에 의해 R. solani에 속하는 8개 균주 중 7개 균주는 균사융합군 AG 2-2IIIB, 1균주는 AG 1-1A에 속하였다. 또한, Ceratobasidium sp.에 속하는 12균주 중 7균주는 AG-Fa, 3균주는 AG-A, 나머지 2균주는 각각 AG-Fb와 AG-O에 속하였다. R. solani AG 2-2IIIB 균주들은 마의 줄기와 괴근에 병원성이었으나 R. solani AG 1-1A와 모든 Ceratobasidium sp. 균주는 비병원성이라는 것이 확인되었다. 이 결과는 조사지역에서 R. solani AG 2-2IIIB가 마의 줄기 및 괴근썩음병을 일으키는 중요한 병원균이라는 것을 나타낸다. 이 연구는 국내에서 R. solani AG 2-2IIIB에 의한 마 뿌리썩음병에 대하여 처음으로 보고하는 것이다. Stem canker and tuber rot symptoms were observed on yam grown in Andong and Jinju, Korea in 2011. A total of 20 isolates of Rhizoctonia and allied fungi were obtained from the symptomatic plants. Among the isolates, 8 isolates were identified as Rhizoctonia solani and 12 isolates as Ceratobasidium sp. based on rDNA-internal transcribed spacer (ITS) sequence similarity. In the cluster analysis of rDNA-ITS sequences, 7 isolates of R. solani belonged to AG 2-2IIIB and remaining one to AG 1-1A. In addition, among the 12 isolates of Ceratobasidium sp., 7 isolates belonged to AG-Fa, three isolates to AG-A and the other two isolates to AG-Fb and AG-O, respectively. Pathogenicity tests showed that all the R. solani AG 2-2IIIB isolates are pathogenic on stem and tuber of yam but R. solani AG 1-1A and all the Ceratobasidium isolates are non-pathogenic. The results indicate that R. solani AG 2-2IIIB is an important pathogen causing stem canker and tuber rot on yams grown in the study areas. This is the first report of R. solani AG 2-2IIIB causing stem canker and tuber rot of yam in Korea.

Cladosporium cladosporioides XJ-AC03, an aconitine-producing endophytic fungus isolated from Aconitum leucostomum

The endophytic fungus XJ-AC03, which was isolated from the healthy roots of Aconitum leucostomum, produced aconitine when grown in potato dextrose agar (PDA) medium. The presence of aconitine was confirmed by the chromatographic and spectroscopic analyses. The yield of aconitine was recorded as 236.4 μg/g by high performance liquid chromatography (HPLC). The mass spectrometry was shown to be identical to authentic aconitine. Further analysis with nuclear magnetic resonance (NMR) spectroscopy to show the chemical structure of the fungal aconitine indicated that the fungal aconitine produced an NMR spectrum identical to that of authentic aconitine. Strain XJ-AC03 was identified as Cladosporium cladosporioides by its characteristic culture morphology and ITS rDNA sequence analysis.

Two new resupinate Thelephorales (Basidiomycota, Agaricomycetes) from Guinea (West Africa)

Tomentella minispora and T. baroensis are described as new species from West Africa based on anatomical evidence, ITS rDNA sequence analyses and molecular-phylogenetic inference. T. minispora is characterized by subglobose to globose small basidiospores with size ranging from 5.5 μm to 7.5 μm, short aculei of up to 0.8 μm and strongly inflated subhymenial hyphae (up to 23 μm). Both, sequence analyses and molecular-phylogenetic inference indicate the proximity of T. minispora to the clampless species T. amyloapiculata and T. fuscocinerea. Tomentella baroensis is characterized by the presence of extremely large young basidia (up to 20 μm wide), ellipsoid to lobed basidiospores of 8–13 μm and short aculei not exceeding 1 μm long. So far such large young basidia are known only from T. terrestris. However, ITS rDNA sequence analyses revealed that T. baroensis deviates from T. terrestris by at least 17 to 18%. Sequence analyses suggest T. ellisii as the most similar species to T. baroensis, whilst molecular-phylogenetic inference places it close to T. pisoniae and T. hjortstamiana, together with T. ellisii originating from deeper branch of the clade. Detailed anatomy and original iconographs are provided for both new species. Anatomical comparisons are made between the newly described species with their anatomically and molecularphylogenetically close ones.

Endophytic fungi harbored in Cannabis sativa L.: diversity and potential as biocontrol agents against host plant-specific phytopathogens

The objective of the present work was isolation, phylogenetic characterization, and assessment of biocontrol potential of endophytic fungi harbored in various tissues (leaves, twigs, and apical and lateral buds) of the medicinal plant, Cannabis sativa L. A total of 30 different fungal endophytes were isolated from all the plant tissues which were authenticated by molecular identification based on rDNA ITS sequence analysis (ITS1, 5.8S and ITS2 regions). The Menhinick’s index revealed that the buds were immensely rich in fungal species, and Camargo’s index showed the highest tissue-specific fungal dominance for the twigs. The most dominant species was Penicillium copticola that could be isolated from the twigs, leaves, and apical and lateral buds. A detailed calculation of Fisher’s log series index, Shannon diversity index, Simpson’s index, Simpson’s diversity index, and Margalef’s richness revealed moderate overall biodiversity of C. sativa endophytes distributed among its tissues. The fungal endophytes were challenged by two host phytopathogens, Botrytis cinerea and Trichothecium roseum, devising a dual culture antagonistic assay on five different media. We observed 11 distinct types of pathogen inhibition encompassing a variable degree of antagonism (%) on changing the media. This revealed the potential chemodiversity of the isolated fungal endophytes not only as promising resources of biocontrol agents against the known and emerging phytopathogens of Cannabis plants, but also as sustainable resources of biologically active and defensive secondary metabolites.

Antimicrobial activity and molecular characterization of an endophytic fungus, Quambalaria sp. isolated from Ipomoea carnea

An endophytic fungus displaying considerable antimicrobial activity was isolated from stem tissue of an invasive plant species, Ipomoea carnea. The fungus was identified as Quambalaria sp. and confirmed by ITS rDNA sequence analysis. A BLAST search result of the sequence indicated 97 % homology with Quambalaria cyanescens. Crude metabolites of the fungus showed considerable antimicrobial activity against a panel of clinically significant microorganisms. The metabolites showed highest in vitro activity against Shigella dysenteriae followed by Escherichia coli and Candida albicans. Optimum metabolites production required neutral pH and a 15-day incubation period. Bark extracts amended with fungal media demonstrated higher antimicrobial activity. Optimum metabolites activity was recorded in Czapek Dox broth amended with leaf extracts (CDB + LE) of the host plant. The metabolites showed UV λ-max in ethyl acetate at 284.6 nm with an absorbance value of 1.093. Phylogenetic tree generated by the Maximum Parsimony method showed clustering of our isolate with Q. cyanescens with supported bootstrap of 65 %. Species of Quambalaria are pathogens to Eucalyptus and occurrence of this fungus as endophytes support it to be a latent pathogen. Sequence base analysis and RNA secondary structure study also confirmed such a relationship. Secondary structural features like two hinges and a 5’ dangling end were found to be unique to our isolate. These structural features can also be used as potential barcodes for this fungus. The findings indicate that invasive plant species can be a reliable source of novel endophytes with rich antimicrobial metabolites. The study also validates the assumption that endophytes can become parasites and share a close affinity.

Antrodiella chinensis sp. nov., a Chinese representative of the Antrodiella americana complex

A new wood-decaying polypore, Antrodiella chinensis, is described and illustrated from China based on morphological characteristics and phylogenetic analysis of rDNA ITS sequences. The new species belongs to the Antrodiella americana complex, and is characterized by an annual habit, resupinate basidiocarps, cream to straw coloured pore surface with larger and regular pores, and oblong-ellipsoid basidiospores. Discriminating characteristics between the new species and the closely related species in the complex are discussed.

Generic and species concepts in Microglena (previously the Chlamydomonas monadina group) revised using an integrative approach†

Traditionally the genus Microglena Ehrenberg has been used to contain species that belong to the Chrysophyceae; however, the type species of Microglena, M. monadina, represents a green alga, which was later transferred to the genus Chlamydomonas. The taxonomic status of the genus has therefore remained unclear. We investigated 15 strains previously assigned to C. monadina and two marine species (C. reginae and C. uva-maris) using an integrative approach. Phylogenetic analyses of SSU and ITS rDNA sequences revealed that all strains form a monophyletic lineage within the Chlorophyceae containing species from different habitats. The strains studied showed similar morphology with respect to cell shape and size, but showed differences in chloroplast and pyrenoid structures. Some representatives of this group have the same type of sexual reproduction (homothallic advanced anisogamy). Three different morphotypes could be recognized. Strains belonging to type I have a cup-shaped chloroplast with a massive basal part, in which a large, single, ellipsoidal pyrenoid is located. The members of type II also have a cup-shaped chloroplast, which is partly lobed and has a thinner basal part than type I; here the pyrenoid is half-ring or horseshoe-shaped and occupies different positions in the chloroplast depending on the strain. The strains of type III have multiple pyrenoids, which appear to have developed from the subdivision of a single ring-shaped pyrenoid into several parts. We compared the results of our morphological investigations with the literature and found that 15 strains could be identified with existing species. Two strains did not fit with any described species. As a result of our study, we transfer all strains to the genus Microglena, propose 11 new combinations, and describe two new species. Comparison of the ITS-1 and ITS-2 secondary structures confirmed the species delineations. All species have characteristic compensatory base changes in their ITS secondary structures and are supported by ITS-2 DNA barcodes. †Molecular phylogeny and taxonomic revision of Chlamydomonas (Chlorophyta). II.

Pseudochaete latesetosa and P. subrigidula spp. nov. (Hymenochaetales, Basidiomycota) from China based on morphological and molecular characters

Pseudochaete latesetosa and P. subrigidula collected from southern China are described as new species based on a combination of morphological and molecular characters. Both the species have narrowly cylindrical to allantoid basidiospores and encrusted setae, which are diagnostic characters of Pseudochaete. P. latesetosa is similar to P. tabacina, except that the latter has resupinate, effused-reflexed or pileate basidiocarps and well-developed cortex and tomentum. P. subrigidula resembles P. rigidula in macroscopical and anatomical structures, but the latter has shorter setae and basidiospores. The phylogenetic results inferred from analysis of combined ITS and nLSU rDNA sequences show that the two species were nested within the Pseudochaete clade and were separated from other sampled species as well as from each other. Illustrated descriptions of these two species are provided.

Morphological and molecular identification of pathogenic fungal of post-harvest tomato fruit during storage

Three pathogenic fungi of tomato were separated by using transconjugate method and identified by morphological characteristics and internal transcribed spacers ribosomal DNA (ITS rDNA) sequence analysis. Morphological studies showed that FQ1, FQ2 and FQ3 generated spores on potato dextrose agar (PDA) plates and slides. The ITS rDNA regions of these pathogens were amplified by polymerase chain reaction (PCR) and then sequenced. After analyses of their ITS rDNA sequences, a phylogenetic tree was obtained from the program Neighbor-joining (NJ) together with the sequence of related strains which were downloaded from GenBank. The results of morphological observation and molecular detection of three fungi showed that FQ1 belong to Alternaria alternate. FQ2 belong toCladosporium. FQ3 belongs to Fusarium sp. Key words: Tomato fruit, postharvest diseases, fungi, ITS rDNA sequence analysis, identification.

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.

Identification and secondary metabolites of endophytic fungal strain PR35 from Paeonia delavayi

To identify the endophytic fungal strain PR35 separated from Paeonia delavayi and study chemical constituents of its secondary metabolites. The fungal strain PR35 was identified by morphological observation and ITS rDNA sequence analysis. Various chromatographic methods were adopted to separate and purify its secondary metabolites, and their structures were identified by physiochemical properties and spectral data The fungal strain PR35 was identified as Trichoderma longibrachiatum. Five compounds were separated from fermentation products of fungal strain PR35 and identified as 1-(2,6-dihydroxyphenyl)-3-hydroxybutan-1-one (1), 1-(2,6-dihydroxypheny) propan-1-one (2), 1-(2,4,6-trihydroxyphenyl) butan-1-one (3), 4-methoxy-1-naphthol (4), and cerevisterol (5). Among them, compounds 1-3 showed notable antifungal activities against Botrytis cinerea, Fusarium avenaceum and Hormodendrum compactum. The endophytic fungus T. longibrachiatum was separated from the plant P. delavayi for the first time. Five compounds were first separated from endophytic fungus of P. delavayi. Among them, compound 4 was separated from microbial fermentation products for the first time.

Sclerotium rolfsii에 의한 수세미오이 흰비단병

경상남도농업기술원 시험포장에서 S. rolfsii에 의한 수세미오이 흰비단병 증상이 발생하였다. 병징은 과실이 수침상으로 무르고 갈색으로 변하면서 썩는다. 병반부와 토양 표면에 흰색의 곰팡이가 발생하며 갈색의 작은 둥근 균핵이 형성되었다. 감자한천배지에서 균총은 흰색이고 잘 자라며 배양기간이 경과됨에 따라 갈색의 작은 둥근균핵을 많이 형성하였다. 균핵의 크기는 1-3 mm이며 균사의 폭은 4-9 <TEX>${\mu}m$</TEX>였다. 균사생육과 균핵 형성 적온은 <TEX>$30^{\circ}C$</TEX>이었다. 균사특유의 clamp connection이 관찰되었다. 수세미오이에서 발생한 병징과 병원균의 균학적 특징, 그리고 ITS rDNA 염기서열 비교분석 결과, 이 병을 Sclerotium rolfsii Saccardo에 의한 수세미오이 흰비단병으로 명명하고자 제안한다. Sclerotium rot of sponge gourd occurred at the experimental field of Gyeongsangnam-do Agricultural Research and Extension Services in August 2010. The infected fruits showed water-soaked and rot symptoms. White mycelial mats spread over lesions, and then sclerotia were formed on fruit and near soil line. The sclerotia were globoid in shape, 1-3 mm in size and white to brown in color. The optimum temperature for mycelial growth and sclerotia formation on PDA was <TEX>$30^{\circ}C$</TEX> and the hyphal width was 4-8 <TEX>${\mu}m$</TEX>. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. On the basis of mycological characteristics, ITS rDNA sequence analysis, and pathogenicity to host plants, this fungus was identified as Sclerotium rolfsii Saccardo. This is the first report of sclerotium rot on sponge gourd caused by S. rolfsii in Korea.

Ectomycorrhizal iconoclasts: the ITS rDNA diversity and nitrophilic tendencies of fetid Russula

Fetid Russula are frequently dominant ectomycorrhizal fungi, and some appear to be especially nitrophilic. However, little is known about their phylogenetic relationships or how common nitrophilic traits are in this group. This study addresses this gap and presents a phylogenetic analysis of ITS rDNA sequences and a meta-analysis of studies that examine ectomycorrhizal fungi response to nitrogen increase. The phylogenetic analysis indicates that (i) this lineage contains numerous unidentified taxa; (ii) the taxa have distinct geographic distributions; and (iii) the misuse of names such as R amoenolens, R. foetens or R. pectinatoides is common. Twenty-three well supported phylotypes were identified and include clades specific to western North America, eastern North America, Europe and Asia, while morphologically similar collections from tropical-equatorial regions are distinct. The metaanalysis shows that nitrophilic tendencies appear throughout fetid Russulas suggesting that this character is not an isolated trait within this subgenus but instead is a more general feature of the group overall. Mapping these tendencies across a broader portion of the Russulaceae shows that this trait is more regularly found in the basal Russula lineages and Lactarius spp., suggesting that this ability evolved early in these fungi.

Morphological and ITS rDNA-based phylogenetic identification of two new species in Tinctoporellus

Two new poroid wood-inhabiting basidiomycetes, Tinctoporellus bubalinus and T. hinnuleus, are described and illustrated based on morphological characteristics and phylogenetic analysis of rDNA ITS sequences. Tinctoporellus bubalinus is characterized by annual, resupinate basidiocarps, white to buff coloured pore surface and thin- to slightly thick-walled, richly branched and contorted dendrohyphidia. Tinctoporellus hinnuleus is characterized by annual, resupinate basidiocarps, cinnamon, clay-buff to fawn pore surface, thin-walled, richly branched and contorted brown dendrohyphidia. Discriminating characters between the new species and the closely related species are discussed. A key to accepted species in Tinctoporellus is provided.