LSU rRNA Gene Research Articles

(GTG)5 MSP-PCR fingerprinting as a technique for discrimination of wine associated yeasts?

In microbiology, identification of all isolates by sequencing is still unfeasible in small research laboratories. Therefore, many yeast diversity studies follow a screening procedure consisting of clustering the yeast isolates using MSP-PCR fingerprinting, followed by identification of one or a few selected representatives of each cluster by sequencing. Although this procedure has been widely applied in the literature, it has not been properly validated. We evaluated a standardized protocol using MSP-PCR fingerprinting with the primers (GTG)5 and M13 for the discrimination of wine associated yeasts in South Brazil. Two datasets were used: yeasts isolated from bottled wines and vineyard environments. We compared the discriminatory power of both primers in a subset of 16 strains, choosing the primer (GTG)5 for further evaluation. Afterwards, we applied this technique to 245 strains, and compared the results with the identification obtained by partial sequencing of the LSU rRNA gene, considered as the gold standard. An array matrix was constructed for each dataset and used as input for clustering with two methods (hierarchical dendrograms and QAPGrid layout). For both yeast datasets, unrelated species were clustered in the same group. The sensitivity score of (GTG)5 MSP-PCR fingerprinting was high, but specificity was low. As a conclusion, the yeast diversity inferred in several previous studies may have been underestimated and some isolates were probably misidentified due to the compliance to this screening procedure.

Wickerhamiella siamensis f.a., sp. nov., an endophytic and epiphytic yeast species isolated from sugar cane leaf.

Six strains representing a novel yeast species were isolated from tissue (DMKU-SE106(T), DMKU-SE110, DMKU-SE112 and DMKU-SE132) and the external surface (DMKU-SP335 and DMKU-SP406) of sugar cane leaves collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and sequence analysis of the D1/D2 region of the LSU rRNA gene and the internal transcribed spacer (ITS) region, the six strains were found to represent a single novel species of the genus Wickerhamiella although the formation of ascospores was not observed. The sequences of the D1/D2 region of the LSU rRNA gene and ITS region of the six strains differed from each other by 0-2 and 2-3 nt substitutions, respectively. The novel species was related most closely to Candida infanticola but with 4.5-4.6% nucleotide substitutions in the D1/D2 region of the LSU rRNA gene and 6.6-7.1% nucleotide substitutions in the ITS region. The name Wickerhamiella siamensis f.a., sp. nov. is proposed. The type strain is DMKU-SE106(T) ( =BCC 61185(T) =NBRC 109697(T) =CBS 13331(T)).

Biogeography and Phylogenetic Position of a Warm-stenotherm Centric Diatom, Skeletonema potamos (C.I. Weber) Hasle and its Long-term Dynamics in the River Danube

Skeletonema potamos is a poorly known freshwater species in the ancestrally and predominantly marine genus Skeletonema. With phylogenetic analysis of two nuclear {partial SSU (18S) and partial LSU (28S) rDNA)} and two chloroplast (rbcL and psbC) genes, we verified its placement within the genus Skeletonema and identified the mostly brackish species, Skeletonema subsalsum, as its closest known relative. Comparisons of SSU and LSU rRNA genes from S. potamos populations from Europe and North America revealed no intraspecific variation. Skeletonema potamos can be a dominant element of the phytoplankton community in various ecosystems, including the River Danube. We tracked phytoplankton composition in the River Danube weekly from 1979 to 2 012, and throughout this period, S. potamos exhibited a strong increase in proportion of total phytoplankton abundance and biomass - an increase that was correlated with increasing water temperature over the same time period. Current records indicate a temperate distribution of S. potamos, but ecological data predict possible expansion of its geographic range and increase in seasonal duration within existing habitats in response to the warming of surface waters.

The more we search, the more we find: discovery of a new lineage and a new species complex in the genus Asparagopsis.

In the past few decades, in the marine realm in particular, the use of molecular tools has led to the discovery of hidden taxonomic diversity, revealing complexes of sister species. A good example is the red algal genus Asparagopsis. The two species (A. armata and A. taxiformis) recognized in this genus have been introduced in many places around the world. Within the nominal species A. taxiformis, previous molecular analyses have uncovered several lineages, suggesting the existence of sister species or subspecies. Although the genus has been well studied in some regions (e.g., the Mediterranean Sea and Hawaii), it remains poorly investigated in others (e.g., South Pacific). Our study mainly focused on these latter areas to clarify lineages and better determine lineage status (i.e., native vs. introduced). A total of 188 specimens were collected from 61 sites, 58 of which had never been sampled before. We sequenced the DNA from samples for three markers and obtained 112 sequences for the chloroplastic RuBisCo spacer, 118 sequences for the nuclear LSU rRNA gene, and 174 for the mitochondrial spacer cox2-3. Phylogenetic analyses using all three markers suggested the existence of two cryptic sister species with the discovery of a new clade within A. armata. This clade was found only in Western Australia, Tasmania and New Zealand, and is thus restricted to a subregional biogeographic unit. We also discovered a new, fifth lineage for A. taxiformis restricted to the South Pacific and Western Australia. Except for this newly described lineage, all other lineages showed a global distribution influenced by introduction events. These results illustrate the difficulty in accurately defining cosmopolitan species. Our findings also highlight the need for targeted (i.e., in poorly studied areas) and geographically extensive sampling efforts when studying taxa that have been introduced globally and that are likely to hide species complexes.

High fungal diversity and abundance recovered in the deep-sea sediments of the Pacific Ocean.

Knowledge about the presence and ecological significance of bacteria and archaea in the deep-sea environments has been well recognized, but the eukaryotic microorganisms, such as fungi, have rarely been reported. The present study investigated the composition and abundance of fungal community in the deep-sea sediments of the Pacific Ocean. In this study, a total of 1,947 internal transcribed spacer (ITS) regions of fungal rRNA gene clones were recovered from five sediment samples at the Pacific Ocean (water depths ranging from 5,017 to 6,986 m) using three different PCR primer sets. There were 16, 17, and 15 different operational taxonomic units (OTUs) identified from fungal-universal, Ascomycota-, and Basidiomycota-specific clone libraries, respectively. Majority of the recovered sequences belonged to diverse phylotypes of Ascomycota (25 phylotypes) and Basidiomycota (18 phylotypes). The multiple primer approach totally recovered 27 phylotypes which showed low similarities (≤97 %) with available fungal sequences in the GenBank, suggesting possible new fungal taxa occurring in the deep-sea environments or belonging to taxa not represented in the GenBank. Our results also recovered high fungal LSU rRNA gene copy numbers (3.52 × 10(6) to 5.23 × 10(7)copies/g wet sediment) from the Pacific Ocean sediment samples, suggesting that the fungi might be involved in important ecological functions in the deep-sea environments.

Validation of the detection of Pseudo-nitzschia spp. using specific RNA probes tested in a microarray format: Calibration of signal based on variability of RNA content with environmental conditions

Harmful algal blooms (HAB) occur worldwide and cause health problems and economic damage to fisheries and tourism. Monitoring for toxic algae is therefore essential but is based primarily on light microscopy, which is time consuming and can be limited by insufficient morphological characters such that more time is needed to examine critical features with electron microscopy. Monitoring with molecular tools is done in only a few places world-wide. EU FP7 MIDTAL (Microarray Detection of Toxic Algae) used SSU and LSU rRNA genes as targets on microarrays to identify toxic species. In order to comply with current monitoring requirements to report cell numbers as the relevant threshold measurement to trigger closure of fisheries, it was necessary to calibrate our microarray to convert the hybridisation signal obtained to cell numbers. Calibration curves for two species of Pseudo-nitzschia for use with the MIDTAL microarray are presented to obtain cell numbers following hybridisation. It complements work presented by Barra et al. (2012b. Environ. Sci. Pollut. Res. doi: 10.1007/s11356-012-1330-1v) for two other Pseudo-nitzschia spp., Dittami and Edvardsen (2012a. J. Phycol. 48, 1050) for Pseudochatonella, Blanco et al. (2013. Harmful Algae 24, 80) for Heterosigma, McCoy et al. (2013. FEMS. doi: 10.1111/1574-6941.12277) for Prymnesium spp., Karlodinium veneficum, and cf. Chatonella spp. and Taylor et al. (2014. Harmful Algae, in press) for Alexandrium.

Surface soil fungal and bacterial communities in aspen stands are resilient to eleven years of elevated CO2 and O3

Soil microbial community changes associated with altered carbon cycling dynamics were investigated in aspen (Populus tremuloides) stands exposed to elevated atmospheric CO2 (eCO2) and/or elevated ozone (eO3) for eleven years, at the Rhinelander Free Air CO2 Enrichment (FACE) experiment (Wisconsin, USA). The relative abundance and composition of fungal and bacterial communities in the 0–5 cm soil depth was evaluated using qPCR of fungal or bacterial rRNA genes, cloned 16S (bacterial) and LSU (fungal) rRNA gene sequencing, small subunit rRNA pyrosequencing, and soil shotgun metagenomes. Fungal or bacterial biomass, measured by qPCR, did not differ with treatment. Under eO3, the ratio of Basidiomycota to Ascomycota significantly increased by two-fold (P < 0.04) relative to the ambient control, but no robust significant differences (P > 0.1) in the ß diversity of fungal or bacterial communities were detected under eCO2. The overall composition of fungal and bacterial communities was resilient despite substantial changes in plant productivity, plant genotype, root density, and rates of soil C and N cycling under eCO2, documented in prior studies. Nonetheless, some candidate taxa (up to 20% of the total at the genus level) shifted in relative abundance in soils from eCO2 plots, suggesting that although community composition was generally stable, it was not static. The eCO2-responsive taxa did not shift under eCO2 plus eO3 conditions, suggesting an interactive effect of eO3 on the soil community. Our results show that the fungal and bacterial communities in the 0–5 cm depth mineral soil changed only in small ways after eleven years of eCO2. This study reinforces the view that soil microbial community changes in ecosystems responding to global climate change drivers may be highly-localized in soil strata.

Hagleromyces gen. nov., a yeast genus in the Saccharomycetaceae, and description of Hagleromyces aurorensis sp. nov., isolated from water tanks of bromeliads.

Three strains of a novel yeast species were isolated from water tanks (phytotelmata) of a bromeliad species collected in the state of Tocantins, Brazil. Analysis of sequences for the region spanning the SSU rRNA gene, the internal transcribed spacer, the 5.8S rRNA gene and the D1/D2 domains of the LSU rRNA gene and RNA polymerase II gene showed that these novel yeasts belong to a species that is distinct from all recognized ascomycetous yeast species. Based on the results of gene sequence analyses, a novel species representing a new genus in the Saccharomycetaceae is proposed. The novel species is assigned to the genus Hagleromyces gen. nov. The three isolates of the novel yeast species failed to form sexual spores alone or in mixtures. The name Hagleromyces aurorensis sp. nov. is proposed to accommodate these isolates. The type strain of H. aurorensis sp. nov. is UFMG-CM-Y311(T) ( = CBS 13264(T)).

Fungal community structure in disease suppressive soils assessed by 28S LSU gene sequencing.

Natural biological suppression of soil-borne diseases is a function of the activity and composition of soil microbial communities. Soil microbe and phytopathogen interactions can occur prior to crop sowing and/or in the rhizosphere, subsequently influencing both plant growth and productivity. Research on suppressive microbial communities has concentrated on bacteria although fungi can also influence soil-borne disease. Fungi were analyzed in co-located soils ‘suppressive’ or ‘non-suppressive’ for disease caused by Rhizoctonia solani AG 8 at two sites in South Australia using 454 pyrosequencing targeting the fungal 28S LSU rRNA gene. DNA was extracted from a minimum of 125 g of soil per replicate to reduce the micro-scale community variability, and from soil samples taken at sowing and from the rhizosphere at 7 weeks to cover the peak Rhizoctonia infection period. A total of ∼994,000 reads were classified into 917 genera covering 54% of the RDP Fungal Classifier database, a high diversity for an alkaline, low organic matter soil. Statistical analyses and community ordinations revealed significant differences in fungal community composition between suppressive and non-suppressive soil and between soil type/location. The majority of differences associated with suppressive soils were attributed to less than 40 genera including a number of endophytic species with plant pathogen suppression potentials and mycoparasites such as Xylaria spp. Non-suppressive soils were dominated by Alternaria, Gibberella and Penicillum. Pyrosequencing generated a detailed description of fungal community structure and identified candidate taxa that may influence pathogen-plant interactions in stable disease suppression.

Wickerhamiella kiyanii f.a., sp. nov. and Wickerhamiella fructicola f.a., sp. nov., two yeasts isolated from native plants of Atlantic rainforest in Brazil

Two novel species, Wickerhamiella kiyanii f.a., sp. nov. (type strain FB1-1DASP(T) = CBS 12905(T) = CBMAI 1613(T)) and Wickerhamiella fructicola f.a., sp. nov. (type strain H10Y(T) = CBS 12902(T) = CBMAI 1614(T)) are proposed in the Wickerhamiella clade (Saccharomycetes, Saccharomycetales) to accommodate three strains isolated from flowers and fruits typical of the Brazilian Atlantic rainforest. The novel status of these yeast species was established by sequence divergence observed in the D1/D2 domains of the LSU rRNA gene from the most closely related, described species as well as by phylogenetic analysis. Wickerhamiella kiyanii sp. nov. differs from its nearest phylogenetic neighbours W. pagnoccae CBS 12178(T), Candida jalapaonensis CBS 10935(T) and Candida drosophilae CBS 8459(T) by 2.2-4.2% in the D1/D2 sequences. By contrast, a sequence divergence of 13.2-13.8% was observed between W. fructicola sp. nov. and its closest, described phylogenetic relative Candida kazoui JCM 12558(T) and Candida hasegawae JCM 12559(T). Taxonomic descriptions of the two novel species are given.

Starmerella aceti f.a., sp. nov., an ascomycetous yeast species isolated from fungus garden of the leafcutter ant Acromyrmex balzani

A novel yeast species was recovered from the fungus garden of the leaf-cutting ant Acromyrmex balzani (Hymenoptera: Formicidae). The growth of the novel yeast species is limited by its ability to metabolize only a few carbon and nitrogenous compounds. A remarkable characteristic of this strain is the vigorous growth in 1 % acetic acid. Sequence analysis of the D1/D2 domains of the LSU rRNA gene showed that the novel species belongs to the Starmerella clade and is phenotypically and genetically divergent from currently recognized species in this clade. Described here as Starmerella aceti f.a., sp. nov., it differs by 37 nucleotide substitutions in the D1/D2 region from Starmerella jinningensis CBS 11864(T), the most closely related species. The type strain of Starmerella aceti sp. nov. is TO 125(T) ( = CBMAI 1594(T) = CBS 13086(T)).

Wickerhamomyces arborarius f.a., sp. nov., an ascomycetous yeast species found in arboreal habitats on three different continents

Five strains representing a novel yeast species belonging to the genus Wickerhamomyces were independently isolated from Ecuador, Taiwan and the USA. One strain (CLQCA 10-161(T)) was isolated from the white flower of an unidentified plant species collected in the Maquipucuna cloud forest reserve, near Quito, in Ecuador. A second strain (GY7L12) was isolated from the leaf of a Chinese sumac or nutgall tree (Rhus chinensis 'roxburghiana') collected in the Taoyuan mountain area, Kachsiung, in Taiwan. Three additional strains (A543, A546 and A563) were isolated from two species of wood-boring beetle (Xyleborus glabratus and Xyleborinus saxeseni) collected near Clyo, Georgia, USA. Analysis of the D1/D2 domains of the LSU rRNA gene indicated that the novel species belongs to the genus Wickerhamomyces, and is most closely related to Wickerhamomyces sydowiorum, an insect-associated species predominantly found in South Africa. The North American and Taiwanese strains have identical internal transcribed spacer (ITS) sequences and can be distinguished from the Ecuadorian strain based on a single nucleotide substitution in the ITS1 region. The species name of Wickerhamomyces arborarius f.a., sp. nov. is proposed to accommodate these strains, with CLQCA 10-161(T) ( = CBS 12941(T) = NCYC 3743(T)) designated the type strain.

Yamadazyma ubonensis f.a., sp. nov., a novel xylitol-producing yeast species isolated in Thailand

Three hundred and thirty-seven xylose-utilizing yeast strains were isolated from various natural samples. Among these, 68 strains produced xylitol in the range of 0.1-0.69 g xylitol/g xylose. Thirty-nine xylitol-producing strains were identified to be Candida tropicalis. Ten strains were found belonging to 14 known species in the genus Candida, Cyberlindnera, Meyerozyma, Pichia, Wickerhamomyces, Yamadazyma and Cryptococcus. Two strains were identified to be two Candida species and two strains (DMKU-XE142(T) and DMKU-XE332) were found to be a novel species. Strain DMKU-XE142(T) was isolated from tree bark and DMKU-XE332 was obtained from decaying plant leaf collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics and sequence analysis of the D1/D2 region of the large subunit rRNA gene (LSU) and the internal transcribed spacer (ITS) region, the two strains were determined to represent a novel Yamadazyma species although formation of ascospores was not observed. The sequences of the D1/D2 region of the LSU rRNA gene and the ITS region of the two strains were identical but differed from Yamadazyma phyllophila, the closest species in terms of pairwise sequence similarity of the D1/D2 region, by 1.7 % nucleotide substitutions and 3.5 % nucleotide substitutions in the ITS region. The name Yamadazyma ubonensis f.a., sp. nov. is proposed (type strain is DMKU-XE142(T) = BCC 61020(T) = CBS 12859(T)).

Molecular evidence confirms the parasite Congracilaria babae (Gracilariaceae, Rhodophyta) from Malaysia

A phylogenetic study of two parasites found on the common host species Gracilaria salicornia from Japan and Malaysia based on nuclear, plastid, and mitochondrial molecular markers was conducted. The Japanese and Malaysian parasites were placed in the same cluster in the nuclear phylogenies inferred from the LSU rRNA gene and ITS region, supporting the monophyly of these parasites despite the slight anatomical variations observed. In addition to the molecular analyses, shared morphological and reproductive features including the lack of rhizoids, similar coloration to hosts, and the presence of deep spermatangial conceptacles supported the inclusion of both the Japanese and Malaysian parasites in Congracilaria babae. Phylogenetic analyses based on ITS sequences indicated that C. babae most likely evolved directly from its current host species, G. salicornia. C. babae was shown to be a member of Gracilaria sensu stricto along with its host. C. babae growing on G. salicornia is characterized by having plastid rbcL and mitochondrial cox1 gene sequences identical to those of its host, whereas the nuclear LSU rRNA gene and ITS region sequences are different from those of its host.

Yarrowia divulgata f.a., sp. nov., a yeast species from animal-related and marine sources

Five yeast strains, phenotypically indistinguishable from Yarrowia lipolytica and Yarrowia deformans, were recovered from different animal-related samples. One strain was isolated from a bacon processing plant in Denmark, two strains from chicken liver in the USA, one strain from chicken breast in Hungary and one from minced beef in Hungary. Comparisons of the sequences of their large subunit rRNA gene D1/D2 domain and the internal transcribed spacer (ITS) regions revealed that, despite their phenotypic similarity, they represent a novel yeast species of the Yarrowia clade with Y. deformans being the genotypically closest relative (LSU rRNA gene D1/D2 and ITS region similarity of 97.0 and 93.7 %, respectively). Yarrowia divulgata f.a., sp. nov. is proposed to accommodate these strains with F6-17(T) ( = CBS 11013(T) = CCUG 56725(T)) as the type strain. Some D1/D2 sequences of yeasts from marine habitats were found in the GenBank database that were identical to those of the strains of Y. divulgata f.a., sp. nov. Unfortunately, these strains were not available for our study.

Nakazawaea siamensis f.a., sp. nov., a yeast species isolated from phylloplane

Strain DMKU-RK467(T), representing a novel yeast species, was isolated from the external surface of sugar cane leaves collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and sequence analysis of the D1/D2 region of the LSU rRNA gene and the internal transcribed spacer (ITS) region, strain DMKU-RK467(T) was assigned to a novel species of the genus Nakazawaea. The novel species was related most closely to the type strain of Candida wickerhamii but they differed by 1.9 % nucleotide substitutions in the D1/D2 region of the LSU rRNA gene and by 5.2 % nucleotide substitutions in the ITS region. The name Nakazawaea siamensis f.a., sp. nov. is proposed (type strain DMKU-RK467(T) = BCC 50734(T) = NBRC 108903(T) = CBS 12569(T)).

Dyfrolomycetaceae, a new family in the Dothideomycetes, Ascomycota

A new mangrove fungus collected in Tioman Island, Malaysia, is morphologically similar to marine species of Saccardoella. It also phylogenetically groups with Saccardoella rhizophorae in the Dothideomycetes, based on combined analysis of partial SSU, LSU rRNA and TEF1 gene sequences. The new fungus and S. rhizophorae form a well-supported clade with Acrospermum spp. in the Acrospermaceae. Both species therefore do not belong in Saccardoella, a genus with unitunicate asci. A new genus, Dyfrolomyces, is established to accommodate the new fungus (Dyfrolomyces tiomanensis) while the three marine Saccardoella species (S. mangrovei, S. marinospora, S. rhizophorae) are transferred to the new genus. Dyfrolomyces is characterized by forming a clypeus on substrates, with immersed perithecial ascomata, bitunicate/fissitunicate asci and multi-septate ascospores with/without a sheath. Since D. rhizophorae and D. tiomanensis do not cluster with any known families in the Dothideomycetes, a new family, Dyfrolomycetaceae, is introduced to accommodate the Dyfrolomyces species.

Kazachstania psychrophila sp. nov., a novel psychrophilic yeast isolated from vacuum-packed beef

Five novel ascosporogenous yeast strains (H382, H396, H409, H433(T) and H441) were found through a survey of vacuum-packed beef microbiota. Sequence analysis of ITS domain and LSU rRNA genes showed that the new strains represent a distinct lineage within the genus Kazachstania, closely related to Kazachstania lodderae (97.0 % identity) and Kazachstania ichnusensis (96.1 % identity). The main difference of strains H382, H396, H409, H433(T) and H441 to strains of known Kazachstania species is the maximum growth temperature, which is below 20 °C for the new strains, whereas related species grow at 25 °C. Furthermore, the strains differed from known Kazachstania species in assimilation and fermentation patterns of carbon sources. Based on these characteristics, the five strains are considered to represent a novel species of the genus Kazachstania for which the name Kazachstania psychrophila sp. nov. is proposed. The type strain is H433(T) (DSM 26230(T)=CBS 12689(T)). The Mycobank number of the type strain is MB 803980.

Rate of Pneumocystis pneumonia in Iranian HIV+ Patients with Pulmonary Infiltrates

Background: Pneumocystis jirovecii (formerly known as P. carinii) is one of the most opportunistic agents, which frequently leads to hospitalization and death in immunocompromised patients, especially in HIV-infected individuals. Objectives: The current study is the first report on the rate of the infection in Iranian HIV-infected individuals. Patients and Methods: We used two nested PCR and PCR-RFLP assays to amplify mt LSU rRNA and DHPS genes in 126 samples obtained from the respiratory systems of Iranian HIV-patients with CD4 count < 200 cells/μL, who were referred to two Research Centers from August 2010 to March 2011. Results: In the group of studied patients, 112 were male (88.9%). The mean age of the patients was 35.12 ± 9.75 years. Median CD4 T cell count of the patients was 93 cells/μL. Thirty nine patients (31%) were hospitalized, and 24 patients (18.9%) tested positive for Mycobacterium tuberculosis. In spite of receiving Pneumocystis pneumonia prophylaxis and highly active anti-retroviral therapy, P. jirovecii was detected in 15 samples (11.9%). Just one patient (0.8%) was co-infected with Mycobaterium tuberculosis. The mortality rate due to Pneumocystis pneumonia (PCP) was 26.6%. None of the isolates showed any mutation in codons 55 and 57, which are associated with resistance to sulfa/sulfon drugs. The sequencing results showed that the genetic patterns of Iranian isolates were similar to Indian and Spanish isolates (99% identity). Conclusions: The study indicated that the rate of P. jirovecii infection is similar to those reported from Asian, Indian and African countries. The similarity of the genetic pattern between Iranian and Indian isolates is probably due to their close geographical proximity.

Two new Ramicandelaber species from Taiwan

Two new species of Ramicandelaber isolated from soil in Taiwan are described. Ramicandelaber fabisporus sp. nov. is characterized by having bean-shaped spores that are the smallest in the genus and also by the absence of lateral branches on the sporangiophores. Ramicandelaber taiwanensis sp. nov. is characterized by small, fusiform spores and it produces lateral branches that arise from the sporangiophores. Morphological characters and molecular phylogenetic analysis (D1/D2 domains of the LSU rRNA genes and ITS region) justify these new species of Ramicandelaber. A key is provided to the recognized species of Ramicandelaber.