Elongation Factor 1-alpha Sequences Research Articles

First Report of Root Rot on Rhizome of Polygonatum kingianum Caused by Aspergillus awamori

Polygonatum kingianum Coll. et Hemsl (Huangjing), which belongs to the family Asparagaceae, is a perennial traditional Chinese herb with homologous medicinal and edible value (Liu et al., 2021). Huangjing is known to promote blood circulation; it has anti-inflammatory properties, increases immunity, and provides hypoglycemic treatments (Ma et al., 2019). Root rot-infected P. kingianum exhibited withering yellow leaves and stems, rhizome rot, slowed growth, and plant death. In recent years, with an average incidence of up to 45%, the spread of HJ root rot (rhizome and stem bases) has resulted in a significant reduction in the quality and up to 63% reduction in the yields of Sichuan Junlian (104.5°E, 28.2°N) and Guizhou Zhunyi (107.0°E, 27.7°N). After collecting the diseased samples, we used the tissue isolation method to isolate the pathogenic fungi (Wu et al., 2020). Four fungal isolates associated with root rot were obtained: HJ-G2 (two strains), HJ-G3 (one strain), HJ-G4 (one strain), and HJ-G6 (two strains), of which HJ-G2 and HJ-G6 were the dominant species. To determine pathogenicity of each strain, tests were conducted by wounding rhizomes wth an inoculation needle and the pathogen strain was inoculated onto the wound and symptoms observed. The results reveal that HJ-G6 exhibited the strongest pathogenicity against P. kingianum (Figure 1). The HJ-G6 colonies were black, grew rapidly, and produced a large number of spores (Figure 1). A spherical apical sac (conidial head) is formed at the top with two palisades of cells, metulae and phialides, which are shaped radially and produce a large number of spores with 2-5 um in diameter (Figure 2). Morphological observations revealed that the isolate was consistent with Aspergillus awamori (Naher et al., 2021). To further confirm the fungal species, the ribosomal internal transcribed spacer (ITS), β-tubulin (TUB), and elongation factor 1-alpha (EF-1a) gene regions were amplified with ITS1/ITS4, Bt2a/ Bt2b, and EF1/EF2. Primer and PCR amplification were performed as previously described (Paul et al., 2017). The sequences were compared with those obtained from GenBank. The ITS sequences (GenBank accession number OR682143) of the isolates (HJ-G6) were 100% identical to those of the strain PANCOM10 (GenBank accession number MT007535.1) of Aspergillus awamori. The EF-1a sequences (GenBank accession OR752352) of the isolates (HJ-G6) were 98% identical with strain ITEM 4777 (GenBank accession FN665402.1) of Aspergillus awamori. The TUB sequences (GenBank accession number OR752351) of the isolate (HJ-G6) were 100% identical with strain AF158 (GenBank accession MH781275.1) of Aspergillus awamori. Three maximum likelihood trees were constructed using MEGA v5.0 (Kumar et al., 2018) based on the sequences (ITS, TUB, and EF-1a) of the HJ-G6 strain and that of Aspergillus spp. previously deposited in GenBank (Paul et al., 2017). Phylogenetic analysis showed that HJ-G6 belonged to the Aspergillus awamori clade (Figure 3). Combined with morphological analysis and DNA sequencing, HJ-G6 was identified as Aspergillus awamori. To verify pathogenicity, P. kingianum roots were inoculated with the colonized agar discs of the isolates. P. kingianum plants inoculated with uncolonized agar discs were used as controls. After inoculation, P. kingianum roots were moved to the inoculation chamber under high humidity at 28 °C for 1 d and then transferred to a greenhouse. Previous studies have reported that Fusarium sp. are root rot pathogens in the rhizomes of medicinal plants (Pang et al., 2022; Song et al., 2023). In this study, HJ-G2, HJ-G3, and HJ-G4 were used as the positive controls. Typical symptoms of root rot appeared 3 days after inoculation and were similar to those observed in the field, whereas the control plants remained symptomless. According to the results of the inoculation experiment, the pathogenicity of Aspergillus awamori to P. kingianum root rot was significantly stronger than that of Fusarium (Figure 1). The pathogen was isolated from the rotting root of P. kingianum and the ITS region was sequenced again. Alignment analysis of the ITS sequences revealed that the causal agents were consistent with those of the original isolates. These studies fulfill Koch’s postulates. As far as we know, this is the first report of Aspergillus awamori causing root rot in P. kingianum.

Morphological and phylogenetic analyses reveal four species of myxomycetes new to China.

Four species of myxomycetes (Arcyria pseudodenudata, Diderma europaeum, Lycogala irregulare, and Trichia armillata) new to China were observed via light microscope and scanning electron microscope, and detailed descriptions and illustrations are provided, along with comparisons with related species. Among them, A. pseudodenudata was discovered for the first time outside of the type locality, D. europaeum was discovered for the first time outside of Europe, and L. irregulare and T. armillata were reported again after being named. Phylogenetic analyses based on nuclear 18S rDNA and elongation factor-1 alpha sequences or nuclear 18S rDNA and cytochrome oxidase subunit I sequences was performed to provide a molecular basis for morphological identification. These specimens were deposited in the Herbarium of Fungi of Nanjing Normal University.

Identification and Fungicide Sensitivity of Fusarium spp. Associated with Root Rot of Scutellaria baicalensis in Shanxi Province, China.

Fusarium root rot is usually classified as an extremely destructive soilborne disease. From 2020 to 2021, Fusarium root rot was observed in production areas and seriously affected the yield and quality of Scutellaria baicalensis in Shanxi Province, China. Based on morphological characteristics and combined analysis of the internal transcribed spacer region of ribosomal DNA and translation elongation factor 1-alpha sequences, 68 Fusarium isolates obtained in this work were identified as F. oxysporum (52.94%), F. acuminatum (20.59%), F. solani (16.17%), F. proliferatum (5.88%), F. incarnatum (2.94%), and F. brachygibbosum (1.47%). In the pathogenicity tests, all Fusarium isolates could infect S. baicalensis roots, presenting different pathogenic ability. Among these isolates, F. oxysporum was found to have the highest virulence on S. baicalensis roots, followed by F. acuminatum, F. solani, F.proliferatum, F. brachygibbosum, and F.incarnatum. According to fungicide sensitivity tests, Fusarium isolates were more sensitive to fludioxonil and difenoconazole, followed by carbendazim, thiophanate-methyl, and hymexazol. In brief, this is the first report of Fusarium species (F. oxysporum, F. acuminatum, F. solani, F. proliferatum, F. incarnatum, and F. brachygibbosum) as causal agents of root rot of S.baicalensis in Shanxi Province, China. The fungicide sensitivity results will be helpful for formulating management strategies of S. baicalensis root rot.

First Report of Nigrospora Species Causing Leaf Spot on Olive (Olea europaea L.)

Leaf spot symptoms were spotted in two olive orchards in Istria and in Kvarner Gulf, Croatia. Fungal species from three representative isolates (P13 LECIII, R18 BI, JA20 NP) have been morphologically characterized based on the colony and conidial characteristics. Several techniques were performed for inducing the sporulation of the JA20 NP isolate. Only PDA + banana medium was successful. PCR was conducted for ITS, TUB, and EF1α gene regions. Phylogenetic analyses were performed using internal transcribed spacer, beta-tubulin, and translation elongation factor 1-alpha sequence data. Three types of tests were conducted: a pathogenicity test on detached leaves, on detached and scratched leaves, and on olive seedlings. Ultimately, from the morphological characterizations, DNA sequence analysis of ITS, TUB, and EF1α gene regions, and phylogenetic analysis, these species were identified as Nigrospora gorlenkoana Novobr., Nigrospora osmanthi Mei Wang & L. Cai, and Nigrospora philosophiae-doctoris M. Raza, Qian Chen & L. Cai. This is the first report of Nigrospora species causing leaf spot on olive trees and the first report of Nigrospora philosophiae-doctoris as a plant pathogen. Fungal leaf diseases in conditions that are favorable for infection and disease development can lead to a decrease in the yield and olive oil quality. Therefore, it is necessary to conduct further research and the monitoring of fungal leaf diseases.

Genetic divergence of farmed blue mussels (Mytilus sp.) in Australian waters

As valuable ecosystem engineers, blue mussels, are one of the most ecologically and economically important bivalve species in Australia. The need for understanding levels of genetic diversity in both wild and farmed populations of Australian mussels is ever-growing due to the increased risk of disease and decreased fitness associated with the constantly changing global environment. This study, therefore, investigated the phylogenetic relationship, and the genetic variation of farmed blue mussels (Mytilus sp., previously noted as M. galloprovincialis, M. planulatus or a hybrid of both) at four sites along the south coast of Australia, Victoria, South Australia, Western Australia, and Tasmania. Analyses of two genes, Cytochrome c oxidase subunit 1 gene (mtCOI) and the nuclear gene Elongation factor 1 alpha (EF1a) was completed for all mussels sampled. The phylogenetic reconstruction of mtDNA sequences was not able to cluster the samples into species. The analyses of the nuclear marker (EF1a) returned similar results, except that we could conclude the Australian samples not being M. chilensis. Furthermore, the analyses of EF1a revealed the presence of hybridisation, a pattern commonly observed in mussels. The genetic diversity analysis of mtCOI demonstrated that samples from Victoria and Western Australia had the highest genetic diversity, while South Australia had the lowest. When analysing the nuclear EF1a sequences, mussels from Western Australian had the lowest genetic diversity, and the Victorian samples had the highest.The study extended our knowledge on the population genetic variation of farmed blue mussels (Mytilus sp.) along the south coast of Australia. The identified genetic differences between the four southern states of Australia could be interesting for the aquaculture industry when aiming for more adaptable and profitable stocks.

Ophiocordyceps ovatospora sp. nov. (Ophiocordycipitaceae, Hypocreales), pathogenic on termites from China

A new species, Ophiocordyceps ovatospora, occurring on termites was collected from Yunnan Province, China, and is described based on morphological and molecular phylogenetic data. In a phylogenetic analysis of a combined dataset of multigene loci comprising the internal transcribed spacer regions (ITS), the small and large subunit of the ribosomal DNA (nrSSU and nrLSU), partial sequences of elongation factor 1-alpha (tef-1a) and the largest and second largest subunit of the RNA polymerase (rpb1, rpb2), O. ovatospora was clustered together with O. khokpasiensis to form a clade separate from other species of Ophiocordyceps. Morphologically, O. ovatospora is characterized by producing pseudo-immersed perithecia, filiform asci and ascospores, monophialidic phialides, and oval conidia. Ophiocordyceps ovatospora has smaller perithecia than those of O. khokpasiensis but its asci are larger.

First Report of Diaporthe cercidis Causing Leaf Blotch of Acer pictum subsp. mono in China.

Acer pictum subsp. mono (Maxim.) H. Ohashi is a common deciduous tree species that is widely distributed in Northeast and Northern China, including all provinces of the Yangtze River Basin (Liu et al. 2014). A foliar disease, with an incidence of ~90% (19/21 trees), occurred on A. pictum subsp. mono in a community park, Nanjing, Jiangsu, China in July 2019. On average, ~80% of the leaves per individual tree were infected by this disease. The symptoms initially appeared as brown, necrotic lesions at leaf tips, and half the leaf would become dark brown with time, and finally almost all of leaves were infected. Small pieces of leaf tissue (3 to 5 mm2) cut from the lesion margins were surface-sterilized in 75% ethanol for 30 s and 1% NaClO for 90 s, rinsed with sterile H2O three times, and placed on potato dextrose agar (PDA) at 25°C in the dark. The same fungus was isolated from 92% of the samples. The pure cultures were obtained by single-spore isolation. Three representative isolates (WJF1, WJF3 and WJF4) were obtained, and WJF1 was deposited in China's Forestry Culture Collection Center (CFCC 54806), and WJF3 and WJF4 were deposited at the Nanjing Forestry University (NFU 083 and NFU 084). The culture on PDA was white, with white vigorous aerial mycelia at the edge. Black pycnidia developed on the alfalfa stems at 25°C under a 14/10 h light/dark cycle for 20 days. Conidiophores were hyaline, branched, septate, straight, 16.4-34.7 × 1.5-3.0 μm (n = 30). Conidiogenous cells were 9.0-24.6 × 1.3-2.3 μm (n = 30). Alpha conidia were 7.0 ± 0.6 × 2.2 ± 0.2 μm (n = 30), fusiform, hyaline, smooth and multi-guttulate. Beta conidia were 25.5 ± 4.3 × 1.3 ± 0.1 μm (n = 30), hyaline, smooth and hamate. Morphological characters of all three isolates matched those of Diaporthe spp. (Gomes et al. 2013). DNA of three isolates were extracted and the internal transcribed spacer region (ITS), partial sequences of elongation factor 1-alpha (EF1-α), calmodulin (CAL), beta-tubulin (β-tub) and histone H3 (HIS) genes were amplified with primers ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R and CAL228F/CAL737R (Carbone et al. 1999), βt2a/βt2b and CYLH3F/H3-1b (Glass and Donaldson, 1995, Crous et al. 2004), respectively. The sequences of WJF1, WJF3 and WJF4 were deposited in GenBank (WJF1: Accession Nos. MW301339 for ITS, MW363932 to MW363935 for EF1-α, β-tub, HIS, and CAL; WJF3: MW453062 and MW561566 to MW561569; WJF4: MW453063 and MW561570 to MW561573). BLAST results showed that the ITS, EF1-α, β-tub, HIS, and CAL sequences of WJF1 were similar with sequences of Phomopsis liquidambari C.Q. Chang, Z.D. Jiang & P.K. Chi JQ676191 (identity = 540/540; 100%), D. huangshanensis H. Zhou & C.L. Hou MN224671 (identity = 291/292; 99%), D. pescicola Dissan., J.Y. Yan, Xing H. Li & K.D. Hyde MK691230 (identity = 438/438; 100%), D. spinosa Y.S. Guo & G.P. Wang MK726170 (identity = 437/438; 99%), D. cercidis C.M. Tian & Qin Yang MK691114 (identity = 452/452; 100%), respectively. BLAST results of WJF3 and WJF4 are list in Table 1. A maximum likelihood and Bayesian posterior probability analyses using IQtree v. 1.6.8 and Mr. Bayes v. 3.2.6 with the concatenated sequences placed WJF1, WJF3 and WJF4 in the clade of D. cercidis. Based on the five-locus phylogeny and morphology, WJF1, WJF3 and WJF4 were identified as D. cercidis. The pathogenicity of three isolates were tested on potted 3-yr-old seedlings of A. pictum subsp. mono, grown in a greenhouse. Healthy leaves were wounded with a sterile needle and then inoculated with 10 μL of conidial suspensions (106 conidia/mL). Control leaves were treated with sterilized H2O. A total of twelve seedlings were used for the tests, 3 seedlings per treatment, and five leaves were inoculated per seedling. Each plant was covered with a plastic bag after inoculation and sterilized H2O was sprayed into the bag twice/day to maintain humidity and kept in a greenhouse at the day/night temperatures at 25 ± 2°C/16 ± 2°C. In 5 days, all the inoculated leaves had lesions similar to those observed in the field. D. cercidis was reisolated from the lesions of the inoculated leaves and was confirmed based on morphological characteristics and ITS sequence analysis. No symptoms were observed on the control leaves, and no fungus was isolated from them. D. cercidis was previously reported on twigs of Cercis chinensis (Yang et al. 2018) and causing pear shoot canker (Guo et al. 2020). This is the first report of D. cercidis causing leaf blotch on A. pictum subsp. mono. Identification of the pathogen is imperative for diagnosing and controlling this potentially high risk disease on A. pictum subsp. mono and also for the future studies.

Phylogenetic assessment and taxonomic revision of Halobyssothecium and Lentithecium (Lentitheciaceae, Pleosporales)

Our studies on lignicolous aquatic fungi in Thailand, Sweden, and the UK resulted in the collection of three new Halobyssothecium species (H. bambusicola, H. phragmitis, H. versicolor) assigned to Lentitheciaceae (Pleosporales, Dothideomycetes). Multi-loci phylogenetic analyses of the combined large subunit, small subunit, internal transcribed spacers of ribosomal DNA, and the translation elongation factor 1-alpha sequence data enabled a revision of the taxa assigned to Lentithecium and the transfer of L. cangshanense, L. carbonneanum, L. kunmingense, L. unicellulare, and L. voraginesporum to Halobyssothecium. Collection of an asexual morph of L. lineare and phylogenetic analysis confirmed its taxonomic placement in Keissleriella. Detailed descriptions and illustrations of H. bambusicola, H. phragmitis, and H. versicolor are provided.

First Report of Diaporthe fusicola Causing Leaf Blotch of Osmanthus fragrans in China.

Osmanthus fragrans Lour. is widely distributed in China, Japan, Thailand and India (Zang et al., 2003) and one of the top 10 most well-known flowering plants in China. Since February, 2017, a foliar disease, with a disease incidence of ~60%, occurred on O. fragrans in a community park in Luzhai, Guangxi, China. Symptoms began as round or irregular small yellow spots and became pale brown to gray-brown with time. Small leaf tissues (3 to 4 mm2) cut from lesion margins were surface-sterilized in 75% ethanol for 30 s and 1% NaClO for 90 s before they were rinsed in ddH2O and dried on sterilized filter paper. After drying, the sterilized tissues were plated on potato dextrose agar (PDA) and incubated at 25°C in the dark for 5 days. Five single-spore isolates were obtained and a representative isolate (GH3) was selected and deposited in the China's Forestry Culture Collection Center. The colony on PDA was white with concentric zonation and white aerial mycelia, but the reverse was yellow. Black pycnidia developed on alfalfa extract + Czapek at 25°C with a 14/10 h light/dark cycle after 17 days. Conidiophores were hyaline, branched, septate, straight to sinuous, 12.4-24 × 1.9-2.5 μm (n = 20). The conidia were fusoid, hyaline, smooth, mostly 2-guttules and measured 7.2 ± 0.7 × 2.3 ± 0.2 μm (n = 50). The morphological characters of pycnidia, conidiophores and conidia of all five isolates matched those of Diaporthe spp. (Gomes et al. 2013). DNA of isolates GH3, GH7 and GH8 was extracted and the internal transcribed spacer region (ITS), partial sequences of elongation factor 1-alpha (EF1-α), calmodulin (CAL), beta-tubulin (β-tub) and histone H3 (HIS) genes were amplified with primers ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R and CAL228F/CAL737R (Carbone et al. 1999), βt2a/βt2b and CYLH3F/H3-1b (Glass and Donaldson 1995, Crous et al. 2004), respectively. The sequences of GH3, GH7 and GH8 were deposited in GenBank (GH3: Accession nos. MT499213 for ITS, MT506473 to MT506476 for EF1-α, β-tub, HIS, and CAL; GH7: MT856374 and MT860397 to MT860400; GH8: MT856375 and MT860401 to MT860404). BLAST results showed that the ITS, EF1-α, β-tub, HIS, and CAL sequences of GH3 were highly similar with sequences of Phomopsis sp. [LC168784 (ITS), Identities = 506/506(100%)], Diaporthe fusicola [MK654863 (EF1-α), Identities = 274/275(99%)], D. amygdali [MK570513 (β-tub), Identities = 461/461(100%)], D. fusicola [MK726253 (HIS), Identities = 403/403(100%)] and D. amygdali [KC343263 (CAL), Identities = 428/428(100%)], respectively. A maximum likelihood and Bayesian posterior probability analyses using IQtree v. 1.6.8 and Mr. Bayes v. 3.2.6 with the concatenated sequences placed isolates GH3, GH7 and GH8 in the D. fusicola cluster and separated them from D. eres and D. osmanthi, which were previously reported from Osmanthus spp. (Gomes et al., 2013; Long et al., 2019). Based on the multi-gene phylogeny and morphology, all three isolates were identified as D. fusicola. The pathogenicity of GH3 was tested on 1-yr-old seedlings of O. fragrans. Healthy leaves were wounded with a sterile needle and then inoculated with either 5-mm mycelial plugs cut from the edge of a 5-day-old culture of GH3 or 10 μL of conidial suspensions (106 conidia/mL). Control leaves were treated with PDA plugs or ddH2O. Three plants were used for each treatment. The plants were covered with a plastic bag after inoculation and sterilized H2O was sprayed into the bags twice/day to maintain humidity and kept in a greenhouse at the day/night temperatures at 25 ± 2°C/16 ± 2°C. Lesions appeared 3 days later. No lesions were observed on control leaves. The same fungus was re-isolated from lesions. This is the first report of D. fusicola causing leaf blotch on O. fragrans. These results form the basis for developing effective strategies for monitoring and managing this potential high-risk disease.

New Endophytic Fusarium spp. from Fraxinus excelsior Leaves in Slovakia.

Fraxinus excelsior is currently suffering from ash dieback disease caused by the fungus Hymenoscyphus fraxineus. Co-occurrence of large numbers of other fungi with endophytic as well as pathogenic lifestyles has been observed on F. excelsior. In this study, new endophytic Fusarium spp. isolate Fraxinus excelsior Fusarium from Slovak territory, associated with F.excelsior, was characterised. Using morphology-based approaches, the fungus was assigned to the genus Fusarium. The isolate formed thick-walled hyaline, fusiform, slightly curved macroconidia, produced from monophialides in sporodochia, pointed at the tip, mostly 3 - 5 septate, occasionally 6 septate, and 40 - 68 × 3.8 - 5.0 µm in size with basal pedicellate cells. For more accurate taxonomic affiliation, molecular sequence analyses of the internal transcribed spacer, translation elongation factor 1-alpha sequences, and partial β-tubulin gene were performed. While the internal transcribed spacer sequence of the isolate showed the highest similarity to the Fraxinus oxysporum species, translation elongation factor 1-alpha and partial β-tubulin sequences were distantly related to the Fraxinus avenaceum species. Based on these data, the Fraxinus excelsior Fusarium isolate could probably be considered a new species of the Fusarium genus.

New Molecular Tool for a Quick and Easy Detection of Apple Scab in the Field

Venturia inaequalis, an agent of apple scab, is the most important pathogen of Malus x domestica. Control measures against this pathogen rely on intensive phytosanitary programs based on predictive models to identify the meteorological conditions conducive to the primary infection. The detection of the pathogen in field, both in naturally infected symptomatic and asymptomatic leaves, is desirable. Loop-mediated isothermal amplification (LAMP) assays are profitable molecular diagnostic tools for the direct detection of pathogens in field. A LAMP assay for V. inaequalis has been designed on the elongation factor 1-alpha sequence. The validation of the LAMP assay was carried out following the international EPPO standard PM 7/98 in terms of specificity, sensitivity, repeatability and reproducibility. Specificity testing was performed using target and non-target species, such as phylogenetically related Venturia species and other pathogens commonly found in apple, resulting in positive amplification only for the target with a time to positive ranging from 20 to 30 min. Sensitivity testing was performed with serial dilutions of DNA of the target and by artificial inoculation of young apple leaves. The reliability of the LAMP assay as an early-detection tool and its user-friendly application make it suitable for the diagnosis of apple scab in the field.

Identification of the pathogens causing konjac dry rot in China

Dry rot causes significant losses of konjac during storage in Hubei Province, China. In this study, the pathogens were obtained from konjac and then tested its pathogenicity through tissue isolation. The pathogens were identified using morphological characteristics and translation elongation factor-1 alpha sequences analysis. Four Fusarium species, namely F. meridionale, F. incarnatum, F. sambucinum, and F. solani, were obtained.

The cup fungus Pestalopezia brunneopruinosa is Pestalotiopsis gibbosa and belongs to Sordariomycetes.

Pestalopezia brunneopruinosa, the type species of Pestalopezia in Leotiomycetes, produces typical cup-shaped ascomata. Because its asexual morph has conidia comprised of five cells including apical and basal appendages and three pigmented median cells, it was first described as Pestalotia gibbosa, which belongs to Sordariomycetes. This contradiction has not been resolved due to the difficulty in isolating this fungus in culture. In this study, we isolated separate strains from the sexual morph and the asexual morph for molecular analysis. Phylogenetic trees of Sporocadaceae based on internal transcribed spacer, partial β-tubulin, and partial translation elongation factor 1-alpha sequence datasets revealed that both strains fall into the same taxon, in a clade in Pestalotiopsis sensu stricto alongside P. gaultheriae and P. spathulata. We provide the first evidence that fungi producing cup-shaped ascomata in Pestalotiopsis belong to Sordariomycetes, and we have proposed the transfer of Pestalopezia brunneopruinosa to Pestalotiopsis gibbosa.

Morosphaeria muthupetensis sp. nov. (Morosphaeriaceae) from India: morphological characterization and multigene phylogenetic inference

Abstract A novel species of Morosphaeria, Morosphaeria muthupetensis, saprobic on intertidal mangrove wood of Rhizophora mucronata from Muthupet mangroves on Kaveri River Delta, Tamil Nadu along the East coast of India, is described in this paper. Morosphaeria muthupetensis is characterized by large sub-globose to lenticular ascomata; long cylindrical to clavate, short pedicellate asci; small, fusiform to ellipsoidal ascospores with acute ends when compared to other Morosphaeria species. A multigene phylogeny obtained from a combined nuclear large subunit, small subunit, internal transcribed spacers rDNA and translation elongation factor 1-alpha sequence analysis revealed that M. muthupetensis is phylogenetically distinct and nested in between Morosphaeria velatispora and Morosphaeria ramunculicola and all these species constitute a strongly supported monophyletic clade sister to the genus Helicascus. Detailed descriptions and illustrations of M. muthupetensis are presented and compared with other Morosphaeria species.

Novel and interesting Ophiocordyceps spp. (Ophiocordycipitaceae, Hypocreales) with superficial perithecia from Thailand.

Ophiocordyceps is a heterogeneous, species-rich genus in the order Hypocreales (Sordariomycetes, Ascomycota) that includes invertebrate-pathogenic taxa. In this study, seven new species in Ophiocordyceps producing superficial perithecia infecting various insect hosts (Lepidoptera, Hemiptera) are described from Thailand – Ophiocordyceps brunneinigra, O. brunneiperitheciata, O. geometridicola, O. multiperitheciata, O. pauciovoperitheciata, O. pseudoacicularis and O. spataforae. Phylogenetic analyses based on multigene loci comprising the large subunit of the ribosomal DNA (LSU), partial sequences of elongation factor 1-alpha (TEF) and the largest and second largest subunit of the RNA polymerase (RPB1, PRB2) strongly support these new species of Ophiocordyceps in the Ophiocordycipitaceae. They differ from species previously described species Ophiocordyceps acicularis, O. atewensis, O. cochlidiicola, and O. crinalis, in the shape and sizes of distinguishing characters such as perithecia, ascospores and conidia. We also report a new record of O. macroacicularis in Thailand.

Speciation in progress? A phylogeographic study among populations of Hemitrichia serpula (Myxomycetes).

Myxomycetes (plasmodial slime molds, Amoebozoa) are often perceived as widely distributed, confounding to the “everything is everywhere” hypothesis. To test if gene flow within these spore-dispersed protists is restricted by geographical barriers, we chose the widespread but morphologically unmistakable species Hemitrichia serpula for a phylogeographic study. Partial sequences from nuclear ribosomal RNA genes (SSU) revealed 40 ribotypes among 135 specimens, belonging to three major clades. Each clade is dominated by specimens from a certain region and by one of two morphological varieties which can be differentiated by SEM micrographs. Partial sequences of the protein elongation factor 1 alpha (EF1A) showed each clade to possess a unique combination of SSU and EF1A genotypes. This pattern is best explained assuming the existence of several putative biospecies dominating in a particular geographical region. However, occasional mismatches between molecular data and morphological characters, but as well heterogeneous SSU and heterozygous EF1A sequences, point to ongoing speciation. Environmental niche models suggest that the putative biospecies are rather restricted by geographical barriers than by macroecological conditions. Like other protists, myxomycetes seem to follow the moderate endemicity hypothesis and are in active speciation, which is most likely shaped by limited gene flow and reproductive isolation.

A new species of Cantharellus (Cantharellales, Basidiomycota, Fungi) from subalpine forest in Yunnan, China

Cantharellus versicolor is described and illustrated as a new species based on morphological and molecular characters. The most significant features to distinguish the new species from other known Cantharellus are its extremely fleshy, turning gray after injury and with black floccose-fibrillose scales composed of thick-walled and irregular, erect hyphae on the pileus. It is described from the subalpine belt of Shangri-La, northwestern Yunnan, China. Phylogenetic analysis of the transcription elongation factor 1-alpha sequence data further support its systematic position in the subgenus Cantharellus and its description as a new species.

First Report of Fusarium subglutinans Causing Leaf Spot Disease on Cymbidium Orchids in Korea

In 2006~2010, leaf spot symptoms, that is, small, yellow spots that turned into dark brown-to-black lesions surrounded by a yellow halo, were observed on Cymbidium spp. in Gongju, Taean, and Gapyeong in Korea. A Fusarium species was continuously isolated from symptomatic leaves; in pathogenicity testing, isolates caused leaf spot symptoms consisting of sunken, dark brown lesions similar to the original ones. The causal pathogen was identified as Fusarium subglutinans based on morphological and translation elongation factor 1-alpha sequence analyses. This is the first report of F. subglutinans as the cause ofleaf spot disease in Cymbidium spp. in Korea.

Phylogeny and taxonomy of species in the Grosmannia serpens complex

Grosmannia serpens was first described from pine in Italy in 1936 and it has been recorded subsequently from many countries in both the northern and southern hemispheres. The fungus is vectored primarily by root-infesting bark beetles and has been reported to contribute to pine-root diseases in Italy and South Africa. The objective of this study was to consider the identity of a global collection of isolates not previously available and using DNA sequence-based comparisons not previously applied to most of these isolates. Phylogenetic analyses of the ITS2-LSU, actin, beta-tubulin, calmodulin and translation elongation factor-1 alpha sequences revealed that these morphologically similar isolates represent a complex of five cryptic species. Grosmannia serpens sensu stricto thus is redefined and comprises only isolates from Italy including the ex-type isolate. The ex-type isolate of Verticicladiella alacris was shown to be distinct from G. serpens, and a new holomorphic species, G. alacris, is described. The teleomorph state of G. alacris was obtained through mating studies in the laboratory, confirming that this species is heterothallic. Most of the available isolates, including those from South Africa, USA, France, Portugal and some from Spain, represent G. alacris. The remaining three taxa, known only in their anamorph states, are described as the new species Leptographium gibbsii for isolates from the UK, L. yamaokae for isolates from Japan and L. castellanum for isolates from Spain and the Dominican Republic.

First report of Fusarium redolens from Saskatchewan and its comparative pathogenicity

Fusarium redolens [syn: F. oxysporum var. redolens] is pathogenic on a wide range of plant species. Fusarium redolens was frequently isolated from necrotic and discolored root and crown tissues of chickpea, pea, lentil and durum wheat in Saskatchewan. The fungus was identified using distance analysis of the translation elongation factor-1 alpha sequences. Comparative pathogenicity tests showed that F. redolens, like Fusarium graminearum and Cochliobolus sativus, caused lesions and/or discolorations on the root, crown and shoot of durum wheat, pea and chickpea, although it had a different host preference. While F. redolens was most virulent on pea, the other two fungi were more virulent on durum wheat. It appears that F. redolens is common in the Prairie Ecozone of Saskatchewan and it may cause yield reduction in pea, chickpea and durum crops. This warrants a closer look at the biology and ecology of F. redolens in the Canadian Prairies.