ITS Analysis Research Articles

Botryosphaeria dothidea causing leaf blight of Photinia serrulata in China

Photinia serrulata is a widely cultivated ornamental plant in China and many other nations. P. serrulata leaf blight was observed in April 2022 in Xinxiang City, Henan Province, China. On the leaves, typical leaf blight symptoms included brown necrotic lesions with alternating rings. Botryosphaeria dothidea was isolated from symptomatic leaves of P. serrulata and identified through morphological and phylogenetic analysis of ITS, TEF1-α, and TUB2. According to the biological characteristics of B. dothidea, the optimal growth temperature and pH are 30 °C and 6, respectively. In addition, light conditions can promote the growth of mycelium. Under controlled environmental conditions, P. serrulata detached leaves and seedlings were subjected to inoculation experiments. Inoculation with a B. dothidea isolate produced typical symptoms of brown blight, and the same pathogen was re-isolated and identified. According to our knowledge, this is the first report of leaf blight disease caused by B. dothidea on P. serrulata in China.

Physiological and Biochemical characters of <i>Nigrospora gorlenkoana</i> Novobr. Occurring on Cereals

The ubiquitous distribution of Nigrospora fungi in the mycobiota of cereal grain in Russia has been established. Fourteen Nigrospora strains using phylogenetic analysis of ITS, tub, and TEF loci were identified as N. gorlenkoana, first identified in Russia. Morphological characteristics of N. gorlenkoana strains were analyzed on various nutrient media. A wide temperature range of 15–25°C was optimal for growth of N. gorlenkoana strains. This probably contributes to the wide distribution of N. gorlenkoana in nature. The pathogenicity of N. gorlenkoana strains to cereal crops was shown. The fungal strains reduced length of wheat and barley seedlings on average 48% and 39% compared to the control, respectively.

Morphology and multi-gene phylogeny reveal a novel Torula (Pleosporales, Torulaceae) species from the plateau lakes in Yunnan, China.

During an investigation into lignicolous freshwater fungi from the plateau lakes in Yunnan Province, China, two fresh collections of Torula taxa were collected and examined morpholgically. Torulaluguhuensis is characterised by: conidiophores which are semi-macronematous mononematous, erect, septate, smooth, slightly flexuous and pale brown; conidiogenous cells which are holoblastic, mono- to polyblastic, integrated, terminal, terminal or intercalary in conidial chains, doliiform and pale brown; conidia which are branched chains, acrogenous, straight or slightly curved, dark brown to blackish, pale brown or subhyaline at apex, 1-3 septate, strongly constricted at the septa, verruculose or finely echinulate and rounded at both ends. A new species was introduced, based on morphological and phylogenetic analysis of combined ITS, LSU, RPB2 and TEF sequence data. Detailed descriptions and illustrations are provided, with an updated phylogenetic tree depicting intergeneric relationships within the Torulaceae.

Biodegradation of Gossypol by Aspergillus terreus-YJ01.

Gossypol, generally found in the roots, stems, leaves, and, especially, the seeds of cotton plants, is highly toxic to animals and humans, which inhibits the use of cotton stalks as a feed resource. Here, a promising fungal strain for biodegrading gossypol was successfully isolated from the soil of cotton stalk piles in Xinjiang Province, China, and identified as Aspergillus terreus-YJ01 with the analysis of ITS. Initial gossypol of 250 mg·L-1 could be removed by 97% within 96 h by YJ01, and initial gossypol of 150 mg·L-1 could also be catalyzed by 98% or 99% within 36 h by the intracellular or extracellular crude enzymes of YJ01. Sucrose and sodium nitrate were found to be the optimal carbon and nitrogen sources for the growth of YJ01, and the optimal initial pH and inoculum size for the growth of YJ01 were 6.0 and 1%, respectively. To further elucidate the mechanisms underlying gossypol biodegradation by YJ01, the draft genome of YJ01 was sequenced using Illumina HiSeq, which is 31,566,870 bp in length with a GC content of 52.27% and a total of 9737 genes. Eight genes and enzymes were predicted to be involved in gossypol biodegradation. Among them, phosphoglycerate kinase, citrate synthase, and other enzymes are related to the energy supply process. With sufficient energy, β-1, 4-endo-xylanase may achieve the purpose of biodegrading gossypol. The findings of this study provide valuable insights into both the basic research and the application of A. terreus-YJ01 in the biodegradation of gossypol in cotton stalks.

Morphological and phylogenetic analysis of Raillietina spp. in indigenous chickens (Gallus gallus domesticus) in Bangladesh

Raillietina spp. (Cestoda: Davaineidae), the most common cestodes in indigenous chickens, cause a substantial production loss in poultry industry in Bangladesh. Here, we estimated the prevalence, confirmed the species and determined the genetic pattern of species of Raillietina using molecular tools. We collected and examined 375 chickens randomly from household of different villages of Mymensingh sadar and Gouripur upazila, Mymensingh district and adult parasites were isolated and identified. Genomic DNA was extracted from collected parasites, amplified ITS-2 and ND-1 genes, sequenced and analyzed. Out of 375 samples, 270 (72.0%) were found positive with Raillietina species and mean worm burden was 10.46 ± 0.56. Microscopically, three species of Raillietina, such as R. cesticillus (37.9%), R. echinobothrida (41.1%) and R. tetragona (52.8%) were detected on the basis of their morphological features. The total length, length and width of scolex, sucker and rostellum were also measured. Among different factors, age, farming nature and flock size of chickens were significantly (p < 0.05) influenced Raillietina infections. For further validation, the sequences of ITS-2 gene generated in this study were matched with reference sequences of R. cesticillus, R. echinobothrida and R. tetragona and found 99.63% − 100% similarity. The phylogenetic analyses of ITS-2 and ND-1 sequences were clustered together with the reference sequences of R. cesticillus, R. echinobothrida and R. tetragona confirming microscopic identification. This is the first confirmation of species of Raillietina along with the prevalence of the species, which will be helpful for the formulation of a control strategy and provide basic information for further molecular study.

Monnina (Polygalaceae), a New World monophyletic genus full of contrasts

Abstract Endemic to the Neotropics, Monnina is the second largest genus of Polygalaceae, yet little is known about its phylogenetic history, biogeography, and morphological character evolution. To address these knowledge gaps, we conducted Bayesian and maximum likelihood (ML) analyses of nuclear ITS and plastid trnL–F regions to test the monophyly of Monnina s.l. We used this phylogenetic framework to (i) infer divergence time estimates of lineages within the genus and reconstruct their historical biogeography; (ii) reconstruct the evolution of morphological characters of putative ecological and evolutionary importance in Monnina; and (iii) test for correlations between our phylogenetic hypothesis and environmental data. Our results reveal that Monnina is monophyletic with an indehiscent, 1–2-seeded fruit as a synapomorphy for the genus. We identify six clades within Monnina based on our combined phylogenetic results: Clades A, B, and D are primarily distributed in southern and eastern South America, Clades C and E are primarily Central Andean, and Clade F is chiefly distributed in the Northern Andes and Central America. The ancestor of the Monnina stem lineage dispersed from Australia/Africa to South America during the late Eocene to early Oligocene. The divergences of major lineages within the genus began in the early Miocene. We inferred the most recent common ancestor of Monnina to be an herbaceous plant with one-seeded samaroid fruits. The origins of fleshy fruits and shrubby habits are phylogenetically correlated within Monnina, and their concerted convergent evolution may have promoted increased net diversification rates in the two most species-rich subclades of the genus.

Genetic diversity, phylogenetic and phylogeographic analysis of Anopheles culicifacies species complex using ITS2 and COI sequences.

Anopheles culicifacies is the major vector of malaria in Sri Lanka and the Indian subcontinent which is characterized as a species complex with five sibling species provisionally designated as A, B, C, D and E. The current study was carried out to understand the phylogenetic and phylogeographic relationships between the sibling species of the species complex while observing their genetic diversity and genetic differentiation. Thirty-five ITS2 and seventy-seven COI sequences of An. culicifacies species complex reported from different geographical locations of Asia and China at the NCBI public database were used for the analysis. Bayesian likelihood trees were generated for the phylogenetic analysis. The divergence of the species complex was obtained from the Bayesian phylogeographic model in BEAST. There were two clades of the sibling species of An. culicifacies species complex as A, D and B, C and E in both phylogenetic and phylogeographic analysis using ITS2 sequences. Based on the highly divergent COI sequences and the high mutation rate of the mitochondrial genome, there were four and three clades in both phylogenetic and phylogeographic analysis using COI sequences. The diversification of An. culicifacies species complex was obtained as ranging from 20.25 to 24.12 Mya and 22.37 to 26.22 Mya based on ITS2 and COI phylogeographic analysis respectively. There was a recent diversification of the sibling species A and D than the sibling species B, C and E. Low haplotype diversity was observed in the sequences reported from Sri Lanka in both ITS2 and COI analysis that can be due to bottlenecks resulting from the intense malaria control efforts. A high genetic differentiation was achieved for some populations due to the large geographical distance. The high genetic diversity based on the five sibling species implies the possibility of maintaining a relatively high effective population size despite the vector control efforts.

Which molecular marker is better? Comparative analyses of COI and ITS2 in molecular identification of Trichogramma (Hymenoptera: Trichogrammatidae)

Which molecular marker is better? Comparative analyses of COI and ITS2 in molecular identification of Trichogramma (Hymenoptera: Trichogrammatidae)

Comparison of assembly platforms for the assembly of the nuclear genome of Trichoderma harzianum strain PAR3

BackgroundTrichoderma is a diverse genus of fungi that includes several species that possess biotechnological and agricultural applications, including the biocontrol of pathogenic fungi and nematodes. The mitochondrial genome of a putative strain of Trichoderma harzianum called PAR3 was analyzed after isolation from the roots of Scarlet Royal grapevine scion grafted to Freedom rootstock, located in a grapevine vineyard in Parlier, CA, USA. Here, we report the sequencing, comparative assembly, and annotation of the nuclear genome of PAR3 and confirm its identification as a strain of T. harzianum. We subsequently compared the genes found in T. harzianum PAR3 to other known T. harzianum strains. Assembly of Illumina and/or Oxford Nanopore reads by the popular long-read assemblers, Flye and Canu, and the hybrid assemblers, SPAdes and MaSuRCA, was performed and the quality of the resulting assemblies were compared to ascertain which assembler generated the highest quality draft genome assembly.ResultsMaSuRCA produced the most complete and high-fidelity assembly yielding a nuclear genome of 40.7 Mb comprised of 112 scaffolds. Subsequent annotation of this assembly produced 12,074 gene models and 210 tRNAs. This included 221 genes that did not have equivalent genes in other T. harzainum strains. Phylogenetic analysis of ITS, rpb2, and tef1a sequences from PAR3 and established Trichoderma spp. showed that all three sequences from PAR3 possessed more than 99% identity to those of Trichoderma harzianum, confirming that PAR3 is an isolate of Trichoderma harzianum. We also found that comparison of gene models between T. harzianum PAR3 and other T. harzianum strains resulted in the identification of significant differences in gene type and number, with 221 unique genes identified in the PAR3 strain.ConclusionsThis study gives insight into the efficacy of several popular assembly platforms for assembly of fungal nuclear genomes, and found that the hybrid assembler, MaSuRCA, was the most effective program for genome assembly. The annotated draft nuclear genome and the identification of genes not found in other T. harzainum strains could be used to investigate the potential applications of T. harzianum PAR3 for biocontrol of grapevine fungal canker pathogens and as source of anti-microbial compounds.

Two novel species and three new records of Torulaceae from Yunnan Province, China.

While investigating the diversity of lignicolous fungi in Yunnan Province, China, six fresh collections of Torulaceae were collected and identified based on morphological examination and phylogenetic analyses of combined LSU, ITS, SSU, tef1-α, and rpb2 sequence data. Two new species, viz. Neopodoconisyunnanensis and Torulasuae, and three new records, viz. T.canangae (new freshwater habitat record), T.masonii (new host record), and T.sundara (new freshwater habitat record) are reported. Detailed descriptions, illustrations, and a phylogenetic tree to show the placement of these species are provided.

Dictyosporium duliujiangense sp. nov. (Dictyosporiaceae, Pleosporales) from freshwater habitat in Guizhou Province, China

A survey of the taxonomy and diversity of freshwater fungi from Karst Plateau wetlands in Guizhou Province, China, three fresh collections were collected from a freshwater river. They have cheiroid, digitate, dictyosporous conidia with one or two hyaline appendages. Phylogenetic analysis of the combined ITS, LSU and tef1-α sequence data support their placement with genera Dictyocheirospora and Dictyosporium (Dictyosporiaceae, Pleosporales). Two collections represent the same species, Dictyocheirospora rotunda. The other collection is introduced as a new species Dictyosporium duliujiangense. An updated phylogenetic backbone tree, descriptions and illustrations of the two taxa are provided for Dictyosporiaceae.

First Report of Corynespora cassiicola Causing Leaf Spot on Jasminum nudiflorum in China.

Winter jasmine (Jasminum nudiflorum Lindl.), a trailing, deciduous shrub, is widely used as an ornamental plant. Its flowers and leaves also has great medicinal value for treatment of inflammatory swelling, purulent eruptions, bruises and traumatic bleeding (Takenaka et al. 2002). In October 2022, leaf spot symptoms were observed on J. nudiflorum distributed in Meiling Scenic Spot (28.78°N, 115.83°E) and Jiangxi Agricultural University (28.75°N, 115.83°E), Nanchang, Jiangxi Province, China. In a week-long series of investigations, the incidences of disease could range up to 25%. Initially, the symptoms of the lesions were small yellow circular spots (0.5 to 1.8 mm), and gradually developing irregular spots (2.8 to 4.0 mm) with grayish white central parts, a dark brown inner ring, and outer yellow halo. To identify the pathogen, sixty symptomatic leaves from fifteen different plants were collected, of which twelve were randomly selected, cut into 4-mm2 pieces, and surface sterilized with 75% ethanol for 30s followed by 5% NaClO for 1 min, rinsed four times with sterile water, and then placed on potato dextrose agar (PDA) medium at 25 °C in the dark for 5 to 7 days. Six isolates with similar morphological characteristics were obtained. Aerial mycelium was vigorous, downy and exhibited white to grayish-green coloration. Conidia were solitary or catenate, pale brown, obclavate to cylindrical, apex obtuse, one to 11 pseudosepta, 24.9 to 125.7 × 7.9 to 12.9 μm (n = 50). Morphological characteristics matched Corynespora cassiicola (Ellis 1971). For molecular identification, two representative isolates (HJAUP C001 and HJAUP C002) were selected for genomic DNA extraction, and the ITS, TUB2 and TEF1-α gene were amplified, using the primer ITS4/ITS5 (White et al. 1990), Bt2a/Bt2b (Lousie and Donaldson 1995) and EF1-728F/EF-986R (Carbone and Kohn 1999), respectively. The sequenced loci (GenBank accession nos. ITS: OP957070, OP957065; TUB2: OP981639, OP981640; TEF1-α: OP981637, OP981638) of the isolates were 100, 99 and 98% similar to the corresponding sequences of C. cassiicola strains (GenBank accession nos. OP593304, MW961419, MW961421, respectively). Phylogenetic analyses of combined ITS and TEF1-α sequences was performed using maximum-likelihood method in MEGA 7.0 (Kuma et al. 2016). The result showed that our isolates (HJAUP C001 and HJAUP C002) clustered with four strains of C. cassiicola at 99% bootstrap values in the bootstrap test (1000 replicates). Based on the morpho-molecular approach, the isolates were identified as C. cassiicola. The pathogenicity of one representative strain (HJAUP C001) was tested by inoculating the wounded leaves of six healthy J. nudiflorum plants under natural condition. Three leaves from each of three plants were punctured with flamed needles and sprayed with a conidial suspension (1 × 106 conidia/ml), and three wounded leaves from each of other three plants were inoculated with mycelial plugs (5 × 5 mm3). Mock inoculations were used as controls with sterile water and PDA plugs on three leaves each, respectively. Leaves from all treatments were incubated in a greenhouse at high relative humidity, 25°C, and 12-hour photoperiod. After one week, all wounded inoculated leaves appeared similar symptoms as described above, whereas the mock inoculated leaves were still healthy. Similar isolates with grayish white and vigorous aerial mycelium were reisolated from inoculated and symptomatic leaves and identified as C. cassiicola by DNA sequencing, fulfilling Koch's postulates. It has been reported that C. cassiicola can cause leaf spots on numerous plant species (Tsai et al. 2015; Lu et al. 2019; Farr and Crossman 2023). However, to our knowledge, this is the first report of C. cassiicola causing leaf spots on J. nudiflorum in China. This finding aids in protection of J. nudiflorum, a medicinal and ornamental plant with high economic value.

Triplosphaeria guizhouensis sp. nov. (Tetraplosphaeriaceae, Pleosporales), a novel taxon from freshwater habitat in Guizhou Province, China

During our survey of the taxonomy and diversity of freshwater fungi of Guizhou Province, China, a Tetraploa-like taxon was collected from stream in China. The taxon has quite smaller, oblong, dark brown to black, three or four-columned conidia with three or four setose appendages. Phylogenetic analyses of combined LSU, SSU, ITS, tef1-α and β-tubulin sequence data revealed that the newly obtained isolate clustered as a monophyletic clade with the species of Triplosphaeria with strong statistical support (98% ML BS/1.0 PP). Based on the morphological comparison and multi-locus phylogeny, we introduce T. guizhouensis as a new species in Triplosphaeria. The description and illustration of the newly discovered taxon, and updated backbone tree of Tetraplosphaeriaceae are provided.

First report of Nigrospora sphaerica associated with leaf spot disease of Crossandra infundibuliformis in India.

Crossandra (Crossandra infundubuliformis (L.) Nees.) is one of the main floriculture crops in Karnataka. In 2020 (March-June), a characteristic leaf spot disease of unknown etiology with an incidence ranging from 10-12% (~30 ha area evaluated) was observed in Southern Karnataka (Mysore, Mandya). Initially, the symptoms developed as small specks (3 to 8 mm), characterized by circular to irregular shapes in the beginning and coalesced to form larger lesions. Ten samples were collected in polybags followed by the isolation of associated fungal pathogen on potato dextrose agar (PDA) medium amended with Chloramphenicol (60 mg/L). Briefly, small pieces of infected leaves were cut into small pieces and surface sterilized with 2% sodium hypochlorite (NaOCl) solution, rinsed three times with sterile distilled water (SDW), blot dried, then inoculated onto PDA medium, and incubated at room temperature (27 ± 2°C) for 3 - 5 days. Fungal colonies developed from the segments and were subcultured through hyphal tipping to fresh PDA plates to get pure cultures. A total of 12 pure cultures were obtained. Mycelia were initially white and eventually turned gray. The conidia were black, single-celled, smooth, spherical to subspherical, 9 to 18 μm in diameter (n=50), and borne singly on a hyaline vesicle at the tip of each conidiophore. The identity was initially established based on the cultural features and conidial morphology as Nigrospora sp. (Deepika et al., 2021). To confirm the identity of fungal isolates based on molecular sequence analysis was performed for two representative isolates (CIT1 & CIT2). ITS-rDNA, tub2 & EF-1α gene were amplified using primers ITS1/ITS4, T1/T22 & EF1-728F/986R (White et al., 1990; O'Donnel and Cigelnik, 1997; Carbone and Kohn, 1999), then purified and sequenced. The BLASTn analysis of ITS, tub2 and EF-1α gene showed 99-100% similarity with reference sequences from the GenBank database to Nigrospora sphaerica (ITS: 520bp, KX985935 - LC7312; MH854878 - CBS:166.26; tub2: 357bp, MZ032030 - WYR007, 350bp, KY019606 - LC7298, KY019522 - LC4278, KY019520 - LC4274; EF-1α: 472bp, KY019397 - LC7294, KY019331 - LC4241; MN864137 - HN-BH-3) and the sequences were deposited in GenBank (ITS: OL672271 & OL672272; tub2: OL782120 & OL782121; EF-1α: ON051604 & ON051605) (Wang et al., 2017). The associated fungal pathogen was identified as N. sphaerica (Sacc.) Mason (Chen et al. 2018; Deepika et al., 2021) based on the cultural, morphological, microscopic, and molecular characteristics. Further, pathogenicity tests were conducted on healthy plants (Crossandra cv. Arka; n=30) grown under greenhouse conditions (28±2 °C; 80% RH). Inoculations were made with conidial suspension (18 days old N. sphaerica isolate CIT1, 106 conidia/ml) prepared in SDW, and healthy plants sprayed with SDW (n=10) served as controls. All the plants were covered with polyethylene bags for 24-48 hr and observations were made at regular intervals. Typical necrotic lesions developed on 16 plants after 12 days after inoculation but no symptoms were observed on the control plants. The associated pathogen was re-isolated from diseased leaves and confirmed their identity based on morphology and cultural characteristics. Earlier, N. sphaerica was associated with various tree species as an endophyte, and recently several reports have appeared to cause disease on various crop plants (Deepika et al., 2021). However, there are no previous reports on the association of N. sphaerica causing leaf spot disease on C. infundibuliformis from India. Early diagnosis of this leaf spot disease will help the floriculturist adopt suitable management practices to avoid significant economic loss.

Limnofasciculus baicalensis gen. et sp. nov. (Coleofasciculaceae, Coleofasciculales): A New Genus of Cyanobacteria Isolated from Sponge Fouling in Lake Baikal, Russia.

The proliferation of benthic cyanobacteria has been observed in Lake Baikal since 2011 and is a vivid manifestation of the ecological crisis occurring in the littoral zone. The cyanobacterium Symplocastrum sp. has formed massive fouling on all types of benthic substrates, including endemic Baikal sponges. The strain BBK-W-15 (=IPPAS B-2062T), which was isolated from sponge fouling in 2015, was used for further taxonomic determination. A polyphasic approach revealed that it is a cryptic taxon of cyanobacteria. Morphological evaluation of the strain indicated the presence of cylindrical filaments with isodiametric cells enclosed in individual sheaths and coleodesmoid false branching. Strain ultrastructure (fascicular thylakoids and type C cell division) is characteristic of the Microcoleaceae and Coleofasciculaceae families. An integrated analysis that included 16S rRNA gene phylogeny, conserved protein phylogeny and whole-genome comparisons indicated the unique position of BBK-W-15, thus supporting the proposed delineation of the new genus Limnofasciculus. Through characterisation by morphology, 16S, ITS and genomic analysis, a new cyanobacterium of the family Coleofasciculaceae Limnofasciculus baicalensis gen. et sp. nov. was described.

A new species of the genus Anamylopsora (Baeomycetaceae; Ascomycota) from Deosai National Park, Gilgit-Baltistan, Pakistan

AbstractA novel lichen species occurring on rocks was collected from three different localities within Deosai National Park, Gilgit-Baltistan, Pakistan. Phylogenetic analyses of the nrDNA ITS and nuLSU regions revealed that it clustered within the genus Anamylopsora. Further chemical and morpho-anatomical analyses confirmed its uniqueness, and it is described here as a new species under the name A. pakistanica. The distinguishing characters are: an irregularly squamulose appressed thallus on rocks without rhizines; an epinecral layer up to 25 μm thick; ascospores that are hyaline, simple, thick-walled with a smooth surface; septate paraphyses with a pigmented apical cell in a gel-like matrix; globose to subglobose pycnidia with hyaline and bacilliform pycnidiospores. In particular, the species is distinguished from other members of the genus by morpho-anatomical features including the coloration of the thalli, the presence of a thick lower cortex (up to 100 μm), and the presence of simple, thick-walled ascospores. Specimens were found at altitudes up to 4587 m, the highest elevation yet reported for Anamylopsora. A key and comparison to all existing species of the genus Anamylopsora is also given.

Taxonomy, phylogeny and evolution of freshwater Hypocreomycetidae (Sordariomycetes)

Hypocreomycetidae is a highly diverse group with species from various habitats. This subclass has been reported as pathogenic, endophytic, parasitic, saprobic, fungicolous, lichenicolous, algicolous, coprophilous and insect fungi from aquatic and terrestrial habitats. In this study, we focused on freshwater fungi of Hypocreomycetidae which resulted 41 fresh collections from China and Thailand. Based on morphological and phylogenetic analyses, we identified 26 species that belong to two orders (Hypocreales and Microascales) and six families (Bionectriaceae, Halosphaeriaceae, Microascaceae, Nectriaceae, Sarocladiaceae and Stachybotryaceae). Ten new species are introduced and 13 new habitats and geographic records are reported. Mariannaea superimposita, Stachybotrys chartarum and S. chlorohalonatus are recollected from freshwater habitats in China. Based on phylogenetic analysis of combined LSU, ITS, SSU, rpb2 and tef1-α sequences data, Emericellopsis is transferred to Hypocreales genera incertae sedis; Pseudoacremonium is transferred to Bionectriaceae; Sedecimiella is placed in Nectriaceae; Nautosphaeria and Tubakiella are excluded from Halosphaeriaceae and placed in Microascales genera incertae sedis; and Faurelina is excluded from Hypocreomycetidae. Varicosporella is placed under Atractium as a synonym of Atractium. In addition, phylogenetic analysis and divergence time estimates showed that Ascocodina, Campylospora, Cornuvesica and Xenodactylariaceae form distinct lineages in Hypocreomycetidae and they evolved in the family/order time frame. Hence, a new order (Xenodactylariales) and three new families (Ascocodinaceae, Campylosporaceae and Cornuvesicaceae) are introduced based on phylogenetic analysis, divergence time estimations and morphological characters. Ancestral character state analysis is performed for different habitats of Hypocreomycetidae including freshwater, marine and terrestrial taxa. The result indicates that marine and freshwater fungi evolved independently from terrestrial ancestors. The results further support those early diverging clades of this subclass, mostly comprising terrestrial taxa and freshwater and marine taxa have been secondarily derived, while the crown clade (Nectriaceae) is represented in all three habitats. The evolution of various morphological adaptations towards their habitual changes are also discussed.

Current status and characterization of Phytophthora species associated with gummosis of citrus in Northern Vietnam

AbstractPhytophthora species have long been reported to cause Phytophthora disease in citrus with symptoms of gummosis, root rot and fruit brown rot. Due to the lack of an up‐to‐date investigation of Phytophthora species currently causing the disease in Northern Vietnam, this study aimed to determine the current Phytophthora species associated with the disease in this region. An extensive collection of diseased samples was conducted in the main citrus‐growing areas in Northern Vietnam from 2021 to 2022. Fifty‐four Phytophthora isolates were obtained and initially divided into four groups based on morphological characteristics, including colony morphology, mycelial growth rate and reproductive structures. Representative Phytophthora isolates from each group were then identified as P. palmivora, P. nicotianae, P. citrophthora and P. mekongensis based on the phylogenetic analysis of ITS and cox1 gene sequences. Pathogenicity tests showed that these four Phytophthora species induce gummosis and brown rot on pomelo, orange and lime, with pomelo being the most susceptible, followed by orange and lime, respectively. Koch's postulates were confirmed. All the Phytophthora species were sensitive to various fungicides including mancozeb, metalaxyl, chlorothalonil, fosetyl aluminium, potassium phosphonate and dimethomorph, of which metalaxyl was the most potent inhibitor of colony growth. Our findings provide updated information on the current Phytophthora species associated with gummosis in citrus in Northern Vietnam, with P. mekongensis being recorded for the first time in this region. This will serve as a reference for Phytophthora studies on citrus in the future and for developing an integrated management strategy for Phytophthora disease.

Varicosporellopsis shangrilaensis sp. nov. (Nectriaceae, Hypocreales), a new terricolous species isolated from the rhizosphere soil of Astragalus polycladus in northwestern Yunnan, China

Two fungal strains were isolated from the rhizosphere soil of a Chinese medicinal herb plant, Astragalus polycladus, in northwestern Yunnan, China. Both of them were identified as a new fungal species of Varicosporellopsis based on the evidence of morphology and phylogenetic analysis of ITS, LSU, TUB and ACT sequences data. Phylogenetic analysis of combined ITS and LSU sequence data revealed that strains MTF 7 and KLF 01 belong to the genus Varicosporellopsis with the highest sequence similarity (95.51%) to the type strain V. americana CBS 148257 and formed a sister lineage to V. americana CPC 40768 with high support values (100% ML, 1.00 BI). Morphologically, V. shangrilaensis is characterized by macronematous, branched conidiophores, phialidic conidiogenous cells, reniform macroconidia and spherical microconidia. This new species is the first reported of Varicosporellopsis in China. The morphological description, illustrations and phylogenetic analysis of V. shangrilaensis are provided in this study.

A new species of Calocybe (Agaricales, Basidiomycota) from China

Morphological and molecular phylogenetic data were used to describe a new Calocybe species (C. violacea) collected from Yunnan Province, China. Calocybe violacea is characterized by its convex to applanate, purple to bluish-purple pileus; purple-black, bluish-purple to white stipe; and relatively larger basidiospores. Phylogenetic analyses of the combined ITS and nrLSU loci supported the distinct position of this proposed new species, showing that C. violacea clustered in a single sister clade with C. vinacea.