Phylogenetic Evidence Research Articles

Morphology, phylogeny, and mitogenomics reveal a new entomopathogenic fungus, Blackwellomyces changningensis (Hypocreales, Clavicipitaceae), from southwestern China

ABSTRACT Two species of Blackwellomyces (Clavicipitaceae, Hypocreales, Ascomycota) were discovered during an investigation of the diversity of entomopathogenic fungi. A new fungus and one known fungal species that were gathered from Yunnan Province were described in this study. Blackwellomyces changningensis, sp. nov. was described using morphology and phylogenetic evidence from 14 mitochondrial protein-coding gene (PCG) data sets (atp6, atp8, atp9, cob, cox1, cox2, cox3, nad1, nad2, nad3, nad4, nad5, nad6, and nad4L) and six nuclear genes (ITS [ITS4 and ITS5], 18S nuc rDNA [18S], 28S nuc rDNA [28S], tef1-α, rpb1, and rpb2). B. changningensis were found parasitic to Lepidoptera larvae and to produce filiform ascospores with septations. The asexual conidia were ovoid to ellipsoid in shape. Phylogenetic analysis and morphological observations concurred that the fungus belonged to a different species within the genus of Blackwellomyces. The foundation for further taxonomic, genetic, and evolutionary biological studies of the genus Blackwellomyces was laid by this work.

Mallotus bullatus (Euphorbiaceae), a new species from Southwest China based on morphological characters and phylogenetic evidence

Mallotus bullatus M.T.An & J.H.Yu, sp. nov. (Euphorbiaceae), a species new to science discovered in Guizhou, China, is described and illustrated here, and its phylogenetic position among other Mallotus species is presented. Morphological, micro-morphological, and molecular evidence is presented as attestation of its novelty. The new species morphologically resembles M. philippensis var. reticulatus and M. philippensis var. philippensis, but it clearly differs by having bullate leaf surfaces (vs. not bullate), leaf margins entire or nearly so (vs. entire or nearly so in M. philippensis var. philippensis and coarsely serrate in M. philippensis var. reticulatus), leaf margins sometimes bearing red glands (vs. red glands absent), 5 sepals in staminate flowers (vs. 3–4 in M. philippensis var. philippensis and 4 in M. philippensis var. reticulatus), fruits with spines (vs. spines absent in M. philippensis var. philippensis and present in M. philippensis var. reticulatus), and abaxial leaf epidermal scattered and clustered vein hairs 0.1–0.8 mm long (vs. 0.04–0.28 mm long in M. philippensis var. philippensis and 0.05–0.1 mm long in M. philippensis var. reticulatus). Molecular phylogenetic analysis (BS = 100% / BS = 96%, PP = 1 / PP = 1) provides strong evidence supporting M. bullatus as a new species within the genus Mallotus and supports its placement in M. sect. Philippinenses as sister to M. philippensis.

Sarea cirrhendocarpa, a fungus species new to science from the southern Rocky Mountains

Sarea cirrhendocarpa, a resinicolous fungus, is here described as new to science from two specimens collected in the Indian Peaks Wilderness (Arapaho-Roosevelt National Forest, Colorado, USA) during the summer of 2023, a third specimen from Holy Cross Wilderness (White River National Forest, Colorado, USA) during the summer of 2024, and a fourth specimen from the Kachina Peaks Wilderness (Coconino National Forest, Arizona, USA) that was collected in 1998. The new species is readily distinguished from its closest relative, Sarea resinae, by its larger apothecia, smaller asci, smaller ascospores, and burnt orange hymenium (bicolored towards uppermost portions) among other traits. A phylogeny derived from the nuclear ribosomal internal transcribed spacer (ITS) region suggests that S. cirrhendocarpa is sister to and genetically distinct specimens of Sarea resinae sampled from around the world including North America. These molecular results in combination with the clear morphological differences lead us to propose this as a new species so far only known from the southern Rocky Mountains of North America. This new taxon is placed for the time being within the genus Sarea owing to inconclusive phylogenetic and morphological evidence that would support generic separation within the Sareomycetes. The name Sarea resinae is retained and new combinations for S. klamathica and S. lignicola are provided.

Orphan genes are not a distinct biological entity.

The genome sequencing revolution has revealed that all species possess a large number of unique genes critical for trait variation, adaptation, and evolutionary innovation. One widely used approach to identify such genes consists of detecting protein-coding sequences with no homology in other genomes, termed orphan genes. These genes have been extensively studied, under the assumption that they represent valid proxies for species-specific genes. Here, we critically evaluate taxonomic, phylogenetic, and sequence evolution evidence showing that orphan genes belong to a range of evolutionary ages and thus cannot be assigned to a single lineage. Furthermore, we show that the processes generating orphan genes are substantially more diverse than generally thought and include horizontal gene transfer, transposable element domestication, and overprinting. Thus, orphan genes represent a heterogeneous collection of genes rather than a single biological entity, making them unsuitable as a subject for meaningful investigation of gene evolution and phenotypic innovation.

Four New Species of Deconica (Strophariaceae, Agaricales) from Subtropical Regions of China

Deconica is a relatively small genus, with only 90 names recorded in previous research. In this study, four new species of Deconica have been identified based on morphological and phylogenetic evidence from subtropical regions of China. This represents the first discovery of new species of Deconica in China. Morphologically, D. austrosinensis is characterized by medium-sized spores that are elliptical to elongated-ellipsoid in face view, and fusiform to sublageniform and slightly thick-walled pleurocystidia; D. furfuracea is identified by a well-developed and evanescent veil, medium-sized spores that are rhomboid to mitriform in face view, and fusiform to subclavate pleurocystidia that are rare and subacute at apex; D. fuscobrunnea is recognized by dark brown pileus, medium-sized spores that are rhomboid to mitriform in face view, an ixocutis pileipellis, lageniform cheilocystidia with a long neck and lacks pleurocystidia; D. ovispora is distinguished from other Deconica species by medium-sized spores that are ovoid in face view, an ixocutis pileipellis, lageniform cheilocystidia with a long to short neck, and lacks pleurocystidia. Their distinct taxonomic status is confirmed by the positions of the four new species in ITS + LSU phylogenetic trees. Detailed descriptions and morphological photographs of four new species are presented.

First Report of Alternaria alternata Causing Leaf Spot of Magnolia delavayi in China

Magnolia delavayi, known as Delavay's magnolia or Chinese evergreen magnolia, is an enchanting deciduous tree native to southwestern China (Xu et al. 2020). In March 2024, leaf spots were observed on M. delavayi plants at Qujing Normal University (25.527°N, 103.744°E), Qujing, Yunnan, China. About 80% of the trees (n=92) on campus were infected, with infected leaves accounting for about 10 to 30% of all leaves. The leaf spots exhibited a round or oval shape, with a yellowish-brown center and a brown outer ring. As the spots expanded, the leaves gradually dried up and fell off. Five symptomatic leaves were collected from the middle sections of five diseased plants. Tissue samples (5 × 5 mm) were excised from the lesion margins and subjected to surface sterilization in 75% ethanol for 30 seconds and 1% sodium hypochlorite for 60 seconds. They were then rinsed three times with sterile water and dried on sterilized paper. The samples were plated on potato dextrose agar (PDA) and incubated at 28 °C for 3 days. Developing colonies were retransferred to new PDA and purified by the hyphal tip technique (Senanayake et al. 2020). Two representative isolates (SYL1 and SYL2) were selected and confirmed to be the same species based on morphological characteristics and molecular identification. On PDA, the fungal colonies initially appeared grayish-white and turned dark gray over time with cotton-like aerial hyphae on the surface. The conidia are light brown, pear-shaped or rod-shaped, with 1 to 4 transverse septa and 0 to 1 oblique septa, and measured 5.4 to 12.7 × 10.4 to 25.7 μm (n = 50). Molecular identification was performed by partial sequencing of the internal transcribed spacer (ITS: ITS4/ITS5) region, glyceraldehyde 3-phosphate dehydrogenase (GAPDH:gpd1/gpd2), translation elongation factor 1-alpha (TEF1:EF1/EF2), and Alternaria major allergen (Alt a 1:Alt-for/Alt-rev) (Woudenberg et al. 2015). The resulting sequences of ITS (PP951882, PP951883), GAPDH (PP968942, PP968943), Alt a 1(PP968940, PP968941) and TEF1 (PP968938, PP968939) were deposited in GenBank. BLAST analyses revealed that the sequences from these two isolates showed 100% identity with those from Alternaria alternata strains for each gene. Additionally, a multigene phylogenetic analysis revealed a distinct group containing SYL1, SYL2 and other A. alternata strains with a posterior probability of 100%. The morphological and phylogenetic evidence collectively suggested that the two isolates belonged to A. alternata. Strain SYL1 was used for the pathogenicity test on three 5-year-old M. delavayi plants on campus. Three healthy leaves of each plant were punctured with a sterile needle and sprayed with 20 μl of spore suspension (1 × 106 conidia/ml). Similarly, another nine wounded leaves were sprayed with 20 μl of sterile water. All inoculated leaves were enclosed in plastic bags for 48 hours. Disease symptoms were observed on leaves inoculated with spore suspension after 7 days, while control plants remained healthy. This experiment was repeated three times with same results. To fulfill Koch’s postulates, A. alternata was reisolated from the inoculated leaves and identified based on morphological characteristics and ITS sequencing. To our knowledge, this is the first report of A. alternata causing leaf spot on M. delavayi in China. This research expands the known host range of this pathogen and provides relevant information to the design of disease management strategies.

Morpho-phylogenetic evidence reveals Pseudobionectria phoenicis gen. et sp. nov. (Bionectriaceae) from a mangrove habitat

A bionectriaceous ascomycete was collected from palm rachides in a mangrove habitat. Multi-gene phylogenetic analyses indicated this fungus belongs within the Bionectriaceae, forming a sister lineage to Septofusidium herbarum and distinct from other known genera. In this study, we proposed the new genus Pseudobionectria to accommodate the novel species P. phoenicis. Pseudobionectria is distinguished from other genera in Bionectriaceae by its superficial, globose, brownish-orange ascomata with a black crown that is flattened on the top and hyphal appendages that merge with the subiculum below. The asci are unitunicate, lacking a visible apical ring, and the ascospores are 1-septate, oblong-ellipsoidal to fusiform, hyaline, and exhibit spinose ornamentation. The establishment of this new genus is supported by morphological and phylogenetic evidence. Additionally, this paper provides a synopsis of the genera within Bionectriaceae, including information on the type species, morphological characteristics, lifestyles, molecular data and key publications.

Leaf Rust Pathogens on Hypericum pseudohenryi: Describing Melampsora danbaensis sp. nov. and M. hyperici-pseudohenryi sp. nov. from China.

Based on morphological and phylogenetic evidence, two novel species of Melampsora were discovered on Hypericum pseudohenryi in China and have been thoroughly characterized. One of these species, designated as M. danbaensis, exhibits distinct features such as aecia of Uredo-type, typically appearing in gregarious or grouped arrangements, and presenting a shallowly pulvinate structure. Aeciospores exhibit tremendous variations in size, ranging in shape from globose to ellipsoidal and bearing pronounced verrucose texture. Telia resemble crusts one-spore deep, covering nearly the entire abaxial leaf surface, with sessile teliospores reaching sizes of up to 65.8 µm, and exhibiting a clavate to cylindrical shape. Another species, designated as M. hyperici-pseudohenryi, is distinguished by Uredo-type uredinia, which are hypophyllous, scattered or grouped, and interspersed with numerous paraphyses. Its urediniospores tend to be globose, ellipsoidal or obovoid, echinulate, and are accompanied by clavate to capitate paraphyses reaching lengths up to 77.6 µm. Phylogenetically, both species form a novel monophyletic clade within the Melampsora genus, with robust support demonstrated by a high Maximum likelihood bootstrap support (MLBS) value of 100% and a Bayesian posterior probability (BPP) of 1. This study enriches our understanding of the diversity and geographical distribution of Melampsora species that infect Hypericum plants in China.

Linking intercontinental biogeographic events to decipher how European vineyards escaped Pierce's disease.

Global change is believed to be a major driver of the emergence of invasive pathogens. Yet, there are few documented examples that illustrate the processes that hinder or trigger their geographic spread. Here, we present phylogenetic, epidemiological and historical evidence to explain how European vineyards escaped Xylella fastidiosa (Xf), the vector-borne bacterium responsible for Pierce's disease (PD). Using Bayesian temporal reconstruction, we show that the export of American grapevines to France as rootstocks to combat phylloxera (~1872-1895) preceded the spread of the Xf grapevine lineage in the USA. We found that the time of the most recent common ancestor in California dates to around 1875, which agrees with the emergence of the first PD outbreak and the expansion into the southeastern US around 1895. We also show that between 1870 and 1990, climatic conditions in continental Europe were mostly below the threshold for the development of PD epidemics. However, our model indicates an inadvertent expansion of risk in southern Europe since the 1990s, which is accelerating with global warming. Our temporal approach identifies the biogeographical conditions that have so far prevented PD in southern European wine-producing areas and predicts that disease risk will increase substantially with increasing temperatures.

Aspergillus capibaribensis (Eurotiales, Trichocomaceae), a new species in section Cremei

In this study we describe a new species of Aspergillus in section Cremei deposited in the URM culture collection (Micoteca URM Profa. Maria Auxiliadora Cavalcanti) of the Departamento de Micologia, Universidade Federal de Pernambuco (Recife, Pernambuco, Brazil). Another strain was obtained from samples taken from pollen pots inside nests of Melipona scutellaris (stingless bee). Morphological and phylogenetic evidence based on sequencing of ITS rDNA, BenA, CaM and rpb2 markers supported the description of taxonomic novelty. Aspergillus capibaribensis sp. nov. is morphologically similar and phylogenetically close to A. flaschentraegeri - the distinctive features are uniseriate and hyaline conidiophores, short and smooth conidia and smaller vesicles compared to the micromorphological structures of A. flaschentraegeri. This record also expanded the knowledge of species in Aspergillus section Cremei in a tropical country such as Brazil.

Microbial communities and metagenomes in methane-rich deep coastal sediments

Coastal sediments are rich in embedded recalcitrant organic carbons that are biotransformed into methane. In this study, gas composition (carbon dioxide, methane and nitrogen) and chemical indicators (total nitrogen, total carbon, and total sulfate) were examined in five deep sediment cores (up to 130 m in length) obtained from the Hangzhou Bay. The V3-V4 region of the 16S rRNA gene amplicons was amplified and sequenced for the prokaryotic community analysis. The species composition, along with the physicochemical factors of the sediments, revealed a strong correlation with methane content in one of the sediment cores. We then obtained metagenomes of the two sediment samples selected for their high methane content and enrichment of methanogenic Bathyarchaeota with phylogenetic evidence. A total of 27 draft genomes were retrieved through metagenomic binning methodologies and were classified into Bathyarchaeota, Asgard archaea, Planctomycetes, and other microbial groups. The data provided are valuable for understanding the relationship between methane generation and microbial community composition in deep sediment core samples from coastal to marine environments.

The Combination of Morphological and Phylogenetic Evidence Reveals Four New Gymnopus Species and New Distribution.

The genus Gymnopus plays a significant role in ecological systems, with certain species holding potential as food or medicinal resources. However, the species diversity of Gymnopus in China remains unclear. In recent years, more than one thousand Gymnopus specimens have been collected across China. Thus, through the integration of ecological evidence, detailed morphological studies, and phylogenetic analysis using a multiloci dataset of ITS + nLSU + tef1-ɑ, four new species-Gymnopus longistipes, Gymnopus striatipileatus, Gymnopus viridiscus, and Gymnopus spadiceus-have been differentiated from known species. Gymnopus similis has been newly documented from Jiangxi Province, China. Detailed descriptions and vivid illustrations have been provided based on the newly collected specimens, along with comparisons to closely related species. Additionally, a key to the reported species of Gymnopus s.l. from East China has been included.

Phylogeography and species distribution modeling unveil unnoticed Pliocene diversity: the case of a montane Iberian bush-cricket, Antaxius spinibrachius (Orthoptera: Tettigoniidae)

Abstract The geographic distribution and systematics of many groups of insects are still understudied across large geographic areas of the Iberian Peninsula. This lack of knowledge and the fact that many species have a complex evolutionary history due to the existence of “refugia within refugia” have hindered the taxonomic description of the true biodiversity in the Iberian Peninsula. Here, we discuss the evolutionary history of a flightless Iberian bush-cricket Antaxius spinibrachius (Fischer, 1853) using 1 nuclear and 2 mitochondrial markers. We applied species distribution modeling to design sampling strategies in climatically favorable areas and to detect missing isolated populations or unknown lineages. Following the species distribution modeled using all of the presence records available, we discovered 4 new isolated populations in Sierra Madrona, Serra de São Mamede, Sierra de Montánchez, and Sierra de la Demanda. Phylogenetic analyses recovered 2 major Pliocene lineages with a north–south geographic speciation pattern in the Iberian Peninsula. This north–south vicariant event split the common ancestor of these species on either side of the Tagus Valley, probably associated with the Pliocene climate change and the reorganization of paleobasins of the western Iberian Peninsula during the Miocene–Pliocene. We described the southern Iberian Plateau lineage of Antaxius as a new species of bush-cricket based on phylogenetic and morphological evidence, Antaxius oretanus sp. nov. We assessed the conservation status of the new species as “VU B2ab(iii,v)” under the IUCN criteria.

Morphological and molecular identification of two new Marasmiellus species (Omphalotaceae, Agaricales) from Thailand.

Marasmiellus (Omphalotaceae, Agaricales) specimens collected in Thailand were investigated based on morphological characteristics and molecular phylogenetic analyses. In the present study, two species are introduced as new to science, namely Marasmiellusthailandicus and M.minutisporus. Phylogenetic analyses were carried out based on the internal transcribed spacer (nrITS) and nuclear ribosomal RNA large subunit (nrLSU) regions, and the results revealed that the two new taxa are distinct species within Marasmiellus. Another specimen was identified as M.scandens and is reported for the first time with morphology and molecular data from Thailand. Descriptions, illustrations, and phylogenetic results are provided. In addition, M.diaphanus and M.colocasiae are proposed as new combinations of Collybiopsisdiaphana and Paramarasmiuscolocasiae, respectively, based on the phylogenetic evidence.

Phylogenomic and molecular marker based studies to clarify the evolutionary relationships amongst Anoxybacillus species and demarcation of the family Anoxybacillaceae and some of its constituent genera.

The family Anoxybacillaceae was recently proposed encompassing the genera Anoxybacillus, Geobacillus, Parageobacillus, Saccharococcus and Thermolongibacillus. Of these genera, Anoxybacillus contains >50% of the Anoxybacillaceae species. However, Anoxybacillus species form multiple unrelated clades in phylogenetic trees and their evolutionary relationships are unclear. To clarify the evolutionary relationships of Anoxybacillus and other Anoxybacillaceae species, detailed phylogenomic and comparative analyses were conducted on 38 Anoxybacillaceae species with available genomes. In a phylogenomic tree based on 1148 core proteins, all Anoxybacillus, Geobacillus, Parageobacillus, Saccharococcus and Thermolongibacillus species, excepting Anoxybacillus sediminis, formed a strongly supported clade representing the family Anoxybacillaceae. Five conserved signature indels (CSIs) reported here are also uniquely found in these species, providing robust means for the demarcation of family Anoxybacillaceae in molecular terms. In our phylogenomic tree and in the Genomic Taxonomy Database, Anoxybacillus species formed four distinct clades designated as Anoxybacillus sensu stricto (containing the type species A. pushchinoensis), Anoxybacillus_A, Anoxybacillus_B and Anoxybacillus_C. Our analyses have identified 17 novel CSIs which offer means to reliably distinguish species from these clades based upon multiple uniquely shared molecular characteristics. Additionally, we have identified three and seven CSIs specific for the genera Geobacillus and Brevibacillus, respectively. All seven Brevibacillus-specific CSIs are also shared by Anoxybacillus sediminis, which branches reliably with this genus. Based on the strong phylogenetic and molecular evidence presented here, we are proposing that the genus Anoxybacillus should be restricted to only the species from Anoxybacillus sensu stricto clade, whereas the species from Anoxybacillus_A, Anoxybacillus_B, and Anoxybacillus_C clades should be transferred into three novel genera Anoxybacteroides gen. nov., Paranoxybacillus gen. nov. and Thermaerobacillus gen. nov., respectively. Additionally, we are also proposing the transfer of Anoxybacillus sediminis to the genus Brevibacillus. The proposed changes, which reliably depict the evolutionary relationships among Anoxybacillaceae species, should be helpful in the studies of these organisms.

Phylogenetic and morphological evidence for four new species of Russula (Russulaceae, Basidiomycota) from northwestern China

Four novel species of subgenus Russula crown clade collected from northwestern China are described based on morphological and phylogenetic evidence. Morphologically, R. griseorosea Y.Song sp. nov. (subsection Puellarinae) is characterized by its brown pileus with a grayish pink tint, basidiospores with warts often connected by fine lines, orthochromatic pileipellis with long terminal cells and septate pileocystidia; R. micangshanensis Y.Song sp. nov. (subsection Olivaceinae) is diagnosed by its large basidia, hymenial cystidia and basidiospores, and spore ornamentations with unequal crests and often twinned warts, which give the spore distinctive appearance; R. minirosea Y.Song sp. nov. (subsection Laricinae) has very small basidiocarp with pileus less than 3.3 cm in diameter, basidiospores with fine reticulum, small basidia, and septate pileocystidia; R. purpureomarginalis F.Li & Y.Song sp. nov. (subsection Xerampelinae) has large basidiospores with often isolated ornamentations, slim basidia and often septate flexuous pileocystidia. Differences between the four novel species and their closely related taxa were analyzed. Phylogenetic analyses based on both ITS and multi-locus (LSU, rpb2 and tef1) were carried out to confirm the distinct taxonomic status of the four novel species.

Phylogenetic evidence of extensive spatial mixing of diverse HIV-1 group M lineages within Cameroon but not between its neighbours.

From the perspective of developing relevant interventions for treating HIV and controlling its spread, it is particularly important to comprehensively understand the underlying diversity of the virus, especially in countries where the virus has been present and evolving since the cross-species transmission event that triggered the global pandemic. Here, we generate and phylogenetically analyse sequences derived from the gag-protease (2010 bp; n = 115), partial integrase (345 bp; n = 36), and nef (719 bp; n = 321) genes of HIV-1 group M (HIV-1M) isolates sampled between 2000 and 2022 from two cosmopolitan cities and 40 remote villages of Cameroon. While 52.4% of all sequenced viruses belonged to circulating recombinant form (CRF) 02_AG (CRF02_AG), the remainder were highly diverse, collectively representing seven subtypes and sub-subtypes, eight CRFs, and 36 highly divergent lineages that fall outside the established HIV-1M classification. Additionally, in 77 samples for which at least two genes were typed, 31% of the studied viruses apparently had fragments from viruses belonging to different clades. Furthermore, we found that the distribution of HIV-1M populations is similar between different regions of Cameroon. In contrast, HIV-1M demographics in Cameroon differ significantly from those in its neighbouring countries in the Congo Basin (CB). In phylogenetic trees, viral sequences cluster according to the countries where they were sampled, suggesting that while there are minimal geographical or social barriers to viral dissemination throughout Cameroon, there is strongly impeded dispersal of HIV-1M lineages between Cameroon and other locations of the CB. This suggests that the apparent stability of highly diverse Cameroonian HIV-1M populations may be attributable to the extensive mixing of human populations within the country and the concomitant trans-national movements of major lineages with very similar degrees of fitness; coupled with the relatively infrequent inter-national transmission of these lineages from neighbouring countries in the CB.

Proposal of Lactobacillus amylovorus subsp. animalis subsp. nov. and an emended description of Lactobacillus amylovorus.

Seven novel lactic acid bacterial strains (BF125T, BF186, TKL145, YK3, YK6, YK10 and NSK) were isolated from the fresh faeces of Japanese black beef cattle and weanling piglets, spent mushroom substrates, or steeping water of a corn starch production plant. These strains are rod-shaped, Gram-stain-positive, non-motile, non-spore-forming, catalase-negative, cytochrome oxidase-negative, facultatively anaerobic, and homofermentative. Strain BF125T did not produce any gas from glucose; both d- and l-lactate were produced as end-products of glucose (D/L, 40 : 60). Growth occurred at 30-45 °C (optimum, 37 °C), pH 5.0-8.0 (optimum, pH 6.0), and with NaCl concentration of 1.0-3.0% (w/v). The G+C content of genomic DNA of strain BF125T was 37.8 mol% (whole-genome analysis). The major fatty acids were C16 : 0, C18 : 1 ω9c, C19 cyclopropane 9, 10, and summed feature 10. The 16S rRNA gene in strain BF125T showed high similarity to that of the type strain of Lactobacillus amylovorus (99.93%), and the other isolates were also identified as L. amylovorus based on these similarities. A phylogenetic tree based on the core genomes of L. amylovorus strains (n=54), including the seven isolates, showed that they could be divided into two clusters. Strains YK3, YK6, YK10, and NSK were in the first cluster, along with the type strain DSM 20531T, while the second cluster included isolates BF125T, BF186, TKL145, and other strains isolated from various animal origins. Phenotypic differences in fermentability were observed for lactose, salicin, and gentiobiose between these two groups. The intergroup digital DNA-DNA hybridization values (72.9-78.6%) and intergroup average nucleotide identity values (95.64-96.92%) were comparable to values calculated using datasets of other valid subspecies of the genus (ex-) Lactobacillus. In light of the physiological, genotypic, and phylogenetic evidence, we propose a novel subspecies of L. amylovorus, named Lactobacillus amylovorus subsp. animalis subsp. nov. (type strain BF125T=MAFF 212522T=DSM 115528T). Our findings also led to the automatic creation of Lactobacillus amylovorus subsp. amylovorus subsp. nov. and an emended description of the species L. amylovorus.

Four novel species of Pleurotheciaceae collected from freshwater habitats in Jiangxi Province, China.

During an investigation of fungal diversity from freshwater environments in different regions in Jiangxi Province, China, four interesting species were collected. Morphology coupled with combined gene analysis of an ITS, LSU, SSU, and rpb2 DNA sequence data showed that they belong to the family Pleurotheciaceae. Four new species, Pleurotheciella ganzhouensis, Pla. irregularis, Pla. verrucosa, and Pleurothecium jiangxiense are herein described. Pleurotheciella ganzhouensis is characterized by its capsule-shaped conidia and short conidiophores, while Pla. irregularis has amorphous conidiophores and 3-septate conidia. Pleurotheciella verrucosa has cylindrical or verrucolose conidiogenous cells, 1-septate, narrowly fusiform, meniscus or subclavate conidia. Pleurothecium jiangxiense characterized in having conidiogenous cells with dense cylindrical denticles and short conidiophores. Pleurothecium obovoideum was transferred to Neomonodictys based on phylogenetic evidence. All species are compared with other similar species and comprehensive descriptions, micrographs, and phylogenetic data are provided.

Seven New Species of Entoloma Subgenus Cubospora (Entolomataceae, Agaricales) from Subtropical Regions of China.

Entoloma is a relatively large genus in Agaricales, with a rich diversity of species and a wide distribution. In this study, seven new species of Entoloma belonging to the subgenus Cubospora have been identified based on morphological and phylogenetic evidence from subtropical regions of China. Morphologically, E. excavatum is characterized by the yellow, depressed, estriate pileus and medium-sized basidiospores; E. lacticolor is recognized by the white and papillate pileus, adnexed lamellae, and presence of clamp connections; E. phlebophyllum is identified by the pink-to-maroon and estriate pileus, and lamellae with lateral veins; E. rufomarginatum differs from other cuboid-spored species by the lamellae edge which is red-brown-underlined; E. subcycneum is characterized by the white pileus and carneogriseum-type cheilocystidia; E. submurrayi is recognized by the pileus margin exceeding the lamellae, 2-layered pileipellis with hyphae of different widths, and the presence of clamp connections; E. tomentosum is identified by the tomentose pileus, heterogeneous lamella edge, and versiform cheilocystidia with brown-yellow contents. Their distinct taxonomic status is confirmed by the positions of the seven new species in both the ITS + LSU and 3-locus (LSU, tef-1α, rpb2) phylogenetic trees. Detailed descriptions, color photos, and a key to related species are presented.