Gagea kotuchovii (Liliaceae) a new narrow endemic species growing exclusively in the Karatau Mountains in South Kazakhstan, is here described and illustrated. It is a unique species within the section Gagea, differing from closely related taxa by the presence of several stolons of different lengths (0.5–8 cm), formed on a single vegetative individual, as well as a unique bulb with a densely woolly sheath. Morphological characteristics, distribution map, and illustrations of the habit and habitats of the new species are presented. We also present phylogenetic analyses based on the internal transcribed spacer (ITS) of nuclear DNA and SNPs obtained from DArT genome-wide sequencing (Diversity Arrays Technology sequencing), which confirmed the isolated position of G. kotuchovii but also revealed its phylogenetic relation with morphologically similar species. Additionally, our results reveal phylogenetic intergeneric organization of Gagea representatives occurring in Kazakhstan and/or Middle Asia. To facilitate morphological identification of Asian Gagea with stolons, and similar to G. kotuchovii, we present an identification key.

Carnation and Dianthus (Dianthus spp) are globally cultivated as a cut flower, yet high summer temperatures in Taiwan significantly reduce the yield and quality of commercial cultivars. To enhance stress tolerance traits such as heat resistance and disease resistance, interspecific and intergeneric hybridization with native species is a viable approach. Since the success of hybridization is influenced by genetic distance, this study aimed to clarify the phylogenetic relationships among native species, commercial cultivars, and interspecific hybrids using internal transcribed spacer (ITS) and random amplified polymorphic DNA (RAPD) markers. Phylogenetic analysis based on RAPD markers effectively differentiated among native Taiwanese species, commercial varieties, interspecific hybrids, and outgroup taxa. ITS markers, on the other hand, were more informative for identifying parent-offspring relationships. Notably, Dianthus superbus var. longicalycinus from Taiwan and Japan, despite sharing the same scientific name, exhibited clear distinctions in both morphological traits and molecular profiles. The combined use of RAPD and ITS markers provides complementary insights into the genetic relationships within the Dianthus genus. These findings not only support the strategic use of molecular markers in breeding programs but also highlight the need to reassess taxonomic classifications among morphologically similar varieties. This study provides genetics, breeding tools, and germplasm information for future Dianthus breeding.

Here we describe two new species of Inocybe from pine forests of Pakistan; I. hazarensis and I. shimlaensis . Morphological and molecular data show that these species have not been described before and hence need to be described as new. Both species are smooth‐spored and pruinose only in the apical part of the stipe. Inocybe hazarensis is characterized by rather small size, brown to dark brown, dense radial fibrils to rimose to glabrous pileus with prominent umbo, finely fibrillose at apex only becoming pruinose to strigose, along rest of length of stipe, slightly bulbous stipe base, narrow basidiospore spores and smaller clavate, oblong, ovoid, narrowly utriform cheilocystidia. Inocybe shimlaensis is characterized by brown to golden pileus, low and broad umbo, radially fibrillose, rimose to granulose, with brown appressed‐squamulose stipe with only apex pruinose and submarginate base. Anatomically, it has small (6.8 × 4.4 μm) basidiospores and smaller conical to fusiform cheilocystidia. Phylogenetic estimation based on DNA sequences from the internal transcribed spacer (ITS) region and large subunit (LSU) of the nuclear ribosomal DNA (rDNA) genes is congruent with the morphological characters that help to delimit these as new species of Inocybe .

Deciphering microbial diversity and predicting metabolic functionalities in fermented pigmented rice water using culture-independent characterization.

Acer saccharum Marsh., valued for its ornamental, material, and edible uses, is an important temperate tree species in the Northern Hemisphere. A blight disease affecting branches of A. saccharum was first identified in 2023 in Shandong, China. The causal agent was identified as Botryosphaeria wangensis G.Q. Li & S.F. Chen based on cultural and morphometric characteristics. Phylogenetic analysis was performed by amplifying and sequencing the internal transcribed spacer (ITS) region of rDNA, the translation elongation factor 1α (tef1) partial gene, the β-tubulin (tub2) partial gene, and the second largest subunit of RNA polymerase II (rpb2), in combination with morphological data. Symptoms observed in the field were replicated in a pathogenicity test through inoculation of A. saccharum branches, thus satisfying Koch’s postulates. To our knowledge, this is the first report worldwide of B. wangensis infecting A. saccharum.

The perennial Arabis paniculata is a member of the Brassicaceae widely used as a model species for perennial flowering (Wang et al. 2009). In April 2023, typical powdery mildew symptoms were observed on A. paniculata at Longwen Hill (26°23' N, 106°37' E), Guizhou Normal University, China. Disease incidence reached ~100% among 60 wild A. paniculata plants examined. Mycelia appeared sporadically distributed on leaf surfaces. Hyphal appressoria were lobed, solitary or in opposite pairs. Hyphae measured 5.5-7.5 μm wide, exhibiting branched, straight to flexuous morphology. Conidiophores were erect, straight to slightly flexuous, and 50-125 µm long (n = 30). Foot cells were followed by 2 or 3 cells. Conidia formed singly, ellipsoid, occasionally in chains of two, and measured 30-45 μm long × 14-18 µm wide when shed (n = 50). Based on these morphological characteristics, the powdery mildew fungus (designated GZAA-1) was provisionally identified as an Erysiphe species (Braun and Cook 2012). To confirm the species, the internal transcribed spacer (ITS) and large subunit (LSU) regions were respectively amplified using primer pairs ITS1/ITS4 (White et al. 1990) and NL1/NL4 (Ziemiecki et al. 1990). The obtained 649-bp ITS sequence (GenBank accession no. PV682595) showed 100% similarity (649/649-bp) to previously reported ITS sequences (such as AB522714 and OQ195144) of E. cruciferarum. The obtained 617-bp LSU sequence (PV682641) had 100% similarity (616/616-bp) with LSU sequences (LC009943 and OQ195144) of E. cruciferarum. Sequencing of a second isolate (GZAA-2) confirmed 100% ITS and LSU sequence identity with GZAA-1. Phylogenetic analysis of concatenated ITS (649-bp) and LSU (616-bp) sequences further clustered GZAA-1 within the E. cruciferarum clade alongside reference strains. Pathogenicity tests were conducted by spray-inoculating three healthy potted A. paniculata plants with a suspension of fresh conidia (1 × 10⁵ spores/mL). Three control plants were treated with distilled water. All plants were maintained in a greenhouse at 25 ± 2°C and 80% relative humidity. Typical powdery mildew symptoms developed on inoculated plants after 12-15 days, whereas controls remained asymptomatic. The pathogen reisolated from inoculated plants was morphologically identical to the original field isolate. These tests were repeated three times with consistent results. Collectively, morphological, molecular, and pathogenicity data confirmed the pathogen as E. cruciferarum. While E. cruciferarum has been reported to infect A. paniculata in Romania (Braun 1995), this study represents the first report of E. cruciferarum-induced powdery mildew on A. paniculata in China. These findings expand the known geographical distribution of E. cruciferarum on this host.

The soil fungus Penicillium gladioli was identified by sequencing of the internal transcribed spacer (ITS) region. Gas chromatography-mass spectroscopy of the extract showed 50 compounds. Among the detected peaks, n-hexadecanoic acid showed the largest relative peak area (7.989% of the total ion chromatogram), followed by phenol, 2-methyl-5-(1-methylethyl) (6.543%). Regarding the in vivo anti-toxoplasma potential, there was an enhancement of the histological features of the liver of Swiss albino mice with a substantial decrease (p < 0.05) in the inflammatory mediators, including cyclooxygenase-2, tumor necrosis factor-alpha, interleukin-6, and interleukin-1β. The colorimetric determination of the nitric oxide and malondialdehyde in the liver of the fungal extract-treated group revealed its antioxidant effect by significantly reducing (p < 0.05) the oxidative stress markers. P. gladioli extract established antibacterial potential on P. aeruginosa bacteria with a minimum inhibitory concentration of 64 to 512µg/ml. Moreover, it demonstrated antibiofilm potential using crystal violet assay and SEM. Also, 45% of the isolates displayed downregulation of the lasR, lecA, and pelA biofilm genes.

Cladosporium tongheensis sp. nov. was isolated from diseased Allium ochotense leaves displaying characteristic symptoms collected in Heilongjiang Province, China. Morphologically, the novel species exhibits branched, septate hyphae ranging in color from subhyaline and pale brown to distinctly brown, with surfaces smooth or slightly roughened. Conidiophores arise singly, erect, occasionally curved, and originate either laterally or terminally from hyphae. They are septate, smooth to faintly rough, pale brown to brown in color, occasionally branched, and display notable constrictions and swellings at septal junctions. The conidia are abundant, mostly obovoid, limoniform, or ellipsoid, predominantly aseptate, occasionally possessing one septum, smooth-surfaced, and pale to medium brown. Secondary ramoconidia exhibit shapes varying from cylindrical and subcylindrical to oblong-ellipsoid and oblong. Their color ranges from pale to distinctly brown, containing between zero and four septa, and typically tapering at the base with pronounced constrictions occurring at the septa. Multi-gene phylogenetic analysis, involving actin (act), ribosomal internal transcribed spacer (ITS) regions and translation elongation factor-1α (tef1), unequivocally positioned these isolates as a novel phylogenetic lineage with close affinity to Cladosporium basiinflatum, situated within the broader Cladosporium herbarum species complex. Therefore, combining phylogenetic evidence and morphological data, we propose a novel species named C. tongheensis.

Echeveria plants are a large genus of succulents belonging to the Crassulaceae family. Echeveria plants are suitable for urban living and are therefore highly popular in Japan. Since 2015, symptoms of disease have been observed on E. laui and E. agavoides cultivar Evony in commercial greenhouses (34°57'58.3"N, 136°28'37.6"E) in Mie Prefecture, Japan. First, water-soaked symptoms appeared at the leaf base, the plant then softened, and the leaves dropped off, or abundant gray mold and conidia were formed on the rotted plants. Approximately 5–6% of the plants were affected, which appeared indiscriminately on species or hybrids and was more prevalent from late March to early July. The fungus was isolated from the cultivars Blood Maria × Lindsayana and Monroe Chanel. The affected leaves were surface-sterilized with 1% sodium hypochlorite for 10 s, followed by 70% ethanol for 60 s, rinsed three times with sterile water, and excised into 10 mm 2 squares. Samples were placed on a Botrytis selective medium (Edwards and Seddon 2001). After subculturing the fungal colonies on potato dextrose agar (PDA), 16 isolates were obtained through single-conidial isolation. Fungal colonies of all isolates on PDA were cottony flocculent texture with a color ranging from gray-white to light brown. Strain Ec23_1 was selected as a representative isolate for further studies. The conidia were colorless or gray, oval, and elliptical, measuring 9.0–19.1 × 6.3–15.8 μm (n=131), and were produced in gapelike clusters on the terminal end of the branch conidiophore. Many small sclerotia (0.8–2.3 × 0.7–1.6 mm, n=76) were irregularly produced on PDA at a later stage of growth. The morphology of the isolate closely resembled that of Botrytis (Ellis 1971). For molecular analysis, genomic DNA was extracted using a commercial DNA extraction kit (Qiagen, Hilden, Germany). Nuclear protein-coding genes of glyceraldehyde 3-phosphate dehydrogenase (G3PDH), heat shock protein 60 (HSP60), RNA polymerase II (RPB2), and internal transcribed spacer (ITS) were amplified and sequenced using the specific primers (Staats et al. 2005). All sequences were deposited in GenBank under accession numbers LC890466 (ITS), LC890460 (G3PDH), LC890461 (HSP60), and LC890462 (RPB2). BLAST analyses revealed that the ITS, G3PDH, HSP60, and RPB2 sequences exhibited 99–100% (562/562, 922/925, 1030/1030, 1129/1134 bp) identity to B. cinerea (GenBank: OQ257130, PV470194, MN159921, and MN448501, respectively). Based on morphological and molecular features, the isolate Ec23_1 was identified as B. cinerea. To confirm pathogenicity, young plants of the Sarang cultivar were sprayed with conidial suspensions (10 5 conidia ml -1 , n=9). The inoculated plants were placed in a plastic box and maintained at 24°C under a 12-h photoperiod with high humidity (&gt;90%) overnight, followed by 40%. Symptoms appeared on the lower leaves within 1 week after inoculation, and the plants were finally covered with gray mold. The re-isolated fungus showed the same morphological features and pathogenicity as the original isolates, fulfilling Koch’s postulates. In Crassulaceae, gray mold or rot diseases have been reported on Sedum sarmentosum in Korea caused by B. cinerea and E. gigantea in Mexico by Sclerotinia sclerotiorum. To our knowledge, this is the first report of B. cinerea on Echeveria plants in Japan. This study provides useful information on plant disease management to maintain stable seedling sales and value in the Echeveria market.

Amorphophallus bulbifer (Roxb.) Blume is a perennial tuber that belongs to the family Araceae, is widely used in food, pharmaceutical, and chemical industries because of its richness in glucomannan. It is mainly distributed in southern Yunnan of China, northern Myanmar, Laos, northern Thailand and tropical rain forest areas in Indonesia (Gao et al, 2022). In July to August 2023, a leaf blight disease occurred on A. bulbifer in Ankang Academy of Agricultural Sciences manufacturing base (32°69′N, 109°02′E), Shaanxi, China. About 100 plants were investigated, and the results showed that the incidence of leaf blight disease was 36%. The symptoms were characterized by yellow-brown and irregular lesions. With time, the lesions enlarged and affected leaves finally became wilted. To investigate the disease, ten plants with typical symptoms were collected, one leaf was selected from each plant, and cut into 2 mm × 2 mm pieces. Their surfaces were disinfected with 1.5% NaClO for 60s followed by 75% ethanol for 30s, rinsed three times with sterile distilled water, and finally incubated on 1.5% potato dextrose agar (PDA) plates at 28℃. A total of six isolates were obtained through single-spore cultivation. The colonies on PDA were fluffy with abundant aerial mycelia and covered the whole plates (90 mm in diameter) in seven days. Fungal colonies were white, then turned to grayish with an unevenly distributed (Fig.1f-g). Conidia were single-celled, black, smooth, spherical to subspherical, 15-18 μm in diameter (Fig.1h-j). These morphological characteristics were similar to those of Nigrospora sphaerica (Wang et al.2017). For molecular confirmation, genomic DNA was extracted from mycelia of the pathogen using an Ezup column fungal genomic DNA extraction kit (Sangon Biotech, Shanghai, China). To confirm the identity of the pathogen, the genomic fragments for the internal transcribed spacer (ITS), TEF-1 and TUB2 gene of the isolate were amplified by PCR (Duan et al., 2024) and sent for sequencing. The resultant sequence (GeneBank ID of gene ITS, TEF-1, TUB2 are PQ330916, PQ778340, PQ778339, respectively) were compared with the voucher specimens. BLAST results showed &gt;99% identity with those of Nigrospora sphaerica (GeneBank ID of Nigrospora sphaerica strain LC7298 ITS, TEF-1, TUB2 are KX985937, KY019401, KY019606, respectively). A neighbor joining phylogenetic tree with the concatenated sequences of these genes showed that Ab-B23 had the closest match with Nigrospora sphaerica (Fig. 2). Based on morphological characteristics and molecular dataset analyses, the isolates were identified as Nigrospora sphaerica. For pathogenicity test, twenty healthy 1-year-old Amorphophallus bulbifer plants were disinfected with 75% alcohol before inoculation, then sprayed with conidial suspensions (100 µL, 1×10⁶ conidia/ml sterile water) of the isolates; the other twenty plants, also disinfected with 75% alcohol before inoculation, were sprayed with sterile water as the controls. All the treated plants were cultivated in a glasshouse at 28±2℃ and 80 ± 5% relative humidity. Seven days after inoculation, a yellow-brown spot appeared on leaf surfaces, and the spot gradually expanded. Fifteen days after inoculation, infected leaves showed yellow-brown and irregular lesions, whereas the control plants remained symptomless (Fig.1a-e). The pathogen was reisolated from infected leaves with an 85% reisolation percentage, and was confirmed as Nigrospora sphaerica by morphology and molecular identification. To our knowledge, this is the first report of leaf blight disease of Amorphophallus bulbifer caused by Nigrospora sphaerica in Shaanxi Province of China. Our report would be helpful to Amorphophallus bulbifer growers to recognize this leaf blight disease, and corresponding measures could be taken to minimize or avoid the economic losses caused by the disease.

A multi-marker sequencing approach was applied to characterize the eukaryotic microbial community of the Najafgarh Drain. It will provide the first comprehensive view of eukaryotic diversity in this system, highlighting both its ecological significance and pathogenic potential. We combined 18S rRNA, internal transcribed spacer (ITS), and whole-metagenome shotgun (WMS) sequencing; 18S rRNA captured broad protist diversity but offered limited fungal resolution, which was complemented by ITS, while WMS provided higher, often species level resolution across kingdoms. To improve WMS classification, a dual-database strategy was employed, sequences were first classified using a eukaryote-filtered NCBI clustered nr database, unclassified sequences were subsequently analyzed with a combined database comprising of SILVA, MIDORI2, PlantITS, and COInr. This approach consistently increased the number of identified genera and species compared with using a single database. Using the multi-marker and dual database approach, multiple phyla were detected, including Chlorophyta (55.72%), Ciliophora (21.46%), Ascomycota (6.44%), Mucoromycota (1.35%), and Nematoda (0.80%). Beneficial taxa such as Chlorella, members of Ciliophora, and Penicillium, known for organic-matter degradation, nutrient cycling, and heavy-metal remediation, were observed. Additionally, human pathogens such as Pneumocystis jirovecii, Rhizopus arrhizus and Trichuris trichiura along with plant pathogenic fungi such as Fusarium, Sporisorium, and Rhizopus were also observed. These findings underscore the need to incorporate eukaryotic pathogen surveillance into water quality monitoring and environmental policy frameworks. Such measures would contribute to more resilient water management practices and support broader public health protection goals.

The genus Gomphus Pers. presents persistent taxonomic challenges due to its morphological similarities with related genera. In this study, we collected two specimens of Gomphus from Guizhou, China; one specimen is described as a new species, Gomphus bijiensis sp. nov. and the other is identified as G. matijun J.W. Liu & F.Q. Yu based on morphological traits and phylogenetic analyses of the nuclear rDNA internal transcribed spacer (ITS) and nuclear rDNA large subunit (LSU). To resolve their evolutionary relationships we assembled and annotated the mitochondrial genomes of both species using next-generation sequencing. Comparative analyses revealed codon usage strongly biased toward A- or U-ending codons, consistent with the low GC content typical of fungal mitochondria. Variation in protein-coding gene lengths and base composition suggests that diverse evolutionary pressures have shaped these genomes. Divergence time estimation indicates that morphological diversity within Gomphus and related macrofungi has largely resulted from convergent evolution. Phylogenetic reconstruction places G. bijiensis and G. matijun within a distinct clade, supporting their close evolutionary affinity and the coexistence of ancestral and derived traits. This study provides the first comprehensive mitochondrial genomic data for Gomphus, offering new insights into its taxonomy, phylogeny, and evolutionary dynamics, and establishing a framework for future studies within the Gomphaceae.

Powdery mildew disease is one of the major fungal diseases, affecting most of the crops including stone fruits. Due to their economical significance, the precise identification proof of the causal fungus is fundamental for their effective control. Seventy-five powdery mildew samples were collected across Kashmir valley (J&K, India) from each of the three stone fruits viz., peach (Prunus persica), plum (Prunus domestica) and apricot (Prunus armeniaca) for the taxonomic studies of associated fungi. The powdery mildew fungi were identified based on morphological features and by molecular characterization using ribosomal DNA internal transcribed spacer (ITS) region. Three species from two genera viz., Podosphaera and Erysiphe of Erysiphacea family were identified. The species identified were P. pannosa on peach, P. ampla on apricot and Erysiphe prunastri on plum. Amongst the identified fungal species, P. ampla on apricot is hitherto unreported from Asia, P. pannosa on peach and E. prunastri on plum from India.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-23119-3.

Mulberry (Morus spp.) has been cultivated for silkworm rearing for thousands of years in Asia. In February 2023, a bud blight disease was observed in Wuhan, Hubei Province, China (30°25′37″N, 113°57′39″E), affecting 12% of 2-year-old ‘YSD10’ mulberry trees. Symptoms included brown blight around winter buds with dense orange clusters, bud death due to necrosis, and progressive branch dieback from coalescing lesions. Symptomatic tissues (5× 5 mm) were excised from lesions margins, surface-sterilized (75% ethanol for 30 s, 2.5% sodium hypochlorite for 2 min), rinsed thrice with sterile water, and plated on potato dextrose agar (PDA). Three isolates (Cd5, Cd10, and Cd11) were single-spore purified. On PDA at 25°C for 7 days, they formed sparse white mycelia with pale/reddish-orange pigment, growing 4.25–4.79 mm day -1 . Microconidia were scarce, fusiform, oval, or obovoid. Macroconidia were fusiform, with slightly curved apical cells and foot-shaped basal cells, with 1-5 septa: 1-septate (12.7–30.4 × 2.4–3.6, avg. 22.3 × 3.0 μm), 3-septate (29.7–42.6 × 2.7–3.7, avg. 35.3 × 3.2 μm), 5-septate (39.2–53.2 × 3.2–4.5, avg. 47.3 × 3.7 μm) (n =30). Mesoconidiophores emerged laterally from aerial hyphae, simple or branched. Chlamydospores were globose to subglobose, intercalary, solitary or in chains, 3.8–6.0 μm in diameter (avg. 4.6 μm, n = 30). For molecular identification, ribosomal internal transcribed spacers (ITS), translation elongation factor 1α (EF-1α), RNA polymerase II largest subunit (RPB1), and second largest subunit (RPB2) were amplified using primers ITS1/ITS4 (White et al. 1990), EF1H/EF2T (O’Donnell et al. 1998), Fa/G2R (O'Donnell et al. 2010), and fRPB2-7cF/RPB2-11aR (Liu et al. 1999), respectively. Sequences were deposited in GenBank (ITS: PQ034582-PQ034584, EF-1α, RPB1, RPB2: PQ014390-PQ014398). Maximum-likelihood phylogenetic analysis (MEGA7) showed the three strains formed a distinct clade outside known Fusarium tricinctum species complex (FTSC), with 100% bootstrap support. Based on morphological and molecular characteristics (Crous et al. 2021), they are propose as a new species, Fusarium moricola sp. nov. Koch’s postulates were fulfilled using healthy 35-day-old mulberry seedlings and one-year-old ‘YSD10’ winter stems. For seedling inoculation, each strain was tested on three seedlings. Needle-wounded plants were sprayed with10 7 conidia ml -1 (5 ml/plant), maintained humidity at 25 °C under an 18/6 h light/dark. For stems inoculation, buds of surface sterilization stems were wounded, ~ 1 cm of epidermis below each bud was excised, and each bud was wrapped in 5-layer sterilized toilet paper saturated with 3 ml of 10 5 conidia ml -1 , then incubated at 25°C in darkness. Each strain was tested on three stems (5 buds/stem). Controls were treated with sterile water. At 7 days post-inoculation (dpi), brown lesions appeared on inoculated seedling nodes. At 14 dpi, inoculated stems developed orange sporodochia and brown discoloration matching field symptoms. Controls were asymptomatic. Fungi with identical morphology to the original isolates were reisolated from symptomatic tissues of both tests. Mulberry bud (twig) blight has been attributed to Fusarium lateritium f.sp. mori (Vajna 2000). To our knowledge, this is the first report of a novel Fusarium species within FTSC causing mulberry bud blight. The identification of this pathogen provides a basis for developing effective control strategies.

Guava root-knot nematode (Meloidogyne enterolobii Yang &amp; Eisenback, 1983) is a highly polyphagous Meloidogyne species that causes significant damage to several economically important crops worldwide. The expansion of M. enterolobii is increasing with recent reports emerging from the Middle East and North Africa regions (Ibrahim et al. 2023). Its strains are highly pathogenic and have the capacity to break various sources of resistance against root-knot nematodes (Vashisth et al. 2024; Yang and Eisenback 1983). Due to its relevance, M. enterolobii has been recognized as an A2 pest by the European and Mediterranean Plant Protection Organization (EPPO 2021). In September 2023, tomato plants (Solanum lycopersicum L. cv. Castlerock) with stunting and yellowing symptoms were observed in open fields in both Najran (17°32'10.2"N 44°14'27.0" E) and Jazzan (17°23'50.1"N 42°32'55.0" E) regions, Saudi Arabia. Upon uprooting of these diseased plants, galled roots were observed. Soil and galled roots were taken to the nematology lab of King Saud University for proper diagnosis. Nematodes were extracted from soil using the modified sugar flotation method (Jenkins 1964) and from infected roots using the protocol described by Hussey and Barker (1973). Nematode population densities ranged from 2,200 to 2,500 individuals per 250 g of soil, while 9,864 and 11,388 eggs and second-stage juveniles (J2s) were recovered per root system in Najran and Jazzan, respectively. Nematodes were characterized morphologically and molecularly using DNA sequence analysis of the 28S D2D3 (Subbotin et al., 2006) and the internal transcribed spacer (ITS) (Subbotin et al. 2001) regions of the ribosomal DNA. The females were pearly white, globular, and had an annulated surface. The perineal pattern was typical of M. enterolobii, with an oval shape and moderately high dorsal arch. The average size of adult females (n = 32); was (L: 610 ± 51 µm; W: 435 ± 35 µm), stylet length (15.3 ± 1.4 µm), excretory pore to anterior end and stylet length ratio (4.3 ± 0.3 µm). The J2s presented a thin, bluntly pointed tail with a clearly defined hyaline terminus. The average J2 size (n = 32) body length was (472 ± 28 µm), tail length (41.6 ± 8.3 µm), and stylet length (13.8 ± 0.4 µm). The observations of the M. enterolobii populations from Saudi Arabia were consistent with those previously described (Subbotin et al. 2021; Yang and Eisenback 1983). For molecular characterization, juveniles hatched from a single egg mass were used for DNA extraction. DNA sequences were deposited in GenBank under accession numbers PV267741 (D2D3), PV267743 (D2D3), PV605585 (ITS), and PV605584 (ITS). The D2D3 sequences showed 99.86 % identity with M. enterolobii sequences from India (OP935640) and China (MT193449), while ITS sequences showed 98.28 - 99.86 % identity to M. enterolobii sequences from China (MT028373), India (MK940246), and Mexico (OM721664). To fulfill Koch’s postulates, 2-week-old seedlings of tomato cv. Marmande were transplanted into 25 cm-diameter pots containing 1.5 kg sterilized sandy loam soil and peat moss (3:1 v/v). A total of 7 pots were used. Each pot contained one tomato seedling and was infested with 3,000 J2s and eggs. Non-inoculated plants served as controls. After 62 days, infected plants showed root galls. No symptoms were observed in the controls. Adult females from galled roots were reidentified based on the rDNA regions as M. enterolobii. To our knowledge, this is the first report of M. enterolobii in Saudi Arabia.

Malassezia species are lipophilic yeasts that inhabit the skin of warm-blooded animals and that are associated with various skin disorders. Although Malassezia is frequently isolated from the external ear canal of cats, the influence of ectoparasites such as Otodectes cynotis (ear mites) on Malassezia species diversity has received limited attention. During an investigation of Malassezia diversity in cat ear canals infested with Otodectes cynotis, five Malassezia strains were isolated from the external ear canals. Phylogenetic analyses based on the internal transcribed spacer (ITS) region and the D1/D2 domains of the Large Subunit rDNA (LSU rDNA) revealed that those five isolates represent two known species, namely Malassezia globosa and Malassezia slooffiae, and a putative novel candidate species of Malassezia. The candidate species was found to be closely related to Malassezia gallinae and M. slooffiae, yet it differed from M. gallinae by 78 nucleotides (nt) in the ITS region and 9nt in the D1/D2 domains, and from M. slooffiae by 70nt in the ITS region and 5nt in the D1/D2 domains. Based on phylogenetic analysis and phenotypic characteristics, we propose a novel species for which we suggest the name Malassezia cafarchiae sp. nov.

Purpureocorticium niveum sp. nov., discovered in Yunnan Province, southern China, is described here as a new species based on morphology and phylogenetic analyses. Purpureocorticium niveum is characterized by its cracking basidiomata with a smooth hymenial surface, a monomitic hyphal system with clamp connections on generative hyphae, and pyriform basidiospores. Phylogenetic analyses of the new species were conducted based on internal transcribed spacer (ITS) and large subunit (LSU) sequences. The phylogenetic analyses revealed that the new species belongs to the genus Purpureocorticium and was recovered as a sister species to P. microsporum. However, morphologically, P. microsporum differs from P. niveum by having pinkish purple basidiomata and relatively smaller basidiospores. A full description, illustrations, and phylogenetic analyses results of the new species are provided in this article.

The mola carplet (Amblypharyngodon mola) is a well-adored small indigenous fish species (SIS) in Bangladesh. To evaluate the molecular and classical identification of the fungus associated with dried fish, the mola carplet was the subject of the current study. Aspergillus lentulus, a very common fungus in mola carplet was characterized molecularly using a 590 bp PCR product of its ITS (internal transcribed spacer) region. The molecular phylogenetic tree constructed therefrom showed 100% sequence similarity in A. lentulus species complex. The impact of cultural medium on mycelial growth and development of A. lentulus was pointed. The mycelial growth of A. lentulus was highest (71 mm) on the potato dextrose agar (PDA) medium, followed by (53 mm) in glucose peptone (GLP), (51 mm) in potato sucrose agar (PSA), (39 mm) in yeast extract agar (YEA), and (29 mm) Hoppkins (29 mm), respectively. A. lentulus grew and developed vegetatively best at 30°C and 7.0 pH, respectively. As far as we are aware, this experimental identification of the dry fish fungus Aspergillus lentulus from the mola carplet is the first report and confirmation of a fungus in Bangladesh using both traditional and molecular methods. Bangladesh J. Zool. 53(2): 209-218, 2025

Despite their significance, the diversity of acetic acid bacteria (AAB) in homemade vinegars remains understudied. This study aimed to explore the species-level diversity of AAB in homemade vinegars and to assess their community structure to better understand this microbial niche. To investigate the diversity of these bacteria, we employed recently established customized 16S-23S rDNA internal transcribed spacer (ITS) amplicon metagenomics to identify AAB at the species level. By applying Hill numbers, we calculated species richness, relative abundance, and dominance, providing a clearer understanding of the community structure of AAB in 11 homemade vinegars. Although species richness was relatively high, both relative abundance and dominance were considerably lower, suggesting a community structure dominated by a few highly abundant taxa, with most species being rare or low in abundance. The most dominant genera across most samples were Komagataeibacter and Acetobacter, both of which are known for their roles in oxidative fermentation. Several previously unreported, potentially novel species of AAB were identified, along with two potentially novel genera. This is one of the first studies to examine the diversity of AAB in homemade vinegars using a culture-independent amplicon metagenomic approach. Our findings suggest that the microbiota of homemade vinegars remains an underexplored niche and a source for novel species with biotechnological potential. The results provide valuable baseline data for future microbial studies and may help in the development of novel, customized starter cultures for the improvement and standardization of vinegar production.

Walnut (Juglans regia L.), an economically important woody oil crop, is widely cultivated in southern Xinjiang. In August 2024, field surveys conducted across five walnut orchards (covering approximately 20 hectares) in Kalatal Town, Aksu Prefecture, Xinjiang (80.4933°E, 40.7665°N) revealed that 30-70% of trees exhibited leaf necrosis symptoms. The initial symptoms appeared as small, irregular brown necrotic lesions along leaf margins, which progressively expanded across the entire leaf surface as the disease developed. A total of 12 symptomatic leaves were collected from three walnut orchards. Leaf segments (4×4 mm) and twig sections (10 mm in length) were excised from lesion margins, surface-sterilized sequentially in 75% ethanol for 30 s and 1% sodium hypochlorite for 3 min, rinsed 7-8 times with sterile distilled water, dried with sterile filter paper towels, and placed on potato dextrose agar (PDA) for 5-7 d at 28°C. Eight single-spore strains were obtained using the single-spore isolation method. After 4 days of incubation, the colonies initially appeared white to light pink, gradually developing a distinct pink pigmentation upon maturation. The macroconidia produced by this strain were falcate or elliptical, measuring 4.6-16.6 µm× 1.6-4.0 µm (n = 50). Genomic DNA was extracted from the representative isolate KY11 using the cetyltrimethylammonium bromide (CTAB) protocol. Four genetic loci were amplified and sequenced: the elongation factor 1-α (EF-1α) gene, internal transcribed spacer (ITS) region, 28S large ribosomal subunit (LSU) rRNA gene, and calmodulin (CAL) gene, along with the second largest subunit of RNA polymerase II (RPB2) gene. Amplification was performed using the following primer pairs: EF1-α: EF1-728F/EF1-986R, ITS: ITS1/ITS4, LSU: LR0R/LR5, CAL: CL1/CL2A, RPB2: fRPB2-5F/fRPB2-7Cr (White et al. 1990; Schoch et al. 2012; Xia et al. 2019). The sequences of isolate KY11 were deposited in GenBank with the following accession numbers: ITS(PV441516), EF-1α(PV524932), LSU(PV441478), CAL (PV536123), RPB2 (PV524934). BLASTn analysis revealed high sequence similarity with F. incarnatum references: 99.8% (ITS: MN871572.1), 99.61% (EF-1α: ON292430.1), 99.22% (LSU: MG274307.1), 100% (CAL: GQ505503.1), and 98.61% (RPB2: MT889350.1). A maximum likelihood phylogenetic tree was constructed based on concatenated sequences of EF-1α, CAL, and RPB2 using RAxML 8.0.0 software. Phylogenetic analysis confirmed that isolate KY11 clusters within the same clade as F. incarnatum, thereby supporting its taxonomic classification. (Garcia-Estrada et al.,2021). Isolate KY11 was identified as F. incarnatum based on morphological and sequence analyses. Pathogenicity tests were conducted on 3-month-old walnut seedlings to fulfill Koch's postulates. The inoculation procedure was as follows: three fully expanded healthy leaves per seedling were surface-sterilized with 75% ethanol, wounded with a sterile needle, and inoculated with a KY11 spore suspension at a concentration of 106 spores/mL. Control plants were mock-inoculated with sterile water. All plants were maintained at 28°C with 70% relative humidity under a 12-h light/dark cycle. After 12 days, inoculated leaves developed symptoms identical to field observations, while controls remained asymptomatic. The experiment consisted of two independent trials with five seedlings each. F. incarnatum was successfully re-isolated from symptomatic leaves and confirmed through both morphological and molecular methods, thereby fulfilling Koch's postulates and verifying its pathogenicity. This pathogen has been reported to cause cucumber fruit rot in Mexico (Garcia-Estrada et al., 2021), maize leaf blight (Xu et al., 2022), and luffa leaf spot (Chen et al., 2025) in China. To our knowledge, this represents the first report of F. incarnatum causing walnut brown spot in China, providing fundamental information for disease diagnosis and the development of control strategies.