Four new species of Dothideomycetes (Ascomycota) from Pará Rubber (Heveabrasiliensis) in Yunnan Province, China.

The tropical areas in southern and south-western Yunnan are rich in fungal diversity. Additionally, the diversity of seed flora in Yunnan Province is higher than in other regions in China and the abundant endemic species of woody plants provide favourable substrates for fungi. Rubber plantations in Yunnan Province are distributed over a large area, especially in Xishuangbanna. During a survey of rubber-associated fungi in Yunnan Province, China, dead rubber branches with fungal fruiting bodies were collected. Morphological characteristics and multigene phylogenetic analyses (ITS, LSU, SSU, rpb2 and tef1-α) revealed four distinct new species, described herein as Melomastiapuerensis, Nigrogranalincangensis, Pseudochaetosphaeronemalincangensis and Pseudochaetosphaeronemaxishuangbannaensis. Detailed descriptions, illustrations and phylogenetic trees are provided to show the taxonomic placements of these new species.

Comparative genomics reveals insight into the phylogeny and habitat adaptation of novel Amycolatopsis species, an endophytic actinomycete associated with scab lesions on potato tubers.

A novel endophytic actinomycete, strain MEP2-6T, was isolated from scab tissues of potato tubers collected from Mae Fag Mai Sub-district, San Sai District, Chiang Mai Province, Thailand. Strain MEP2-6T is a gram-positive filamentous bacteria characterized by meso-diaminopimelic acid in cell wall peptidoglycan and arabinose, galactose, glucose, and ribose in whole-cell hydrolysates. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and hydroxy-phosphatidylethanolamine were the major phospholipids, of which MK-9(H6) was the predominant menaquinone, whereas iso-C16:0 and iso-C15:0 were the major cellular fatty acids. The genome of the strain was 10,277,369 bp in size with a G + C content of 71.7%. The 16S rRNA gene phylogenetic and core phylogenomic analyses revealed that strain MEP2-6T was closely related to Amycolatopsis lexingtonensis NRRL B-24131T (99.4%), A. pretoriensis DSM 44654T (99.3%), and A. eburnea GLM-1T (98.9%). Notably, strain MEP2-6T displayed 91.7%, 91.8%, and 87% ANIb and 49%, 48.8%, and 35.4% dDDH to A. lexingtonensis DSM 44653T (=NRRL B-24131T), A. eburnea GLM-1T, and A. pretoriensis DSM 44654T, respectively. Based on phenotypic, chemotaxonomic, and genomic data, strain MEP2-6T could be officially assigned to a novel species within the genus Amycolatopsis, for which the name Amycolatopsis solani sp. nov. has been proposed. The type of strain is MEP2-6T (=JCM 36309T = TBRC 17632T = NBRC 116395T). Amycolatopsis solani MEP2-6T was strongly proven to be a non-phytopathogen of potato scab disease because stunting of seedlings and necrotic lesions on potato tuber slices were not observed, and there were no core biosynthetic genes associated with the BGCs of phytotoxin-inducing scab lesions. Furthermore, comparative genomics can provide a better understanding of the genetic mechanisms that enable A. solani MEP2-6T to adapt to the plant endosphere. Importantly, the strain smBGCs accommodated 33 smBGCs encoded for several bioactive compounds, which could be beneficially applied in the fields of agriculture and medicine. Consequently, strain MEP2-6T is a promising candidate as a novel biocontrol agent and antibiotic producer.

Distinct fungal microbiomes of two Thai commercial stingless bee species, Lepidotrigona terminata and Tetragonula pagdeni suggest a possible niche separation in a shared habitat.

Stingless bees, a social corbiculate bee member, play a crucial role in providing pollination services. Despite their importance, the structure of their microbiome, particularly the fungal communities, remains poorly understood. This study presents an initial characterization of the fungal community associated with two Thai commercial stingless bee species, Lepidotrigona terminata (Smith) and Tetragonula pagdeni (Schwarz) from Chiang Mai, Thailand. Utilizing ITS amplicon sequencing, we identified distinct fungal microbiomes in these two species. Notably, fungi from the phyla Ascomycota, Basidiomycota, Mucoromycota, Mortierellomycota, and Rozellomycota were present. The most dominant genera, which varied significantly between species, included Candida and Starmerella. Additionally, several key enzymes associated with energy metabolism, structural strength, and host defense reactions, such as adenosine triphosphatase, alcohol dehydrogenase, ß-glucosidase, chitinase, and peptidylprolyl isomerase, were predicted. Our findings not only augment the limited knowledge of the fungal microbiome in Thai commercial stingless bees but also provide insights for their sustainable management through understanding their microbiome.

Improving furfural tolerance in a xylose-fermenting yeast Spathaspora passalidarum CMUWF1-2 via adaptive laboratory evolution.

BACKGROUND: Spathaspora passalidarum is a yeast with the highly effective capability of fermenting several monosaccharides in lignocellulosic hydrolysates, especially xylose. However, this yeast was shown to be sensitive to furfural released during pretreatment and hydrolysis processes of lignocellulose biomass. We aimed to improve furfural tolerance in a previously isolated S. passalidarum CMUWF1-2, which presented thermotolerance and no detectable glucose repression, via adaptive laboratory evolution (ALE). RESULTS: An adapted strain, AF2.5, was obtained from 17 sequential transfers of CMUWF1-2 in YPD broth with gradually increasing furfural concentration. Strain AF2.5 could tolerate higher concentrations of furfural, ethanol and 5-hydroxymethyl furfuraldehyde (HMF) compared with CMUWF1-2 while maintaining the ability to utilize glucose and other sugars simultaneously. Notably, the lag phase of AF2.5 was 2 times shorter than that of CMUWF1-2 in the presence of 2.0 g/l furfural, which allowed the highest ethanol titers to be reached in a shorter period. To investigate more in-depth effects of furfural, intracellular reactive oxygen species (ROS) accumulation was observed and, in the presence of 2.0 g/l furfural, AF2.5 exhibited 3.41 times less ROS accumulation than CMUWF1-2 consistent with the result from nuclear chromatins diffusion, which the cells number of AF2.5 with diffuse chromatins was also 1.41 and 1.24 times less than CMUWF1-2 at 24 and 36 h, respectively. CONCLUSIONS: An enhanced furfural tolerant strain of S. passalidarum was achieved via ALE techniques, which shows faster and higher ethanol productivity than that of the wild type. Not only furfural tolerance but also ethanol and HMF tolerances were improved.

Sustainable Innovation: Fabrication and Characterization of Mycelium-Based Green Composites for Modern Interior Materials Using Agro-Industrial Wastes and Different Species of Fungi.

Mycelium-based bio-composites (MBCs) represent a sustainable and innovative material with high potential for contemporary applications, particularly in the field of modern interior design. This research investigates the fabrication of MBCs for modern interior materials using agro-industrial wastes (bamboo sawdust and corn pericarp) and different fungal species. The study focuses on determining physical properties, including moisture content, shrinkage, density, water absorption, volumetric swelling, thermal degradation, and mechanical properties (bending, compression, impact, and tensile strength). The results indicate variations in moisture content and shrinkage based on fungal species and substrate types, with bamboo sawdust exhibiting lower shrinkage. The obtained density values range from 212.31 to 282.09 kg/m3, comparable to traditional materials, suggesting MBCs potential in diverse fields, especially as modern interior elements. Water absorption and volumetric swelling demonstrate the influence of substrate and fungal species, although they do not significantly impact the characteristics of interior decoration materials. Thermal degradation analysis aligns with established patterns, showcasing the suitability of MBCs for various applications. Scanning electron microscope observations reveal the morphological features of MBCs, emphasizing the role of fungal mycelia in binding substrate particles. Mechanical properties exhibit variations in bending, compression, impact, and tensile strength, with MBCs demonstrating compatibility with traditional materials used in interior elements. Those produced from L. sajor-caju and G. fornicatum show especially promising characteristics in this context. Particularly noteworthy are their superior compression and impact strength, surpassing values observed in certain synthetic foams multiple times. Moreover, this study reveals the biodegradability of MBCs, reaching standards for environmentally friendly materials. A comprehensive comparison with traditional materials further supports the potential of MBCs in sustainable material. Challenges in standardization, production scalability, and market adoption are identified, emphasizing the need for ongoing research, material engineering advancements, and biotechnological innovations. These efforts aim to enhance MBC properties, promoting sustainability in modern interior applications, while also facilitating their expansion into mass production within the innovative construction materials market.

Bacterial diversity, community structure and function in association of potato scabby tubers during storage in northern Thailand.

Potato scab is a common potato tuber disease that affects quality and cost in the marketplace, shortening storage, and increasing the chance for secondary infection. The tubers with disease severity of 1 to 4 are accepted and stored in potato storage for cheap selling in Thailand. However, there are few studies of the bacterial community of the scabby tuber during storage. Thus, we aim to elucidate the diversity, structure, and function of the bacterial community of 30-day storage potato scabby tubers stored in different temperatures using 16S amplicon metagenomic sequencing. Bacterial communities of storage potato scabby tubers (Spunta cultivar) collected from different storage temperatures, 4 °C (MEP1) and 6 °C (MEP2), were characterized using 16S rRNA amplicon metagenomic sequencing. The alpha-diversity abundance in the bacteriome of the scabby tubers stored at 6 °C was higher than in those stored at 4 °C. Actinobacteria (34.7%) was a dominant phylum in MEP1, while Proteobacteria (39.9%) was predominant in MEP2. The top 10 genera of both communities were Rhizobium group, Streptomyces, Pectobacterium, Ruminococcus, Cellulomonas, Promicromonospora, Prevotella, Enterobacter, Pedobacter, and Paenarthrobacter. Moreover, functional profile prediction of both communities reveals essential genes in the pathosystem: nos, bglA, and cebEFG-msiK for potato scab disease and phc and peh operons for rot disease. Our findings are the first study to explore details of the bacteriome of the accepted potato scabby tubers for selling during storage in Thailand and strongly indicate that although potatoes were stored at low temperatures, diseases still occur by secondary pathogens.

A New Edible Wild Mushroom Species, Panus sribuabanensis (Panaceae, Polyporales) from Northern Thailand and Its Nutritional Composition, Total Phenolic Content, and Antioxidant Activity.

A new edible wild mushroom species, described herein as Panus sribuabanensis, was collected from local markets and natural forests located in northern Thailand. This species is characterized by its medium to large-sized basidiomata, broadly ellipsoid to ellipsoid-shaped basidiospores, dimitic hyphal system, and the absence of hyphal pegs. A molecular phylogenetic analysis of combined the internal transcribed spacer (ITS) and large subunit (nrLSU) of nuclear ribosomal DNA sequences supported the monophyly of P. sribuabanensis as a distinct lineage within the genus Panus. Full description, illustrations, color photographs, and a phylogenetic tree to show the placement of P. sribuabanensis are provided. The dried mushroom showed a nutritional composition within the range of 2.58%-2.67% for fat content, 27.10%-27.98% for protein, and 43.97%-44.10% for carbohydrates. The ethanolic extracts from this mushroom exhibited a total phenolic content ranging from 0.66 to 0.74 mg GAE/g dry weight (dw). Moreover, the antioxidant activities of ethanolic extracts evaluated by the 2,2-diphenyl-1-picrylhydrazyl (0.90-1.08 mg TE/g dw) and ferric reducing antioxidant power (0.93-1.08 mg TE/g dw) assays demonstrate higher activity compared to the 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay (0.44-0.51 mg TE/g dw). The outcomes of this study provide significant information on the nutritional value, phenolic content, and antioxidant activity potential of this new mushroom species discovered in northern Thailand.

Additions to Diatrypaceae (Xylariales): Novel Taxa and New Host Associations.

Diatrypaceae members have a broad distribution and are commonly found on decaying wood. Despite taxonomic and morphological challenges within this group, there has been a growing interest in Diatrypaceae in recent years. The dead branches of several plant hosts with fungal fruiting bodies were collected from Doi Tung National Park, Chiang Rai, and the Narathiwat Provinces in Thailand. Their morphological characteristics, coupled with a molecular phylogeny of combined ITS and tub2 sequence data, were used to introduce two novel Allodiatrype species (A. dalbergiae and A. eleiodoxae) and one new Melanostictus species (M. chiangraiensis). Moreover, four new host records, Diatrypella heveae, D. major, Melanostictus thailandicus, and Paraeutypella citricola on Microcos paniculata, Nayariophyton zizyphifolium, Dalbergia cultrata, and M. paniculata, respectively, as well as a new geographical record of D. major are reported. This research provides detailed descriptions of macro- and microcharacteristics, coupled with a phylogenetic tree for the newly introduced species and host records. The morphological features of Allodiatrype and Melanostictus are listed in the synoptic table.

Addition of Five Novel Fungal Flora to the Xylariomycetidae (Sordariomycetes, Ascomycota) in Northern Thailand.

The deviation of conventional fungal niches is an important factor in the implications of hidden fungal diversity and global fungal numbers. The Xylariomycetidae (Sordariomycetes, Ascomycota), which is also referred to as xylarialean taxa, has a wide range of species that demonstrate a high degree of variation in their stromatic characteristics, showing either conspicuous or inconspicuous forms. In this study, samples were collected while focusing on temporal and spatial parameters and substrate characteristics. Based on internal transcribed spacer (ITS), 28S large subunit rDNA (LSU), RNA polymerase II second largest subunit (RPB2), and ß-tubulin (TUB2) multigene phylogeny and morphology, five new species are introduced as Muscodor brunneascosporus, M. lamphunensis (Xylariaceae), Nigropunctata hydei, N. saccata (Incertae sedis), and Xenoanthostomella parvispora (Gyrotrichaceae). Plant substrates in the early stages of decay and attached to the host were feasible sample niches, with an emphasis on the collection of inconspicuous, hidden xylarialean species. The appearance of inconspicuous saprobic xylarialean forms during the rainy season may be linked to the change in nutritional mode, from endophytic mode during the dry season to saprobic in the wet. Therefore, it would be fascinating to concentrate future research on how seasonal fluctuations affect nutritional mode shifts, especially in northern Thailand, which would provide the optimal spatial characteristics. In order to establish a comprehensive linkage between endophytic and saprobic modes, it is imperative to have a substantial representation of endophytic isolate sequences resembling inconspicuous xylariaceous fungi within publicly accessible databases.

Multifarious Characterization and Efficacy of Three Phosphate-Solubilizing Aspergillus Species as Biostimulants in Improving Root Induction of Cassava and Sugarcane Stem Cuttings.

Several soil fungi significantly contribute to the enhancement of plant development by improving nutrient uptake and producing growth-promoting metabolites. In the present study, three strains of phosphate-solubilizing fungi, namely, Aspergillus chiangmaiensis SDBR-CMUI4, A. pseudopiperis SDBR-CMUI1, and A. pseudotubingensis SDBR-CMUO2, were examined for their plant-growth-promoting capabilities. The findings demonstrated that all fungi showed positive siderophore production, but only A. pseudopiperis can produce indole-3-acetic acid. All fungi were able to solubilize insoluble phosphate minerals [Ca3(PO4)2 and FePO4] by producing phosphatase enzymes and organic acids (oxalic, tartaric, and succinic acids). These three fungal species were grown at a water activity ranging from 0.837 to 0.998, pH values ranging from 4 to 9, temperatures between 4 and 40 °C, and 16-17% NaCl in order to evaluate their drought, pH, temperature, and salt tolerances, respectively. Moreover, the results indicated that A. pseudopiperis and A. pseudotubingensis were able to tolerate commercial insecticides (methomyl and propargite) at the recommended dosages for field application. The viability of each fungal strain in the inoculum was higher than 50% at 4 and 20 °C after 3 months of storage. Subsequently, all fungi were characterized as plant-growth-promoting strains by improving the root inductions of cassava (Manihot esculenta Crantz) and sugarcane (Saccharum officinarum L.) stem cuttings in greenhouse experiments. No symptoms of plant disease were observed with any of the treatments involving fungal inoculation and control. The cassava and sugarcane stem cuttings inoculated with fungal strains and supplemented with Ca3(PO4)2 exhibited significantly increased root lengths, shoot and root dry biomasses, chlorophyll concentrations, and cellular inorganic phosphate contents. Therefore, the application of these phosphate-solubilizing fungi is regarded as a new frontier in the induction of roots and the promotion of growth in plants.

Life on the wall: the diversity and activity of microbes on 13th - century AD. Lan Na mural painting.

Diverse microorganisms from the three domains of life (Archaea, Bacteria, and Eukaryota) cause deterioration in mural paintings worldwide; however, few studies have simultaneously targeted these three domains. This study aims to survey the microbiome and its potential for biodeterioration on unpreserved Lan Na mural paintings in Sean Khan temple, Chiang Mai, Thailand. The overview of the archaeal, bacterial, and fungal communities was reported by Illumina sequencing, whereas the potential for deterioration was revealed by culturable techniques and a literature search. The abundant microbes reported in this study were also found in other ancient mural paintings worldwide. Halococcus, a salt-tolerant archaeon, as well as the eubacterial genus Crossiella dominated the prokaryotic community. On the other hand, the main fungal group was the genus Candida (Ascomycota). However, a low number of fungi and bacteria were isolated. Most of the isolates showed the ability to survive in the drought conditions of mural paintings but could not perform discoloration activities. The deterioration activity mainly affected calcium compounds, which are the main components of painting substrates. Aspergillus and several bacterial isolates could dissolve calcium compounds, but only Trichaptum species could induce crystal formation. These results suggest that deterioration of painting substrate should be taken into consideration in addition to deterioration of color in mural paintings. For the Lan Na painting in Sean Khan temple, the plaster is the prime target for biodeterioration, and thus we suggest that the preservation effort should focus on this component of the mural painting.

Value-added green biorefinery co-products from ultrasonically assisted DES-pretreated Chlorella biomass.

This study pursued the goal of creating value-added co-products through an environmentally friendly biorefinery approach, employing ultrasonically assisted deep eutectic solvent (DES)-pretreated Chlorella biomass. The primary focus was on generating enriched biodiesel feedstock with exceptional fuel properties and developing hydroponic biofertilizer. The results demonstrated the effectiveness of a two-step process involving a 5-minute ultrasound-assisted DES pretreatment followed by ultrasound-assisted solvent extraction, which efficiently extracted lipids from Chlorella biomass, yielding biodiesel-quality lipids with good cetane number (59.42) and high heating value (40.11 MJ/kg). Notably, this two-step approach (78.04 mg-lipid/g-microalgal biomass) led to a significant 2.10-fold increase in lipid extraction compared to a one-step process (37.15 mg-lipid/g-microalgal biomass) that combined ultrasound-assisted DES pretreatment and solvent extraction. Importantly, the aqueous extract derived from lipid-extracted microalgal biomass residues (LMBRs) showed promise as a component in hydroponic biofertilizer production, supporting lettuce growth in hydroponic deep water culture system. Consequently, microalgae biorefinery co-products hold tremendous potential in enhancing the profitability and sustainability of interconnected sectors, encompassing renewable energy, agriculture, and the environment.

Exploring diversity rock-inhabiting fungi from northern Thailand: a new genus and three new species belonged to the family Herpotrichiellaceae.

Members of the family Herpotrichiellaceae are distributed worldwide and can be found in various habitats including on insects, plants, rocks, and in the soil. They are also known to be opportunistic human pathogens. In this study, 12 strains of rock-inhabiting fungi that belong to Herpotrichiellaceae were isolated from rock samples collected from forests located in Lamphun and Sukhothai provinces of northern Thailand during the period from 2021 to 2022. On the basis of the morphological characteristics, growth temperature, and multi-gene phylogenetic analyses of a combination of the internal transcribed spacer, the large subunit, and the small subunit of ribosomal RNA, beta tubulin and the translation elongation factor 1-a genes, the new genus, Petriomyces gen. nov., has been established to accommodate the single species, Pe. obovoidisporus sp. nov. In addition, three new species of Cladophialophora have also been introduced, namely, Cl. rupestricola, Cl. sribuabanensis, and Cl. thailandensis. Descriptions, illustrations, and a phylogenetic trees indicating the placement of these new taxa are provided. Here, we provide updates and discussions on the phylogenetic placement of other fungal genera within Herpotrichiellaceae.

Exploring ascomycete diversity in Yunnan, China I: resolving ambiguous taxa in Phaeothecoidiellaceae and investigating conservation implications of fungi.

Yunnan, located in southwestern China, is known for its high fungal diversity, and many of which are endemic to the region. As part of our ongoing studies on fungi in Yunnan, we introduce two new genera in Phaeothecoidiellaceae (Mycosphaerellales), to accommodate one Repetophragma-like and another Stomiopeltis-like taxa. Pseudorepetophragma gen. nov. is introduced herein as a monotypic genus to accommodate P. zygopetali comb. nov.(≡ Repetophragma zygopetali), whereas Pseudostomiopeltis gen. nov. is introduced to accommodate Ps. xishuangbannaensis gen. et sp. nov. and Ps. phyllanthi comb. nov.(≡ Stomiopeltis phyllanthi), based on a new collection from Yunnan. In addition, Stomiopeltis sinensis is transferred to Exopassalora as E. sinensis comb. nov. due to its phylogenetic affinity and grouped with E. zambiae, the generic type of Exopassalora. This study provides new insights into the biodiversity of fungal species in this region and adds to our understanding of their ecological roles, as well as the resolution to ambiguous taxa in Phaeothecoidiellaceae.

The Potential of Endophytic Fungi for Enhancing the Growth and Accumulation of Phenolic Compounds and Anthocyanin in Maled Phai Rice (Oryza sativa L.).

Rice (Oryza sativa L.) is one of the most popular cereal crops, being consumed by almost half of the world's population. Among several cultivars widely distributed in Thailand, Maled Phai is a Thai pigmented-upland rice with exceptionally high nutritional value and high demand in the local Thai market. This study aimed to investigate the feasibility of producing plant growth-promoting properties (PGP) and enhancing the accumulation of phytochemicals in Maled Phai rice seeds of endophytic fungi isolated from upland black rice. Among a total of 56 isolates, the 4 most effective PGP isolates were identified as Trichoderma zelobreve PBMP16, Talaromyces pinophilus PBMP28, Aspergillus flavus KKMP34, and Trichoderma sp. PBMP4 based on their morphological characteristics and multigene (ITS, rpb2, tef-1, CaM, and BenA) phylogenetic analyses. These four endophytic fungi could promote plant growth parameters under greenhouse conditions. Outstandingly, upland rice inoculated with Tr. zelobreve PBMP16 had a significant increase in total seed weight, root length, phenolic compounds, anthocyanin, antioxidants, and N uptake, which were higher than those of the noninoculated control, and even better than the chemical fertilizer. Overall, this report shows that endophytic fungi efficiently promoted growth and increased the phenolic compounds, anthocyanin, and antioxidants of Maled Phai rice.

Seasonal Variations in Fungal Communities on the Surfaces of Lan Na Sandstone Sculptures and Their Biodeterioration Capacities.

Environmental factors and climate are the primary factors influencing the microbial colonization and deterioration of cultural heritage in outdoor environments. Hence, it is imperative to investigate seasonal variations in microbial communities and the biodeterioration they cause. This study investigated the surfaces of sandstone sculptures at Wat Umong Suan Phutthatham, Chiang Mai, Thailand, during wet and dry seasons using culture-dependent and culture-independent approaches. The fungi isolated from the sandstone sculptures were assessed for biodeterioration attributes including drought tolerance, acid production, calcium crystal formation, and calcium precipitation. The results show that most of the fungal isolates exhibited significant potential for biodeterioration activities. Furthermore, a culture-independent approach was employed to investigate the fungal communities and assess their diversity, interrelationship, and predicted function. The fungal diversity and the communities varied seasonally. The functional prediction indicated that pathotroph-saprotroph fungi comprised the main fungal guild in the dry season, and pathotroph-saprotroph-symbiotroph fungi comprised the dominant guild in the wet season. Remarkably, a network analysis revealed numerous positive correlations among fungal taxa within each season, suggesting a potential synergy that promotes the biodeterioration of sandstone. These findings offer valuable insights into seasonal variations in fungal communities and their impacts on the biodeterioration of sandstone sculptures. This information can be utilized for monitoring, management, and maintenance strategies aimed at preserving this valuable cultural heritage.

First Report of Anthracnose on Giant Philodendron Caused by Colletotrichum siamense in Thailand.

Giant philodendron (Philodendron giganteum Schott) is cultivated in Thailand and has become an important ornamental houseplant with great economic value. During the rainy season in July 2022, anthracnose disease on this plant was observed at a nursery in Saraphi District, Chiang Mai Province (18°40'18" N, 99°03'17" E), Thailand. The area investigated was approximately 800 m². The disease incidence was estimated at above 15% according to the total number of plants (220 plants). The disease severity of each plant was between 25 and 50% of the necrotic lesion on the leaf. Initially, symptoms with brown spots, appeared on leaves, gradually becoming enlarged, elongate, 1 to 11 cm long by 0.3 to 3.5 cm wide, irregular, sunken, dark brown, with a yellow halo surrounding each lesion. Then, the diseased leaves eventually withered and died. Leaf pieces (5 × 5 mm2) of the margins between lesions and the healthy tissue were surface sterilized in 1% NaClO for 1 min, 70% ethanol for 30 s, and rinsed three times with sterile distilled water. Tissues were placed on potato dextrose agar (PDA) and incubated at 25°C in darkness. After three days of incubation, pure fungal colonies were purified by a single hyphal tip method on PDA (Korhonen and Hintikka 1980). Two fungal isolates (SDBR-CMU471 and SDBR-CMU472) with similar morphology were obtained. Fungal colonies on PDA were white and 38 to 40 mm in diameter after 3 days of incubation at 25 °C, then grayish white with cottony mycelia, the reverse side pale yellow after one week of incubation. Both isolates produced asexual structures on PDA. Setae were brown with 1 to 3 septa, 50 to 110 × 2.4 to 4.0 µm, with a cylindrical base, and acuminate tip. Conidiophores were hyaline to pale brown, septate, and branched. Conidiogenous cells were hyaline to pale brown, cylindrical to ampulliform, 9.5 to 35 µm long (n = 50). Conidia were single-celled, straight, hyaline, smooth-walled, cylindrical, ends rounded, guttulate, 9.1 to 19.6 × 3.5 to 5.6 µm (n = 50). Appressoria were brown to dark brown, oval to irregular, smooth-walled, 5 to 10 × 5 to 7.5 µm (n = 50). Morphologically, both fungal isolates resembled members of the Colletotrichum gloeosporioides species complex (Weir et al. 2012; Jayawardena et al. 2021). The internal transcribed spacer (ITS) region of the ribosomal DNA, actin (act), ß-tubulin (tub2), calmodulin (CAL), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified using primer pairs ITS5/ITS4 (White et al. 1990), ACT-512F/ACT-783R (Carbone and Kohn 1999), T1/T22 (O'Donnell and Cigelnik 1997), CL1C/CL2C (Weir et al. 2012), and GDF1/GDR1 (Templeton et al. 1992), respectively. Sequences were deposited in GenBank (ITS: OQ699280, OQ699281; act: OQ727122, OQ727123; tub2: OQ727124, OQ727125; CAL: OQ727126, OQ727127; GAPDH: OQ727128, OQ727129). Multi-gene (combined data set of ITS, GAPDH, CAL, act, and tub2) maximum likelihood phylogenetic analyses demonstrated that both isolates were identified as C. siamense with 100% support. In a pathogenicity test, leaves of healthy plants were surface sterilized with a 0.1% NaClO solution for 3 min, rinsed three times with sterile distilled water. After being air-dried, a uniform wound (5 pores, 3 mm in width) was made at the equator of each leaf using aseptic needles. Conidial suspensions were collected from two-week-old cultures and suspended in sterile distilled water with 0.05% Tween-20. Fifteen microliters of the conidial suspension (1 × 106 conidia/ml) were placed on wounded attached leaves. As well, wounded control leaves were mock inoculated with sterile distilled water. Ten replications were conducted for each treatment and experiments were repeated twice. The inoculated plants were stored in a greenhouse at conditions of 25 to 30°C and 75 to 85% relative humidity. After 14 days, all the inoculated leaves showed disease symptoms, brown lesions with yellow halos, whereas control leaves remained asymptomatic. The pathogen C. siamense was consistently re-isolated on PDA from the inoculated tissues to complete Koch's postulates. Colletotrichum siamense has been reported as a causal agent on a wide range of host plants in Thailand and throughout the world (Farr and Rossman 2021; Jayawardena et al. 2021). Prior to this study, C. endophytica, C. karsti, C. orchidearum, C. philodendricola, and C. pseudoboninense were identified as causal agents of anthracnose on philodendrons (Xue et al. 2020; Zhang et al. 2023). However, anthracnose caused by Colletotrichum species on giant philodendron (P. giganteum) has not been previously reported. Thus, we propose C. siamense as a new causal agent of anthracnose disease on giant philodendron. This study provides information for further investigation into the epidemiology and management of this disease. Moreover, further investigations should be carried out in other philodendron growing areas of Thailand in order to specifically search for this pathogen.

Taxonomic Novelties and New Records of Amanita Subgenus Amanitina from Thailand.

The Amanita subgenus Amanitina contains six sections, and the species diversity of this subgenus has still not been explored in Thailand. Twenty samples collected in 2019 and 2020, which had the morphological characteristics of the Amanita subgen. Amanitina, were observed in this study. Both the microscopical characteristics and multi-gene phylogenetic analyses of the ITS, nrLSU, RPB2, TEF1-α, and TUB gene regions revealed that the 20 samples represented nine species and dispersed into four sections. Remarkably, three taxa were different from any other currently known species. Here, we describe them as new to science, namely A. albifragilis, A. claristriata, and A. fulvisquamea. Moreover, we also recognized six interesting taxa, including four records that were new to Thailand, viz. A. cacaina, A. citrinoannulata, A. griseofarinosa, and A. neoovoidea, as well as two previously recorded species, A. caojizong and A. oberwinkleriana. Moreover, we provide the first RPB2 and TEF1-α gene sequences for A. cacaina. Detailed descriptions, illustrations as line drawings, and comparisons with related taxa are provided.

Insights into Diversity, Distribution, and Systematics of Rust Genus Puccinia.

Puccinia, which comprises 4000 species, is the largest genus of rust fungi and one of the destructive plant pathogenic rust genera that are reported to infect both agricultural and nonagricultural plants with severe illnesses. The presence of bi-celled teliospores is one of the major features of these rust fungi that differentiated them from Uromyces, which is another largest genus of rust fungi. In the present study, an overview of the current knowledge on the general taxonomy and ecology of the rust genus Puccinia is presented. The status of the molecular identification of this genus along with updated species numbers and their current statuses in the 21st century are also presented, in addition to their threats to both agricultural and nonagricultural plants. Furthermore, a phylogenetic analysis based on ITS and LSU DNA sequence data available in GenBank and the published literature was performed to examine the intergeneric relationships of Puccinia. The obtained results revealed the worldwide distribution of Puccinia. Compared with other nations, a reasonable increase in research publications over the current century was demonstrated in Asian countries. The plant families Asteraceae and Poaceae were observed as the most infected in the 21st century. The phylogenetic studies of the LSU and ITS sequence data revealed the polyphyletic nature of Puccinia. In addition, the presences of too short, too lengthy, and incomplete sequences in the NCBI database demonstrate the need for extensive DNA-based analyses for a better understanding of the taxonomic placement of Puccinia.