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For this molecular genetic research, helminthological samples were collected from the lungs of 17 samples of Pelophylax terentievi (Mezhzherin, 1992) distributed in Kashkadarya and Surkhondarya regions in the spring and summer of 2024. The difference between the nucleotides of the R. engelbrechti_uz (PQ219671) sample belonging to the Rhabdias genus collected from the southern regions of our republic and the R. engelbrechti (MG428406) sample obtained from the NCBI database was 0.4%. Nucleotide sequence obtained as a result of molecular-genetic study of engelbrechti belonging to Rhabdias genus was placed in the National Center for Biotechnology Information. This species was recorded for the first time to the best of our knowledge in the territory of the Republic of Uzbekistan. [ABSTRACT FROM AUTHOR]

Nowadays, diseases are a major concern in fish production, particularly in gills, which play a crucial role in blood oxygenation, acid-base balance, osmoregulation, and waste elimination. Fungal infections, particularly Branchiomyces, cause respiratory problems in fish. This study aims to determine the causative agent causing high mortality in common carp, Cyprinus carpio, in the middle Euphrates in Iraq. One hundred infected fish were sampled for microbial, histological, and molecular examination. Gill tissue was examined as well for histological alterations, and DNA was isolated from Branchiomyces and amplified with universal primers. Microscopically identifying Branchiomyces spp. using lactophenol staining, non-separated hyphae and spores (5–9 m in size) at various stages inside the structures of gill tissue were present. Histopathological examination revealed hyperplasia of epithelial cells and infusion of gill filaments, while primary gill filaments displayed severe hemorrhage and edema. Hyphae of Branchiomyces have been detected between necrotic and edematous myofiber. The ITS PCR products of the fungal isolate were found to be positive at 540 bp molecular weight. Economic losses can occur as a result of fish infections caused by Branchiomyces. Histopathological lesions in gill tissue indicated the presence of Branchiomyces infection, which was confirmed by molecular and microscopical examination.

The American cranberry is a perennial North American fruit plant that is grown successfully on commercial plantations in Poland. The purpose of this study was to recognize filamentous fungi that colonize roots, leaves, and fruits without visible disease symptoms. Pure fungal cultures were isolated from disinfected plant fragments in agar media and identified by sequencing common taxonomic DNA markers such as the ITS region, the TEF-1α, or RPB2 genes. Of the 141 isolates studied, 59% were identified as closely related to soil saprotrophs. They were classified primarily as showing the greatest similarity to type strains of Trichoderma amoenum, Trichoderma dorothopsis, Paraphaeosphaeria sporulosa, and Penicillium murcianum. Additionally, isolates that are most similar to strains of Penicillium crustosum, Aspergillus flavus, and Aspergillus versicolor that produced mycotoxins were detected. The fungi identified as closest to Alternaria geophila, Alternaria senecionicola, Paraphoma radicina, Pestalotiopsis unicolor, Pestalotiopsis scoparia, and Neopestalotiopsis spp., whose hosts are plants other than American cranberry, represented 33.81% of the isolates tested. Only 7.2% of the isolates corresponded to the species of Physalospora vaccinia, Diaporthe vaccinii, and Diaporthe eres, known cranberry pathogens. The results of this study can be used to identify latent plant infection and potential disease risks. [ABSTRACT FROM AUTHOR]

Truffles represent a resource of great value all over the world. The various species are widespread and occupy various habitats. Methods to identify their value in terms of marketing, cultivation, and the ecological–forestal field are needed. The traditional classification of Tuber species is based on their morphology; nevertheless, the macroscopic and microscopic characteristics of the fruiting body, spores, and ectomycorrhizae may be absent or insufficiently discriminating. This problem is amplified when dealing with valuable truffle species, which are often replaced by truffles with lower organoleptic qualities and commercial value. Today, molecular biology can help to correctly identify them through several widely used methods that are fast, specific, and sensitive. Molecular methods are mostly PCR-based, with primers designed on various targets, genes, or regions widely variable between species but sufficiently conserved within the same species. The DNA regions analyzed for identifying truffles include ribosomal internal transcribed spacer (ITS), ribosomal large subunit (LSU), β-tubulin, and EF-α elongation factor. This review summarizes the main and most reliable DNA-based molecular methods for identifying Tuber species. The methods are listed schematically and clearly for quick consultation and use in all areas of study in which monitoring, and verification of the species is essential.

Myxobolus nagaraensis is a myxozoan parasite first reported in freshwater gobies (Rhinogobius spp.) from the Nagara River, Gifu Prefecture, Japan. Myxospores of M. nagaraensis form plasmodia in the visceral cavities of gobies, commonly presenting as distended abdomens. Although Rhinogobius is a common fish genus in Japan, details of M. nagaraensis, including genetic information, remain unknown. We compared the nucleotide sequences of the ribosomal RNA gene (rDNA) of M. nagaraensis from three different host species (R. fluviatilis, R. nagoyae, and R. similis) caught in three different rivers in Japan (Sakai, Sagami, and Kaname). The ITS region (ITS-1, 5.8S rDNA, and ITS-2) and large subunit (LSU) rDNA exhibited 49 and 55 variable sites, respectively. The highest nucleotide diversity was observed in the ITS region (0.00962), whereas that of the LSU rDNA was 0.00187. Differences in host species, rather than rivers, were a significant factor for genetic variation in both the ITS region (62.58%; P

Endophytic fungi colonize the inner tissues and provide direct and indirect benefits to plants. Although Mongolia is rich in medicinal plants, due to climatic and anthropogenic reasons, the resources are being depleted, and many species are under threat of gradual extinction, while the endophytic fungi of Mongolian plants are largely unknown. In this study, a total of 24 culturable endophytic fungal strains were isolated from Cynomorium songaricum (Rupr.), a medicinal and vulnerable plant species of Mongolia. Based on the morphological characteristics and the sequences of the rDNA internal transcribed spacer (ITS) region, the isolates were identified into six genera: Fusarium (8), Clonostachys (7), Penicillium (6), Alternaria (1), Aspergillus (1), and Madurella (1). The antimicrobial activity was assessed by the agar-diffusion method, revealing that 15 strains were able to inhibit the growth of at least one of the test organisms. Among them, 1 strain showed inhibitory activity against Escherichia coli, 12 against Bacillus subtilis, 13 against Staphylococcus aureus, and 8 against Aspergillus niger, respectively. The ability to solubilize complex phosphorus and zinc minerals was observed in 3 and 21 strains, respectively, and the production of indole-3-acetic acid (IAA) was detected in nine strains in the presence of tryptophan. Our study provides the first insight into the cultivable endophytic fungal composition of C. songaricum, parasitizing the roots of Nitraria sibirica growing in the Gobi Desert of Mongolia. The resulting fungi, which have antimicrobial and plant growth-promoting properties, were preserved in the national culture collection and can be used to further exploit their biotechnological potential, as well as for the propagation of endangered and vulnerable medicinal plants. [ABSTRACT FROM AUTHOR]

Since Mulberries (Morus)is a tree species with a considerable plant variety. Molecular techniques are methods used to distinguish between species accurately, easily and quickly. This study examines a Molecular method for distinguishing different Morus species in the Duhok - Kurdistan region/ Iraq. The method is based on the use of four techniques: matK gene, the ITS region, PCR-RFLP, and SRAP markers. Twelve Morus species have been selected for this study from different region of Duhok. The ITS region's PCR result was 700 bp, but the matK gene's PCR produce was 900 bp. The same restriction site was found for all utilized cultivars when the 700bp of ITS fragment was used for PCR-RFLP with two restriction enzymes, RsaI GT/AC and HaeIII GG/CC. This study also used six combinations of SRAP markers to aid in grouping and identifying genetic similarities. The results of PCR-RFLP demonstrated an insufficient link between Morus physical appearance and genetic traits, but differences across studied cultivars could be identified using SRAP markers. Furthermore, this study demonstrated the possibility of DNA barcoding Morus cultivars, as well as additional sequence analysis and the identification of probable SNP between cultivars.

ABSTRACT During screening laboratory mice (Mus musculus) at the Zoology Department of the College of Science, King Saud University, some tiny oxyurid nematodes were detected from the large intestine of these rodents. Worms were identified using morphological and morphometric description. Furthermore, DNA was extracted from worms and subjected to polymerase chain reaction to amplify 18S rDNA and ITS (ITS1, 5.8S and ITS2) regions. Worms detected from caecum and colon of mice were detected in 80% of mice investigated. Morphologically, worms showed elongated body with tapered anterior end and narrow posterior part with cuticular annulations. Male worms measured 0.71-1.12 (0.90) long and 0.01-0.12 (0.11) wide. Female worms measured 3.12-5.011 (4.30) long and 0.13-0.29 (0.17) wide. Esophagus followed by intestine which opens at the posterior end via anal opening in females and via cloacal opening in males. Males have a single spicule with a gubernaculum and an accessory hook. Females’ uteri were pack with eggs. Data from 18S rDNA revealed a sequence which was identical to Syphacia obvelata in NCBI GenBank. Similarly, sequences from ITS regions grouped with sequences from S. obvelata confirming the morphological identity of the worm. However, it showed 3 mutations at the ITS2 region from related sequences from S. obvelata at NCBI GenBank.

Pedunculate oak is among the most economically important deciduous forest tree species in Europe and is also a host for many important ectomycorrhizal (ECM) fungi. The aim of this study was to analyse the ECM fungal community in a mature pedunculate oak stand near Morović, Serbia in spring and autumn. ECM fungi were determined by combining morpho-anatomical characterization of ectomycorrhizas with molecular analysis based on PCR amplification of the internal transcribed spacer region of fungal nuclear ribosomal DNA. The number of ECM fungal taxa and the number of different categories of fine roots were counted, diversity indices were calculated, and ECM fungi were classified into exploration types. Twenty-one ECM fungal taxa were recorded in the studied mature pedunculate oak stand, 19 in spring and 13 in autumn. ECM communities consisted of one dominant taxon and a larger number of rare taxa. Lactarius quietus was the most abundant ECM fungus in both seasons which made association with more than half of ECM root tips. At the stand near Morović, contact exploration type (ET) dominated, short-distance ET was less abundant, while medium-distance fringe ET and long-distance ET were rare in both seasons. The most pronounced difference between seasons is recorded in the number of ECM fungal taxa. The number of ECM fungal taxa and diversity indices recorded in the studied pedunculate oak stand were lower or similar compared to values obtained in stands of oak species across Europe.

Pomegranate is a tree species with a significant plant diversity, therefore molecular methods are necessary to define and verify approaches to recognize rapidly and correctly. This research looks at a genetic strategy for identifying pomegranate varieties in Duhok KRG. The approach is based on application of ITS region, PCR-RFLP, sequencing and SNP identification. For this study, 14 pomegranate accessions were taken from various regions, namely the Center of Duhok, Amedi, Akre, Zaxo, the South of Duhok, and Sulav. The PCR product of the 18S–28S rDNA intergenic spacer was 854bp, and the sequence analysis revealed a 99.94 percent similarity with other accession numbers in NCBI, demonstrating the use of the 18S–28S rDNA intergenic spacer for identifying and barcoding pomegranate cultivars. The PCR product of the ITS region was 700bp. This result was then employed for PCR-RFLP using two restriction enzymes namely RsaI GT/AC and HaeIII GG/CC which helped grouping as well as genetic similarities. This study Further involved sequencing examined genes were compared using the NCBI BLASTn tool and clustalo (Version 1.2.4) to determine gene location and SNP. According to this study, the result of PCR-RFLP revealed poor association between pomegranate physical morphology and genetic features, while SNP identification was identified between studied cultivars. Moreover, this result showed possible DNA barcoding of pomegranate cultivars under the study.

Microeukaryotes are vital for maintaining soil quality and ecosystem functioning, however, their communities are less studied than bacterial and fungal ones, especially by high throughput sequencing techniques. Alveolates are important members of soil microbial communities, being consumers and/or prey for other microorganisms. We studied alveolate diversity in soil under the undisturbed steppe (US) and cropped for wheat using two tillage practices (conventional, CT, and no-till, NT) by amplifying the ITS2 marker with ITS3_KYO2/ITS4 primers and sequencing amplicons using Illumina MiSeq. A total of 198 Alveolata OTUs were identified, with 158 OTUs attributed to the Ciliophora phylum, containing five classes: Litostomatea, Spirotrichea and Oligohymenophorea, Nassophorea and Phyllopharyngea. Litostomatea and Phyllopharyngea were more abundant in US as compared with CT and NT. The observed OTU richness was higher in US than in CT and NT. The β-biodiversity of soil ciliates also very distinctly differentiated the US field from CT and NT. In the US, Nassophorea and Spirotrichea correlated positively with sand and negatively with clay, silt and SOM contents. This is the first report about soil ciliates diversity in Siberia as assessed by metabarcoding technique. The revealed clear effect of land use on the relative abundance of some taxa and a lack of tillage effect suggest the importance of the quantity and quality of plant material input for shaping the prey for ciliates. The ITS-metabarcoding technique was used for the first time in the research of ciliates diversity; further studies, embracing diverse aspects of soil ciliates by combining -omics methodology with the traditional one, are needed to get a better insight on the ecological roles of the main ciliate taxa in the complex soil system.

Since Mulberries (Morus)is a tree species with a considerable plant variety. Molecular techniques are methods used to distinguish between species accurately, easily and quickly. This study examines a Molecular method for distinguishing different Morus species in the Duhok - Kurdistan region/Iraq. The method is based on the use of four techniques: matK gene, the ITS region, PCR-RFLP, and SRAP markers. Twelve Morus species have been selected for this study from different region of Duhok. The ITS region's PCR result was 700 bp, but the matK gene's PCR produce was 900 bp. The same restriction site was found for all utilized cultivars when the 700bp of ITS fragment was used for PCR-RFLP with two restriction enzymes, RsaI GT/AC and HaeIII GG/CC. This study also used six combinations of SRAP markers to aid in grouping and identifying genetic similarities. The results of PCRRFLP demonstrated an insufficient link between Morus physical appearance and genetic traits, but differences across studied cultivars could be identified using SRAP markers. Furthermore, this study demonstrated the possibility of DNA barcoding Morus cultivars, as well as additional sequence analysis and the identification of probable SNP between cultivars. [ABSTRACT FROM AUTHOR]

Simple Summary: Morphological and molecular data were collected from experimentally reared adult trematodes of Metorchis butoridi, a parasite of the gallbladder of fish-eating birds from the Russian Far East. Phylogenetic relationships within the family Opisthorchiidae were analyzed. The results confirmed that the trematodes under study belong to the genus Metorchis. Some taxonomical issues of the genus are discussed. Adult trematodes of the genus Metorchis were found in the gallbladders of ducklings that had been experimentally fed freshwater fishes of the genera Rhynchocypris and Rhodeus that were naturally infected by Metorchis metacercariae. Some of the trematodes were identified as Metorchis ussuriensis, whose morphology of developmental stages and molecular data had previously been described in detail. The other trematodes were confirmed as species Metorchis butoridi on the basis of morphological features: subterminal oral sucker, vitelline follicles with interrupted bands, and rosette-shaped testes. An analysis of phylogenetic relationships within Opisthorchiidae using nuclear and mitochondrial markers confirmed that the obtained trematodes were actually from the genus Metorchis. The morphological and molecular features indicated that a number of trematodes found in East Asia and described as Metorchis orientalis belong to M. butoridi. Also, the M. orientalis individuals from Europe are, in fact, representatives of another Metorchis species. [ABSTRACT FROM AUTHOR]

The endophytic fungi present in needles of Pinus sylvestris, Pinus pinaster, Pinus nigra, Pinus taeda and Pinus radiata were investigated in Kerpe Research Forest, İzmit in 2016. Ten trees of each pine species were sampled systematically. Previous years green needles were sampled from the lower part of the canopy, from two equally spaced positions around the tree. Each needle was surface sterilized and cut into 0.5 cm sections then individually placed onto malt extract agar plates. DNA was ex-tracted from reprensentative seven isolates and amplified using primers ITS1 and ITS4 targeting the nuclear 5.8S rDNA gene and the two ITS regions flanked between 18S and 28S rDNA genes. Amplicons sequenced in both directions using the univer-sal fungal primers ITS1 and ITS4. Isolations from a total of 1000 needles (200 from each pine species) yielded 750 fungal isolates. Pestalotiopsis funerea (Desm.) Steyaert was isolated at the highest frequency followed by Acremonium sp., Cladio-sporum sp. and Cyclaneusma minus.

The American cranberry is a perennial North American fruit plant that is grown successfully on commercial plantations in Poland. The purpose of this study was to recognize filamentous fungi that colonize roots, leaves, and fruits without visible disease symptoms. Pure fungal cultures were isolated from disinfected plant fragments in agar media and identified by sequencing common taxonomic DNA markers such as the ITS region, the TEF-1α, or RPB2 genes. Of the 141 isolates studied, 59% were identified as closely related to soil saprotrophs. They were classified primarily as showing the greatest similarity to type strains of Trichoderma amoenum, Trichoderma dorothopsis, Paraphaeosphaeria sporulosa, and Penicillium murcianum. Additionally, isolates that are most similar to strains of Penicillium crustosum, Aspergillus flavus, and Aspergillus versicolor that produced mycotoxins were detected. The fungi identified as closest to Alternaria geophila, Alternaria senecionicola, Paraphoma radicina, Pestalotiopsis unicolor, Pestalotiopsis scoparia, and Neopestalotiopsis spp., whose hosts are plants other than American cranberry, represented 33.81% of the isolates tested. Only 7.2% of the isolates corresponded to the species of Physalospora vaccinia, Diaporthe vaccinii, and Diaporthe eres, known cranberry pathogens. The results of this study can be used to identify latent plant infection and potential disease risks.

Fungus is the most important disease-causing biotic factor on earth. The study of their genetic diversity is very important for their management and control. For studying Ascochyta rabiei on chickpea blight chickpea samples caused by Ascochyta rabiei were collected and disease-causing organisms were isolated from infected plant parts and their ITS region was sequenced. BLAST analysis revealed that A. rabiei is the causal pathogen of this blight disease. Phylogenetic analysis suggested great diversity among these isolates of A. rabiei. The results confirm that A. rabiei strains collected from Bannu and Lakki Marwat have genetic similarities but are different from other isolated strains of the same species. Similarly, studied strains from Kamber, Shadat Kot, and Larkana. Khairpur and Bhakkar were also similar. However, strains from Khushab and Mianwali showed genetic differences from each other. The current research also reports the plant-mediated synthesis of ZnO NPs via the plant extract of Thevetia peruviana. The ZnO NPs were characterized through UV, XRD, FTIR, FE-SEM, EDX, and the size of the nanoparticle of 28 nm was calculated. The prepared nanoparticle was used for the antifungal activity against the most severe strain of A. rabiei. The Biosynthesis of ZnO NPs by plant assets is an eco-friendly, consistent process and appropriate for extensive production.

The present study, initially to resolve the cryptic species within Corollospora maritima, is to determine how to attain taxonomic discrimination at species and generic levels. Multiple sequence alignments (MSAs) of the ITS, 28S, and 18S regions of the nuclear ribosomal cistron were separately subjected to pairwise distance assessments, Bayesian, and Maximum likelihood phylogenetic analyses. Morphological descriptions of 15 type strains of Corollospora species, along with MSAs involving representatives of the whole genus Corollospora (268 isolates, many from C. maritima sensu lato) totaling 355 published sequences, allowed phylogenetic assessments conducted to the following p-distance thresholds in the ITS/28S regions: ≥3%/1% for species segregation and ≥8%/2% for generic segregation. This resulted in the introduction of 10 new genera encompassing 13 new combinations of current Corollospora species: Ajigaurospora pseudopulchella, Corollosporella anglusa, Corollosporella ramulosa, Corollosporopsis portsaidica, Garethelia parvula, Honshuriella fusca, Keraliethelia pulcehlla, Nakagariella filiformis, Paracorollospora angusta, Paracorollospora luteola, Paracorollospora marina, Shirahamella gracilis, and Tokuratelia colossa. Furthermore, seven undefined genera considered putative new genera (pNGenus A to G), and 16 undefined putative new species (seven spp. come from the resolution of the C. maritima complex), await re-assessment of their morphology and additional molecular data, which may result in the recognition of new taxa. [ABSTRACT FROM AUTHOR]

Reducing the use of fungicides, insecticides, and herbicides in order to limit environmental pollution and health risks for agricultural operators and consumers is one of the goals of European regulations. In fact, the European Commission developed a package of measures (the European Green Deal) to promote the sustainable use of natural resources and strengthen the resilience of European agri-food systems. As a consequence, new plant protection products, such as biostimulants, have been proposed as alternatives to agrochemicals. Their application in agroecosystems could potentially open new scenarios regarding the microbiota. In particular, the vineyard microbiota and the microbiota on the grape surface can be affected by biostimulants and lead to different wine features. The aim of this work was to assess the occurrence of a possible variation in the mycobiota due to the biostimulant application. Therefore, our attention has been focused on the yeast community of grape bunches from vines subjected to the phytostimulant BION®50WG treatment. This work was carried out in the CREA-VE experimental vineyard of Vitis vinifera cv. Barbera in Asti (Piedmont, Italy). The composition of fungal communities on grapes from three experimental conditions such as IPM (integrated pest management), IPM+BION®50WG, and IPM+water foliar nebulization was compared by a metabarcoding approach. Our results revealed the magnitude of alpha and beta diversity, and the microbial biodiversity index and specific fungal signatures were highlighted by comparing the abundance of yeast and filamentous fungi in IPM and BION®50WG treatments. No significant differences in the mycobiota of grapevines subjected to the three treatments were detected. [ABSTRACT FROM AUTHOR]

Endophytic fungi colonize the inner tissues and provide direct and indirect benefits to plants. Although Mongolia is rich in medicinal plants, due to climatic and anthropogenic reasons, the resources are being depleted, and many species are under threat of gradual extinction, while the endophytic fungi of Mongolian plants are largely unknown. In this study, a total of 24 culturable endophytic fungal strains were isolated from Cynomorium songaricum (Rupr.), a medicinal and vulnerable plant species of Mongolia. Based on the morphological characteristics and the sequences of the rDNA internal transcribed spacer (ITS) region, the isolates were identified into six genera: Fusarium (8), Clonostachys (7), Penicillium (6), Alternaria (1), Aspergillus (1), and Madurella (1). The antimicrobial activity was assessed by the agar-diffusion method, revealing that 15 strains were able to inhibit the growth of at least one of the test organisms. Among them, 1 strain showed inhibitory activity against Escherichia coli, 12 against Bacillus subtilis, 13 against Staphylococcus aureus, and 8 against Aspergillus niger, respectively. The ability to solubilize complex phosphorus and zinc minerals was observed in 3 and 21 strains, respectively, and the production of indole-3-acetic acid (IAA) was detected in nine strains in the presence of tryptophan. Our study provides the first insight into the cultivable endophytic fungal composition of C. songaricum, parasitizing the roots of Nitraria sibirica growing in the Gobi Desert of Mongolia. The resulting fungi, which have antimicrobial and plant growth-promoting properties, were preserved in the national culture collection and can be used to further exploit their biotechnological potential, as well as for the propagation of endangered and vulnerable medicinal plants.

Podocarpus is the most dominant genus of Podocarpaceae, with higher taxonomical proximity to the Taxaceae, having numerous pharmaceutical applications, however, scarce studies dealing with the physiological and metabolic criteria of Podocarpus in Egypt were reported. Thus, the objective of this work was to assess the physiological and metabolical patterns of the different species of Podocarpus; P. gracilior, P. elongates, P. macrophyllus and P. neriifolius. The highest terpenoids contents were reported in P. neriifolius, followed by P. elongatus, and P. macrophyllus. P. gracilior had the highest antioxidants amount, followed by P. macrophyllus, P. neriifolius and P. elongatus. From the GC/MS metabolic profiling, caryophyllene, β-cadinene, β-cuvebene, vitispirane, β-cadinene and amorphene were the most dominant metabolites in P. gracilior. β-Caryophyllene was the common in P. gracilior, P. elongatus, P. macrophyllus and P. neriifolius with an obvious fluctuation. The plant methanolic extracts have an obvious activity against the multidrug resistant bacteria; E. coli, P. aeruginosa, S. pyogenes and S. aureus, and fungi; A. fumigatus, A. flavus, A. niger and C. albicans in a concentration-dependent manner. The highest Taxol yield was assessed in the extracts of P. elongatus (16.4 μg/gdw), followed by P. macrophyllus, and P. neriifolius. The chemical identity of Taxol derived from P. elongatus was resolved by LC/MS, with molecular mass 854.6 m/z, and similar structural fragmentation pattern of the authentic one. The highest antitumor activity of P. elongatus extracted Taxol was assessed towards HCT-116 (30.2 μg/ml), HepG-2 (53.7 μg/ml) and MCF-7 (71.8 μg/ml). The ITS sequence of P. elongatus “as potent Taxol producer” was deposited on Genbank with accession #ON540734.1, that is the first record of Podocarpus species on Genbank.

Phlebopus portentosus is a popular wild edible mushroom that is found growing on both forest trees and fruit trees. Because it can grow either with various trees or without a host tree, it is reported that it is not a mycorrhizal fungus and may not be an obligate ectomycorrhizae. Furthermore, it was recognized that P. portentosus can be found in association with leguminous plants, such as Sesbania javanica Miq. The aims of this study were to cultivate P. portentosus with bioinoculants in solid and liquid forms and track the ectomycorrhizal association with Sesbania javanica Miq. A pure mycelium culture of this fungus was isolated, and the growth parameters, temperature, medium pH, and types of medium were examined. The mycelium grew very well on synthetic media such as MPDA and YM. The optimum pH and temperature were pH 4 and 30o C, respectively. Bioinoculants of P. portentosus were inoculated on seedbeds of Sesbania javanica Miq. After inoculation, ectomycorrhizal-like roots were found at 6 months, and sporocarps were found at 8.5 months. To prove this, the ITS rDNA sequences were aligned, and the results revealed that all sequences of the ectomycorrhizal-like root tips and sporocarps that developed on seedbeds of S. javanica Miq. and black bolete used as a bioinoculant were 100% identical to those of P. portentosus MN962534 (GenBank database). This finding indicates that P. portentosus can form ectomycorrhizae with S. javanica Miq. [ABSTRACT FROM AUTHOR]

This is the first report on the autumn-flowering species Crocus speciosus, belonging to C. ser. Speciosi from the Bulgarian flora. The species was found in Southeastern Bulgaria, in the area between Ahtopol and Rezovo. Re-examining the Bulgarian collections, the earliest specimen was collected in 1975, was probably overlooked, and most likely determined as C. pulchellus. The nearest known localities of the species are on the territory of Türkiye. In this study, we compared C. pulchellus and C. ibrahimii using DNA sequence data from the nuclear ITS1/2 region and morphological features. Our study showed a close relationship between the specimens from Bulgaria and the recently deposited data of C. speciosus and their separation from the closely related C. pulchellus. Together with the previously cited white anthers as a key feature for determination, the molecular data confirmed a clear distinction between the samples with white anthers in the two species. The morphological data of our taxon overlapped with the description of C. ibrahimii. The molecular data strongly supported the affiliation of C. speciosus s.l., but did not support the recognition of C. ibrahimii as a separate species and it should be referred to as a subspecies of C. speciosus subsp. ibrahimii Rukšāns. [ABSTRACT FROM AUTHOR]

Adult trematodes of the genus Metorchis were found in the gallbladders of ducklings that had been experimentally fed freshwater fishes of the genera Rhynchocypris and Rhodeus that were naturally infected by Metorchis metacercariae. Some of the trematodes were identified as Metorchis ussuriensis, whose morphology of developmental stages and molecular data had previously been described in detail. The other trematodes were confirmed as species Metorchis butoridi on the basis of morphological features: subterminal oral sucker, vitelline follicles with interrupted bands, and rosette-shaped testes. An analysis of phylogenetic relationships within Opisthorchiidae using nuclear and mitochondrial markers confirmed that the obtained trematodes were actually from the genus Metorchis. The morphological and molecular features indicated that a number of trematodes found in East Asia and described as Metorchis orientalis belong to M. butoridi. Also, the M. orientalis individuals from Europe are, in fact, representatives of another Metorchis species.

Objectives: Endophytic fungi are widely known to reside in plant tissues causing no harm to host plants or even no noticeable change. They may colonize host plants for a part of their life cycle or may complete the entire life cycle since host provide a variety of novel metabolites. Despite being in a close relationship with medicinal host plants, the diversity and metabolic spectrum of endophytic fungi have not been researched well. Their diversity and ecological niche as plant symbionts make them attractive targets in the search for novel biochemicals. Methods: We aimed to isolate, purify, and characterize the endophytic fungal population of medicinal host plants cultivated in Riyadh, Saudi Arabia. The goal was achieved through morphological identification and internal transcribed spacer (ITS) regions in rRNA analyses to determine phylogeny and diversity. Three aromatic and medicinal plants Jasminum sambac, Camellia sinensis, and Ocimum basilicum collected from the Riyadh region, Saudi Arabia were investigated. In total, 84 isolates were purified and they were grouped into 20 operational taxonomic units (OTUs) as per their sequences of ITS regions in rRNA. Results: Twenty species of endophytic fungi were grouped in 12 genera i.e. Neopestalotiopsis, Trichoderma, Fusarium, Colletotrichum, Myrothecium, Chaetomium, Alternaria, Phoma, Curvularia, Cladosporium, Neodidymelliopsis, and Aspergillus and all isolates belonged to Ascomycota phylum. J. sambac was found dominant among other and had a relative frequency of 27%. C. sinensis was next with 18.7% relative frequency. The diversity was prominently recorded in leaf organs over stem and roots while roots exhibited the lowest diversity. Isolates also produced indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzyme, and siderophores with variable magnitudes that could be assigned to their wide range of metabolic variations from species to species. Conclusion: Conclusively, J. sambac, C. sinensis, and O. basilicum are a good source of endophytic fungi with certain plant growth-promoting traits. Overall, Alternaria was found as the most predominant genus in terms of colonization rate. Further deternminations are required to screen the beneficial compounds released by these endophytic fungi.

Kiwi is one of the best natural sources of vitamin C and has wide applications. During October–November 2021, small brown spots were examined on the Kiwi leaves. The diseased leaf samples were collected and placed on potato dextrose agar nutrient media for diagnosis. Morphological, anatomical, and molecular studies revealed this disease-causing agent to be Rhizopus oryzae. Molecular characterizations of the isolated pathogen were performed by using actin translation elongation factor (EF-1α) and ribosomal deoxyribose nucleotide inter transcribed sequence (rDNA ITS ITS1/ITS4) and elongation factors (EFl-F/EFl-R) primers. A BLAST study of the resultant ITS1/ITS4 sequence showed > 99% resemblance with R. oryzae (MT603964.1), while the EF-1α sequence revealed 100% similarity with translation elongation factor-1α gene of R. oryzae (MK510718.1). The obtained ITS1/ITS4 sequence was submitted to NCBI (MW603842.1). Koch's postulates established the pathogenicity of isolated R. oryzae and proved it to be the brown spot pathogen of Kiwi. For the environmentally-friendly management of Kiwi leaf spot, maize biochar-Zinc Oxide (MB-ZnO) nanocomposite was used. The prepared nanocomposite was characterized by Fourier transform infrared (FTIR) spectroscopy, thermo gravitational analysis (TGA), X-ray diffraction (XRD) analysis, scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) analysis. After successful preparation, MB-ZnO was assessed for its possible antifungal potential against R. oryzae. MB-ZnO displayed substantial growth inhibition, and the highest growth inhibition (79%) was observed at a 19 mg/mL dose rate of nanoparticles. These excellent findings propose that Ball-milled synthesis is a fast, economical, and environmentally friendly method for nanocomposite in the near future. The nanocomposite is used as a nominal substitute for chemical fungicides. [ABSTRACT FROM AUTHOR]

Protea (Protea cynaroides L.) is one of the most important ornamental plants commercially cultivated in Turkey and other parts of the world. In May 2017, protea plants heavily showed wilting symptoms were obtained from the protea growing areas at Hatay province of Turkey. Brown vascular tissue in the stems of the infected plants was observed. To determine the causal agent of the disease, necrotic tissue pieces taken from diseased plants were surface-sterilized with 10% sodium hypochlorite, two times rinsed with sterile distilled water, placed on Petri dishes containing potato dextrose agar (PDA), and incubated at 24 oC for seven days. Microconidia were no septate, ovoid, hyaline, single-celled, and 9 to 12 × 1 to 3 μm. Macroconidia were slightly curved, four to six septate, and 27 to 31 × 3 to 5 μm. Based on the morphology and cultural features, the fungus was identified as F. incarnatum, which belongs to the F. incarnatum-equiseti species complex (FIESC). In addition, a pathogenicity test was made using the apple fruits. The pathogen was re-isolated from inoculated apple fruits and both cultural and morphological characteristics of the pathogen were identical. To confirm the identity of one representative isolate, the internal transcribed spacer (ITS) region including 5.8S rDNA was amplified and sequenced with primers of ITS-1 and ITS-4. Fusarium MLST, Fusarium-ID and GenBank database were used for the identification of sequence. The amplified 465 bp product has 99% nucleotide identity with the sequences of F. incarnatum-equiseti species complex of MLST types and deposited in GenBank under accession number (MH005097). According to the literature, this is the first molecular identification of F. incarnatum on protea plants in Turkey.

Carpinus is a genus of the family Betulaceae that comprises 42 species worldwide. Moreover, more than a third of total Caspian forests are occupied by C. betulus and it has an important role in slope stabilizing. C. orientalis, commonly known as oriental hornbeam, is a small tree or often shrub, rarely over 10 m tall, and a major pioneer species on slopes in shallow humus-poor or rocky soils. This species is distributed from southeastern Europe to the north of Iran, from west to easternmost of the Hyrcanian forest. Due to their peculiar and beautiful fruit cluster, some hornbeams are used as important ornamental plants. Taxonomy of this genus has always been problematic in Iran and the number of species ranges from 2 to 4 in different taxonomic literature. In the current study, we applied morphometric (PCA and cluster analyses) and molecular (ITS region) approaches to delineate the species boundary of the genus in Iran. Thirty-six quantitative and qualitative characters were used for morphological analyses. The PCA plot of morphological data divided the studied population into three groups. However, the cluster analysis revealed two major groups. Moreover, Iranian species of the genus Carpinus formed two distinct clades in the molecular analyses. The results of the present study showed that there are two Carpinus species in Iran, including C. betulus and C. orientalis with two subspecies and C. schuschaensis is introduced as a synonym for C. orientalis subsp. macrocarpa. In addition, the intraspecific morphological diversity has blurred species boundaries.

The present study, initially to resolve the cryptic species within Corollospora maritima, is to determine how to attain taxonomic discrimination at species and generic levels. Multiple sequence alignments (MSAs) of the ITS, 28S, and 18S regions of the nuclear ribosomal cistron were separately subjected to pairwise distance assessments, Bayesian, and Maximum likelihood phylogenetic analyses. Morphological descriptions of 15 type strains of Corollospora species, along with MSAs involving representatives of the whole genus Corollospora (268 isolates, many from C. maritima sensu lato) totaling 355 published sequences, allowed phylogenetic assessments conducted to the following p-distance thresholds in the ITS/28S regions: ≥3%/1% for species segregation and ≥8%/2% for generic segregation. This resulted in the introduction of 10 new genera encompassing 13 new combinations of current Corollospora species: Ajigaurospora pseudopulchella, Corollosporella anglusa, Corollosporella ramulosa, Corollosporopsis portsaidica, Garethelia parvula, Honshuriella fusca, Keraliethelia pulcehlla, Nakagariella filiformis, Paracorollospora angusta, Paracorollospora luteola, Paracorollospora marina, Shirahamella gracilis, and Tokuratelia colossa. Furthermore, seven undefined genera considered putative new genera (pNGenus A to G), and 16 undefined putative new species (seven spp. come from the resolution of the C. maritima complex), await re-assessment of their morphology and additional molecular data, which may result in the recognition of new taxa.

Although oaks belong to the economically most important hardwood tree species in Europe, data on the diversity of ectomycorrhizal (ECM) fungi on pedunculate oak in the Republic of Serbia are deficient. The aim of our study was to give the first insight into the diversity of ECM fungi in the young stand of pedunculate oak in Morović, Serbia. The combination of morpho-anatomical characterization and molecular analysis was used for determination of ECM fungi on penduculate oak. Studied parameters included: (i) counting of vital ECM root tips, (ii) calculation of diversity indices and (iii) classification of morphotypes of ectomycorrhizae into the exploration types. Eighteen ECM fungal taxa were recorded in the studied young pedunculate oak stand. Seven of them were identified to the level of species, eight fungi to the genus, two to the family level, and one ECM fungus remained unidentified. ECM communities consisted of a small number of abundant taxa and a bigger number of rare taxa. The most abundant ECM fungi were Entoloma sp., Thelephoraceae sp., Russula cf. odorata and Russula lilacea, which made association with the majority of ECM root tips. Short-distance exploration type (ET) dominated, followed by medium-distance smooth ET and contact ET, while long-distance ET and medium-distance fringe ET were rare at the Morović site. Values of diversity indices obtained in the studied pedunculate oak stand were comparable to the ones recorded in different oak stands over Europe. In order to get a deeper insight into the diversity of ECM fungi on pedunculate oak there is a need to continue research on increased number of sites and also to include seasonal dynamics.

The classification of Thlaspi s.l. is still problematic. Earlier phylogenetic research of the genus focused on several small groups within Thlaspi s.str. and lacked detailed morphological observations. The relationships among Eurasian taxa and the value of fruit morphology in defining them have yet to be studied. The aim of this study was to analyze 22 taxa belonging to the Nomisma, Thlaspi and Pterotropis sections of Thlaspi s.l. from Turkey using maximum likelihood (ML) analysis of internal transcribed spacer (ITS) sequences. We also analyzed the morphological features of the fruit. According to the results, the examined taxa fell into 2 main clades. Moreover, clade II showed 3 sub-clusters. Thlaspi huetii and T. aghricum were the most distant taxa with a distance of 0.49%; however, T. ochroleucum and T. violascens were found to be 99% similar. According to ITS region data based on multiple populations of each taxon, T. arvense, T. huetii, T. perfoliatum, T. violascens, T. cataonicum, T. elegans, T. rosulare and T. aghricum were placed together in one cluster, which indicates that they are monophyletic. Thlaspi elegans was found to be a polyploid complex based on bootstrap (BS) (a resampling technique that uses replacement sampling to estimate statistics in a population) values, which varied widely among the studied T. elegans taxa (98, 65 and 49%). Fruit morphology also supported the inter-specific relationships based on molecular data, and relationships found by ITS region data were compatible with fruit type and geographic distribution. A diagnostic key based on fruit morphology is provided for the identification of the examined Thlaspi taxa. [ABSTRACT FROM AUTHOR]

The identification and classification of strains of Acanthamoeba, a potentially pathogenic ubiquitous free-living amoeba, are largely based on the analysis of 18S rDNA sequences, currently delineating 23 genotypes, T1 to T23. In this study, the sequences of the ITS region, i.e., the 5.8S rDNA and the two internal transcribed spacers (ITS-1 and ITS-2), and those of the large subunit (LSU) rDNA of Acanthamoeba were recovered from amoeba genomes; the sequences are available in GenBank. The complete ITS–LSU sequences could be obtained for 15 strains belonging to 7 distinct lineages (T4A, T4D, T4F, T4G, T2, T5, and T18), and the site of the hidden break producing the 26Sα and 26Sβ was identified. For the other lines, either the LSU is partial (T2/T6, T7) or the ITS is fragmentary (T7, T10, T22). It is noteworthy that a number of sequences assigned to fungi turned out to actually be Acanthamoeba, only some of which could be affiliated with known genotypes. Analysis of the obtained sequences indicates that both ITS and LSU are promising for diagnostic and phylogenetic purposes. [ABSTRACT FROM AUTHOR]

(1) Background: Species of the anamorphic genus Cladobotryum, are known for their fungicolous lifestyle, making them important mycopathogens in fungiculture. Many morphological, ecological, and molecular phylogenetic studies of the genus have been done to date, but taxonomic uncertainties and challenges still remain. Fungal secondary metabolites, being vastly diverse, are utilised as an extra tool in fungal systematics. Despite being studied for their potentially bioactive compounds, Cladobotryum species are insufficiently investigated regarding metabolomics. (2) Methods: The aim of this study is the identification of Greek strains of Cladobotryum by integrating morphological data, ITS-based phylogeny, and 1H NMR-based metabolomics into a polyphasic approach. (3) Results: Twenty-three strains, isolated from sporophores of macromycetes inhabiting diverse Greek ecosystems, were morphologically identified as Cladobotryum apiculatum, C. fungicola, C. mycophilum, C. varium, C. verticillatum, and Hypomyces rosellus (anamorph C. dendroides), whereas seven strains, which produced red-pigmented metabolites, presented an ambiguous taxonomic position at the species level. Molecular phylogenetics and metabolomics corroborated the morphological findings. (4) Conclusions: Thorough morphological study, ITS region-based phylogeny, and NMR-based metabolomics contribute complementarily to resolving the genus Cladobotryum systematics. [ABSTRACT FROM AUTHOR]

Managing soil biodiversity by reduced or no tillage is an increasingly popular approach. Soil mycobiome in Siberian agroecosystems has been scarcely studied; little is known about its changes due to tillage. We studied mycobiome in Chernozem under natural steppe vegetation and cropped for wheat by conventional or no tillage in a long-term field trial in West Siberia, Russia, by using ITS2 rDNA gene marker (Illumina MiSeq sequencing). Half of the identified OTUs were Ascomycota with 82% of the total number of sequence reads and showing, like other phyla (Basidiomycota, Zygomycota, Mortierellomycota, Chytridiomycota, Glomeromycota), field-related differential abundance. Several dominant genera (Mortierella, Chaetomium, Clonostachys, Gibberella, Fusarium, and Hypocrea) had increased abundance in both cropped soils as compared with the undisturbed one and therefore can be safely assumed to be associated with wheat residues. Fungal OTUs' richness in cropped soils was less than in the undisturbed one; however, no tillage shifted soil mycobiome composition closer to the latter, albeit, it was still similar to the ploughed soil, despite different organic matter and wheat residue content. The study provided the first inventory of soil mycobiome under different tillage treatments in the south of West Siberia, where wheat production is an important section of the regional economy. [ABSTRACT FROM AUTHOR]

In recent years, new data on the diversity of genera and species in the phylum Glomeromycota continue to be added and rearranged. Arbuscular mycorrhizal fungi are key to plant nutrition and agriculture. Studies report different short- and long-term cultivation practices that influence the abundance and diversity of Glomeromycota. To the best of our knowledge, there are no known studies of the fungal communities in the fine aroma cocoa cultivars. In this context, our work aims to discover the diversity of arbuscular mycorrhizae associated with two cocoa cultivation practices (conservative and semi-conservative) through the isolation of spores using microscopy and metabarcoding of the internal transcribed spacer region (ITS). Morphological analysis showed that the density of Glomeromycota spores exhibited significant differences between production systems. Although the metabarcoding analysis showed that diversity indices showed a higher increase in the roots than in the cocoa soil, independently of the cultivation practice. An abundance of 348 and 114 taxa were observed, corresponding to the conservative and semi-conservative practices, respectively. Seven genera were observed for the first time in cocoa crop agroforestry systems, including P. scintillans, R. diaphanus, R. fasciculatus, R. custos, D. disticha, M. perpusilla, and D. bernensis. [ABSTRACT FROM AUTHOR]

Several nematode species can be found in different densities in almost any soil ecosystem, and their diversity in those ecosystems depends on numerous reasons, such as climatic conditions and host presence. Cereals are one of the main hosts of plant-parasitic nematodes (PPN), chiefly root-lesion nematodes (RLN, Pratylenchus spp.) and cereal cyst nematodes (CCN, Heterodera spp.). These nematodes are known as major parasites of the cereal crops; however, agricultural areas accommodate various nematodes showing biological variation. The diversity of parasitic nematodes on cereals in the Sakarya provinces of Türkiye, where cereals are intensively grown and located in the middle of two climatic zones, has not been well studied. Therefore, in this study, we aimed to determine the diversity, identification, and molecular phylogeny of PPNs in wheat-growing ecosystems in the Hendek, Pamukova, Geyve, Akyazı, and Central districts of Sakarya. The diversity of PPNs was calculated using the Shannon diversity index. Thirteen PPN genera were detected in 92% of soil samples. Heterodera filipjevi was identified in 24% of the soil samples using morphological, morphometrical, and molecular tools. In the morphological and molecular analyses, intraspecific polymorphism was observed in H. filipjevi populations. The result indicated that the high infestation rate of H. filipjevi was recorded from Geyve and Pamukova, followed by Hendek and Akyazı; however, a low infestation rate was detected in the Central district. The moderate value of the Shannon index of migratory nematode species was obtained in wheat fields as 2.31, whereas the value of evenness was 0.93, implying moderate diversity and high evenness of nematodes. This study is the first comprehensive report on H. filipjevi from wheat cropping areas in the Sakarya province. Intensified cereal cropping systems with/without non-cereal rotations increased the risk of plant-parasitic nematodes, especially RLNs and H. filipjevi infection of wheat production areas in the province. [ABSTRACT FROM AUTHOR]

Myxobolus nagaraensis is a myxozoan parasite first reported in freshwater gobies ( Rhinogobius spp.) from the Nagara River, Gifu Prefecture, Japan. Myxospores of M. nagaraensis form plasmodia in the visceral cavities of gobies, commonly presenting as distended abdomens. Although Rhinogobius is a common fish genus in Japan, details of M. nagaraensis , including genetic information, remain unknown. We compared the nucleotide sequences of the ribosomal RNA gene (rDNA) of M. nagaraensis from three different host species ( R. fluviatilis , R. nagoyae , and R. similis ) caught in three different rivers in Japan (Sakai, Sagami, and Kaname). The ITS region (ITS-1, 5.8S rDNA, and ITS-2) and large subunit (LSU) rDNA exhibited 49 and 55 variable sites, respectively. The highest nucleotide diversity was observed in the ITS region (0.00962), whereas that of the LSU rDNA was 0.00187. Differences in host species, rather than rivers, were a significant factor for genetic variation in both the ITS region (62.58%; P  < 0.001) and LSU rDNA (55.22%; P  < 0.01). Significant genetic variation was observed in M. nagaraensis from R. similis compared to R. fluviatilis ( P  < 0.001) or R. nagoyae ( P  < 0.001) from the same river. Such details are valuable for understanding parasite dispersal and its ecological impact on Rhinogobius hosts., Competing Interests: The authors declare that there are no known conflicts of interest associated with this publication., (© 2024 The Authors. Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Soybean (Glycine max L.) is one of the most valuable oilseed crops in the world. It is not only an oil seed crop and feed for livestock, but also valuable mineral and vitamins sources for the human diet. The soybean yield is affected by various biotic and abiotic stress factors in all growing seasons. Diseases are one of the most significant biotic factors that reduce soybean growth and yield. Fungi are important pathogens affecting yield and quality by attacking plants during the growth period and after harvest. This study was conducted to detect and identify the seed-borne fungi associated with the soybean seed. From this context, 150 soybean seeds were randomly chosen from the experimental fields of Akdeniz University in Antalya province of Turkey. These seeds were sterilized with 70% ethanol for 1 min, followed by 10% sodium hypochlorite for 1 min and then rinsed with sterile water and then placed in Petri plates by using the agar plate method. A total of four seedborne fungi species namely Aspergillus spp., Penicillium spp., Cladosporium spp. and Fusarium spp. were isolated from the soybean seeds. Additionally, Genomic DNAs of these fungal species were extracted and the internal transcribed spacer (ITS) region of ribosomal DNA was amplified with the ITS-1 and ITS-4 primers using a thermal cycler. After sequencing of amplified products, the sequences were aligned. BLASTn analysis of each sequence showed that the sequences of the fungi had the similarity (99%) to the fungal isolates deposited in the GenBank.

Abstract Background Spodoptera litura Fabricius (Lepidoptera: Noctuidae) is an important pest causing severe damage to many cultivating plants such as corn worldwide, including Indonesia. This study was performed to obtain and identify entomopathogenic fungi (EPF) of S. litura collected from corn fields in 4 corn producing regions of Lampung, Indonesia, as well as to investigate the damage caused by this pest on corn in Lampung Province. Results Three corn fields in each region were selected for collecting soil samples. Soil samples were collected from 5 corn plant rhizospheres, at each field in six months of survey. Ten larvae of Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae) were laid on each soil sample as a bait, covered with a filter paper and incubated at room temperature. The emerging fungi from T. molitor cadaver were transferred onto Potato Dextrose Agar (PDA) medium and incubated for 7 days at room temperature. Pathogenicity test was determined against 3rd instar of S. litura larvae. Identification was performed based on the sequence of Internal Transcribed Spacer (ITS) Region. Observations on the corn damage caused by S. litura were conducted at all corn producing areas in Lampung. Twelve fungal isolates were obtained causing 0–75% of mortality of S. litura. Four fungal isolates (NKPT, SKHJ, SDHJ and RAHJ), which caused mortality more than 20%, were further identified. One isolate (NKPT) was confirmed as Beauveria bassiana and the other 3 isolates (SKHJ, SDHJ and RAHJ) were Aspergillus oryzae. S. litura generally caused slight damages to the corn which was found in every observation year performed during 2010–2019. Medium plant damage was observed in 2010–2012 and 2018–2019, severe damage was found in 2011 and crop failure was recorded in 2018. Conclusions Aspergillus oryzae and B. bassiana were the EPF recorded infecting S. litura in corn in Lampung Province. This was the first report on the isolates of A. oryzae as EPF of S. litura in Indonesia. Slight damages with S. litura were always recorded in every observation year but not for those of medium and severe damages and crop failure.

Aims: Molecular identification of yeast has been conducted on various fermentation products. However, the identification of yeast in fermented Sumbawa mare's milk based on the genotyping method has not been carried out. This study was aimed to determine the diversity profile of yeasts in fermented Sumbawa mare's milk using phenetic characters and PCR-RFLP analysis technique based on the ITS region. Methodology and results: Yeast isolates were phenotypically characterized and visualized in a dendrogram using CLAD97 software. Then, the yeast DNA was extracted using heat treatment and amplified using ITS1 and ITS4 primers. The amplicons were analyzed by RFLP using HindIII and HaeIII enzymes. The phylogenetic tree was constructed using MEGA 7.0. Based on the result of grouping by phenetic analysis and PCR-RFLP, the 12 isolates were divided into four groups with different members. The results of the phenetic analysis were divided into group I (all isolates of Dompu), group II (isolate B3, B4, S3), group III (isolate B5) and group IV (isolate S1). The types of yeast that were identified molecularly and represented each group of PCR-RFLP results included in group I were Kluyveromyces marxianus D1A and K. marxianus D1B, group II: K. marxianus D7, group III: Kazachstania humilis D4, while milk from Bima and Sumbawa has one yeast species as a member of group IV, namely Pichia kudriavzevii B3. Kluyveromyces marxianus was the yeast frequently found in Sumbawa fermented mare's milk. Conclusion, significance and impact of study: Various yeast species as a consortium of the milk samples can contribute to the increasing quality of fermented Sumbawa mare's milk. [ABSTRACT FROM AUTHOR]

Health concerns and financial losses caused by mushroom poisoning have been reported worldwide. Amanita citrinoannulata, a poisonous mushroom commonly found in China, results in a toxic reaction in humans after mistaken ingestion. To reduce the mistaken ingestion of poisonous mushrooms and to improve clinical diagnosis of mushroom poisoning, a rapid mushroom species identification method is required. Such identification methods could be advantageous in the identification of other poisonous mushroom species. This study developed two rapid and sensitive methods for the detection of A. citrinoannulata utilizing colorimetric and real-time loop-mediated isothermal amplification (LAMP) technology and specifically designed primers for the internal transcribed spacer (ITS) genes of A. citrinoannulata. The methods demonstrated high sensitivity as 0.2 ng of A. citrinoannulata DNA could be detected, with no cross-reaction with 41 non-target mushroom species. The entire detection process could be completed within 40 min without requiring complex instruments and can be observed by the naked eye. Therefore, these novel methods can be used for the identification of fresh and cooked mushroom samples and vomit samples, which contain only 1% A. citrinoannulata. Furthermore, these methods facilitate the detection of mushroom poisoning, and thus, have potential to reduce the number of mushroom poisoning-related deaths worldwide.

Species from the family Kareniaceae (Dinophyceae) frequently cause harmful algal blooms (HABs), with serious ecological impacts and risks to human safety and aquaculture activities in coastal waters worldwide. However, due to their small size, lack of morphological divergence, and low abundance during non-bloom periods, the diversity within this family is not well understood. By comparing the commonly used molecular markers, the Internal Transcribed Spacer (ITS) region was found to have an appropriate mutation rate to distinguish three of the most common genera (Karenia, Karlodinium, and Takayama) within the Kareniaceae family and different geographical strains of Kareniaceae. Specific primers targeting the ITS region of Karenia, and the other primers specific to the genera Karlodinium and Takayama, were designed. Specificity of the primers was tested using 17 strains of Kareniaceae species and 15 non-target species. Representative Kareniaceae species could be successfully detected even at low concentrations of target DNA template with a limit of detection of 3.2 pg. The primers were also assessed using high-throughput sequencing with two environmental samples from the South China Sea (SCS). Analysis of the reads identified as Kareniaceae species revealed a high diversity and the existence of unreported Kareniaceae species in the SCS. In conclusion, the newly developed molecular barcodes specifically detected Kareniaceae species in the field and will provide technical support for the effective warning and monitoring of Kareniaceae HABs.

Abstract Background Banana fruit scarring beetle (BFSB), Basilepta subcostata (Jac.) (Chrysomelidae:Coleoptera), is an important insect pest feeds on leaf and fingers, which affects the cosmetic value of the fruit. The pest is distributed in Assam, Bihar, West Bengal, Chhattisgarh, and North-eastern Hill regions of India. Results The pest is currently managed by foliar spray with insecticides. In order to identify eco-friendly control of the pest, attempts were made to isolate microbial agent and evaluate their potential to control the pest. A total of 27 entomopathogenic fugal isolates were obtained from Odoiporus longicollis (Oliver), Cosmopolites sordidus (Germar), Basilepta subcostata (Jac), and other insect Galleria mellonella (Fabr). Based on colony morphology, the collected fungal isolates were identified as Metarhizium spp. (17) and Beauveria spp. (10). Through ITS sequencing, the fungal isolates were further characterized at species level as B. bassiana (8), B. brongniartii (2), M. anisopliae (8), M. robertsii (6), M. guizhouense (2), and M. pinghaense (1). Their sequences were submitted in GenBank and obtained accession numbers. Among 27 isolates tested against B. subcostata under laboratory conditions, 3 isolates (M. anisopliae NRCBEPF-36, M. pinghaense NRCBEPF-7 and B. brongniartii NRCBEPF-27) recorded 100% beetle mortality, followed by 11 isolates with 95–99% and 13 isolates with 88–93% within 8 days of treatment. Conclusion This study highlights the two native North East India isolates B. brongniartii NRCBEPF-27 (MT151781) and M. anisopliae NRCBEPF-36 (MT140308) showed the significance to use as potential biocontrol agents against banana fruit scarring beetle B. subcostata. Further experiments under field conditions are required to evaluate their biological control efficacy against the pest.

This is the first report on the autumn-flowering species Crocus speciosus, belonging to C. ser. Speciosi from the Bulgarian flora. The species was found in Southeastern Bulgaria, in the area between Ahtopol and Rezovo. Re-examining the Bulgarian collections, the earliest specimen was collected in 1975, was probably overlooked, and most likely determined as C. pulchellus. The nearest known localities of the species are on the territory of Türkiye. In this study, we compared C. pulchellus and C. ibrahimii using DNA sequence data from the nuclear ITS1/2 region and morphological features. Our study showed a close relationship between the specimens from Bulgaria and the recently deposited data of C. speciosus and their separation from the closely related C. pulchellus. Together with the previously cited white anthers as a key feature for determination, the molecular data confirmed a clear distinction between the samples with white anthers in the two species. The morphological data of our taxon overlapped with the description of C. ibrahimii. The molecular data strongly supported the affiliation of C. speciosus s.l., but did not support the recognition of C. ibrahimii as a separate species and it should be referred to as a subspecies of C. speciosus subsp. ibrahimii Rukšāns.

The paper presents the results of phytopathogenic fungi determination in bare-root forest nurseries of Middle Siberia. Genetic analysis of pathogenic microflora of Pinus sylvestris L., Pinus sibirica Du Tour and Picea obovata Ledeb. seedlings allowed identification of 17 genera of micromycetes: Phoma Sacc., Lophodermium Chevall., Sclerophoma Höhn. (teleomorph Sydowia Bres.), Cladosporium Link, Alternaria Nees, Typhula (Pers.) Fr. etc. Most frequently detected fungi represented genera Phoma (23.7 %) and Lophodermium (23.6 %). Pinus sylvestris L. seedlings harboured the highest diversity of fungal taxa. Seven genera of microscopic fungi (Phoma sp., Didymella sp., Alternaria sp., Cladosporium sp., Lophodermium sp., Gremmenia sp., Sclerophoma sp.) were detected in all studied forest zones: taiga, forest-steppe and Southern-Siberian mountain. The obtained results demonstrate the usefulness of DNA analysis for the identification of phytopathogenic fungi in forest nurseries of Middle Siberia with several implications for increasing the efficacy of forest management.

Protea (Protea cynaroides L.) is one of the most important ornamental plants commercially cultivated in Turkey and other parts of the world. In May 2017, protea plants heavily showed wilting symptoms were obtained from the protea growing areas at Hatay province of Turkey. Brown vascular tissue in the stems of the infected plants was observed. To determine the causal agent of the disease, necrotic tissue pieces taken from diseased plants were surface-sterilized with 10% sodium hypochlorite, two times rinsed with sterile distilled water, placed on Petri dishes containing potato dextrose agar (PDA), and incubated at 24 oC for seven days. Microconidia were no septate, ovoid, hyaline, single-celled, and 9 to 12 × 1 to 3 µm. Macroconidia were slightly curved, four to six septate, and 27 to 31 × 3 to 5 µm. Based on the morphology and cultural features, the fungus was identified as F. incarnatum, which belongs to the F. incarnatum-equiseti species complex (FIESC). In addition, a pathogenicity test was made using the apple fruits. The pathogen was re-isolated from inoculated apple fruits and both cultural and morphological characteristics of the pathogen were identical. To confirm the identity of one representative isolate, the internal transcribed spacer (ITS) region including 5.8S rDNA was amplified and sequenced with primers of ITS-1 and ITS-4. Fusarium MLST, Fusarium-ID and GenBank database were used for the identification of sequence. The amplified 465 bp product has 99% nucleotide identity with the sequences of F. incarnatum-equiseti species complex of MLST types and deposited in GenBank under accession number (MH005097). According to the literature, this is the first molecular identification of F. incarnatum on protea plants in Turkey. [ABSTRACT FROM AUTHOR]

Plant identification is an overwhelming task due to different biological attributes and great diversity in plant species. In the absence of physical markers, molecular techniques have become useful for the identification of species of origin of medicinal plant seeds, pastes, and formulations of suspected plants. The ITS region of nuclear rRNA was amplified from thirteen different toxic plant species by using universal primer ITS 1 & 4. Nucleotide sequences of all selected plants were submitted in NCBI and accession numbers were acquired. The results of this study give accurate identification of thirteen plant species and proved the ITS region of 18s-26s nuclear ribosome to be an important tool for phylogenetic analysis and species identification of plants. The sequence was aligned with top matched reference sequence and presented in Clustal Omega software for making a phylogenetic neighbour tree. The significance of these findings is paramount in forensic toxicology scenarios especially when fragmentary plant material is found in the stomach/intestine and its morphological identification becomes impossible. In these circumstances, the PCR based molecular technique surely plays a significant role in solving complicated forensic cases.

Abstract Background Enterocytozoon bieneusi, a zoonotic pathogen, has the potential to infect both immunocompromised and immunocompetent humans. It is found in large number of animals; however, not much is known regarding its prevalence in equine animals, particularly donkeys. This is the first molecular epidemiological evaluation of E. bieneusi in 178 free-ranging donkeys from five countrysides; and 502 farmed donkeys from 18 farms in 12 cities of Xinjiang, China by Nested PCR. Results E. bieneusi was detected in 2.5% (17/680) donkeys, with 2.6% (13/502) in farmed and 2.2% (4/178) in free-ranging ones. Sequence analysis identified eight ITS genotypes, all belonging to zoonotic Groups 1 or 2, including six known genotypes: horse1 (n = 5), D (n = 3), NCD-2 (n = 3), BEB6 (n = 2), BEB4 (n = 1), and NIAI (n = 1); and two new genotypes: XJD1 (n = 1) and XJD2 (n = 1). Conclusions This is the first report confirming the presence of E. bieneusi in donkeys in Xinjiang, China, and indicates the possibility of zoonotic transmission of this pathogenic parasite.

Verticillium spp., including V. nonalfalfae and V. dahliae, are known vascular wilt pathogens of the invasive Ailanthus altissima (tree-of-heaven) in the United States and in Europe. Herein we provide evidence of the presence of a previously unreported wilt disease of A. altissima in Tuscany (Central Italy). Several isolates were collected from two locations and identified as V. dahliae, based on microscopical features of conidiophores, conidia and microsclerotia. Genomic DNA was extracted from the mycelium, the ITS region was amplified and the sequence was deposited in GenBank as VdGL16 (accession no. MK474459). BLASTn analysis showed 100% similarity with V. dahliae. To confirm pathogenicity of VdGL16, inoculations of Ailanthus seedlings were performed with the root dipping technique whereas mature trees were stem-inoculated. All inoculated seedlings exhibited wilt symptoms after 20 days, while mature Ailanthus trees showed wilting and dieback after six months. The pathogen was easily re-isolated from seedlings and re-identified as V. dahliae, thus satisfying Koch’s postulates. Results from intraspecific resistance screening of nine seed sources from across Italy revealed that Ailanthus provenances from all the six sampled regions were susceptible to V. dahliae. Stem inoculated adult plants exhibited abundant production of epicormic sprouts along the stem within six months, and most of these sprouts wilted following initial dieback of the main stem; furthermore, sprouting from the crown was intense. Petioles and rachises tissues of leaves fallen from infected trees were a good source for re-isolation of the pathogen; we proved that such petioles and rachises can effectively transfer the fungus to healthy Ailanthus seedlings via root infections. Host-specificity of the V. dahliae isolate VdGL16 was also determined on 40 non-target species/varieties/cultivars. The isolate caused disease in herbaceous species belonging to five botanical families: Asteraceae, Lamiaceae, Leguminoseae, Linaceae and Solanaceae. Given the difficulties in countering Ailanthus invasion with mechanical and chemical methods, the biological control using Verticillium may provide an efficient, low cost and sustainable control of this invasive species.