Nuclear rDNA Research Articles

Fasciola hepatica and Fasciola hybrid form co-existence in yak from Tibet of China: application of rDNA internal transcribed spacer.

Fasciolosis is a parasitic disease affecting humans and livestock, caused by digenean trematodes of the genus Fasciola, primarily F. hepatica and F. gigantica. This study investigates the coexistence of these species and their hybrids in yaks from Tibet, China. We analyzed the nuclear rDNA internal transcribed spacer (ITS) regions, including ITS1 and ITS2, through Sanger sequencing and Next-Generation Sequencing (NGS) to assess single-nucleotide polymorphisms (SNPs). Our results reveal that one specimen (NM008B) is identical to pure F. hepatica, while another (NM008A) contains genetic information from both F. hepatica and F. gigantica, indicating potential hybridization or introgression. The morphological analysis reveals that the collected adult F. hepatica specimens exhibit distinct characteristics, while the hybrid specimens display "intermediate" features of F. hepatica and F. gigantica. This study is the first to document the coexistence of F. hepatica and hybrid Fasciola forms in a single yak. The findings underscore the complexities of hybridization dynamics and the necessity for advanced molecular techniques in accurately identifying Fasciola species. Future research should focus on mitochondrial DNA and other nuclear gene analysis to further elucidate the nature of these hybrids and their ecological implications.

Integrative taxonomy reveals two new species of Maritrema Nicoll, 1907 (Digenea: Microphallidae Ward, 1901), parasitizing birds in the Gulf of Mexico.

Members of the genus Maritrema Nicoll, 1907 include endoparasites of aquatic birds and mammals, distributed worldwide. Adult specimens were collected from the intestines of three bird species (the great black hawks, Buteogallus urubitinga Gmelin; laughing gull, Leucophaeus atricilla Linnaeus; and the willet, Tringa semipalmata Gmelin) in three localities along the Gulf of Mexico. Photogenophores were sequenced for the large subunit (LSU) from nuclear rDNA, and the new sequences were aligned with other microphalloid sequences available in GenBank. The maximum likelihood and Bayesian inference analyses revealed three independent lineages, one corresponding to a previously described species, Maritrema corai Hernández-Orts, Pinacho-Pinacho, García-Varela & Kostadinova, 2016, and two representing two undescribed species. These two new species are described in the current study. Maritrema itzamnai n. sp. can be morphologically differentiated from its four congeneric species occurring in coastal areas of Mexico by having smaller oral (20-29 × 20-38 μm) and ventral (20-39 × 19-33 μm) suckers. In addition, Maritrema itzamnai n. sp. possess annular vitellarium instead of a horseshoe-shaped vitellarium. Maritrema kukulkanni n. sp. can be distinguished from its congeneric species reported from Mexico by possessing a larger oesophagus (44-117μm) and by having a vitellarium distributed in both the hindbody and forebody. Maritrema corai is the third species recorded in this study parasitizing the great black hawk (Buteogallus urubitinga); this represents a new host and locality record, expanding its distribution range from the Mexican Pacific Ocean to the Gulf of Mexico.

An integrative taxonomic and molecular identification of Calvatia holothurioides (Lycoperdaceae): the present status of genus Calvatia in India

Genus Calvatia belongs to a gasteroid fungi which are commonly known as a puffball mushrooms including around 47 species worldwide. Approximately 25 species of the genus Calvatia have been reported previously in India from various forest regions; however, only 15 species of these are accepted on their current taxonomic status because several of them are synonymized with other genera or species. The present study reports the distribution of Calvatia holothurioides which was collected from Jambughoda Wildlife Sanctuary in Gujarat state, India. The identification was carried out based on the morphological characteristics as well as molecular phylogenetic analysis using nuclear rDNA Internal Transcribed Spacer (ITS) and rDNA LSU gene sequences data. Nucleotide sequences of different gene loci (ITS and LSU) were submitted into the BOLD data system for DNA barcoding. Additionally, this study also provides a literature-based checklist for the present status of Calvatia species reported in India so far.

Morpho-phylogenetic evidence reveals novel species and new records of Nigrograna (Nigrogranaceae) associated with medicinal plants in Southwestern China.

During a survey of saprobic fungal niches in Southwestern China, eighteen ascomycetous collections of Nigrograna (Nigrogranaceae, Pleosporales, Dothideomycetes) were found on dead branches of medicinal plants. These taxa were characterized and identified based on morphological and culture characteristics, and phylogenetic analyses of a combined the internal transcribed spacer region of rDNA (ITS), nuclear large subunit rDNA (28S, LSU), RNA polymerase second-largest subunit (rpb2), nuclear small subunit rDNA (18S, SSU), and translation elongation factor 1-alpha (tef1-α) sequence dataset also confirmed their placement. As a result, four novel species, namely Nigrogranacamelliae, N.guttulata, N.longiorostiolata and N.neriicola were described. Additionally, four new host records of N.acericola, N.magnoliae, N.oleae and N.thymi were introduced. Furthermore, this study addresses the taxonomic status of N.trachycarpi, proposing its synonymy under N.oleae. Detailed illustrations, descriptions and informative notes for each newly identified taxon and novel host record are provided in this study.

A NEW SPECIES OF ACANTHOBOTHRIUM (CESTODA: ONCHOPROTEOCEPHALIDEA), PARASITE OF THE GIANT ELECTRIC RAY NARCINE ENTEMEDOR JORDAN AND STARKS, 1895 (BATOIDEA: TORPEDINIFORMES) FROM MEXICO.

Spiral valves from specimens of the giant electric ray Narcine entemedor Jordan & Starks, 1895 were examined in search of tapeworms at 2 localities of the Mexican tropical Pacific Ocean. Acanthobothrium oceguerai n. sp. is described herein based on material from Ventanilla, Oaxaca and from Acapulco Bay, Guerrero. The new species is a category 6 species, distinguished by being apolytic, retaining proglottids on the strobila until they are gravid, having strobila of 166-322 proglottids, having a small scolex and very long bothridia relative to the scolex, and having abaxial prongs that are short and thin in comparison to the axial prongs, which are longer and more robust. Acanthobothrium oceguerai n. sp. can be differentiated from other members of category 6 by the hooks, which are shorter, more robust, and smaller than those of the other members of this category. The phylogenetic analysis based on the 28S rRNA locus placed Acanthobothrium oceguerai n. sp. as sister to an unidentified species of larval Acanthobothrium from Philadelphia, Pennsylvania. In addition, sequences of the mitochondrial cytochrome oxidase subunit I gene and nuclear 18S rDNA were generated to provide support for future taxon sampling. Acanthobothrium oceguerai n. sp. is the fifth species of cestode reported from N. entemedor in the tropical eastern Pacific.

A new species of the Trichia botrytis complex from the cloud forest in Ecuador

The paper describes a new myxomycete species, Trichia tuberculata, from the Ecuadorian cloud forest. The phylogeny constructed with nuclear 18S rDNA and mitochondrial 17S rDNA sequences indicates that the taxon is closely related to the recently described species T. acetocorticola, T. nubila, and T. pinicola, all within the T. botrytis complex. The morphology of the inner peridium surface and capillitium also aligns T. tuberculata with these species. However, the distinctive tuberculate peridium, short elater tips, and perhaps the distribution range (so far found in the equatorial zone only) distinguish T. tuberculata from related taxa.

Freshwater ‘microcroissants’ shed light on a novel higher-level clade within the Trebouxiophyceae and reveal the genus Chlorolobion to be a trebouxiophyte

ABSTRACT The Trebouxiophyceae is a widespread and species-rich green algal class encompassing mostly coccoid algae with a simple spherical, ovoid or ellipsoidal outline. However, some poorly sampled lineages have evolved more elaborate shapes or even complex thalli, adding to the morphological diversity of the class. By investigating new and previously established strains, this study expands the range of morphologies exhibited by the class members by uncovering a clade of croissant-like trebouxiophytes. Phylogenetic analyses inferred from nuclear 18S rDNA and chloroplast rbcL sequences confirmed the monophyly of the ‘microcroissant’ clade, which we propose to be classified as a new family, Ragelichloridaceae. This family includes two novel genera, Ragelichloris and Navichloris, and the previously described Thorsmoerkia. The position of Ragelichloridaceae within Trebouxiophyceae stayed unresolved but chloroplast phylogenomics showed that the family belongs to the broader incertae sedis group that also includes Xylochloris and Leptosira. In addition, our study showed that the similar morphotype-bearing genus Chlorolobion, previously classified within Chlorophyceae, is a genuine trebouxiophyte, potentially related to Ragelichloridaceae.

Assessment of the Genetic Diversity of the Monogenean Gill Parasite Lamellodiscus echeneis (Monogenea) Infecting Wild and Cage-Reared Populations of Sparus aurata (Teleostei) from the Mediterranean Sea.

The diplectanid monogenean Lamellodiscus echeneis (Wagener, 1857) is a specific and common gill parasite of the gilthead seabream Sparus aurata Linnaeus, 1758, in the Mediterranean Sea. Few isolated molecular studies of this monogenean have been conducted, and its population structure and genetic diversity are poorly understood. This study represents the first analysis of the population genetics of L. echeneis, isolated from wild and cage-reared gilthead seabream from fifteen localities in both the Southern (Tunisia) and Northern (Italy and Spain) regions of the Mediterranean Sea, using nuclear ITS rDNA markers and a partial fragment of the mitochondrial gene cytochrome oxidase subunit I (COI). The phylogenetic trees based on the newly obtained dataset and the previously published sequences of L. echeneis corroborated the spread of only a single species throughout the Mediterranean Sea. The star-like haplotypes network, inferred by COI sequences, suggested a recent population expansion of L. echeneis. This is supported by the observed high haplotype diversity (Hd = 0.918) and low nucleotide diversity (Pi = 0.01595). Population structure-based AMOVA for two groups (the Adriatic Sea and the rest of the Mediterranean Sea) attributed 35.39% of the total variation to differences within populations, 16.63% to differences among populations within groups, and 47.99% to differences among groups. Fixation indices were significant, with a high FST value (0.64612), likely related to the divergence of the parasite populations from the Adriatic Sea and other Mediterranean regions. Phylogenetic analyses grouped all samples into the main clade corresponding to L. echeneis from several localities. This study provides insight into the genetic variation between L. echeneis populations, and did not show a clear genetic structure between populations of L. echeneis throughout Tunisian, Italian, and Spanish localities, which can be attributed to the considerable gene flow between the populations favoured by the potential for host dispersion within the Mediterranean Sea. Finally, haplotypes shared between wild and cage-reared hosts provided evidence for the potential for cross-infection between wild and farmed hosts in the Mediterranean Sea.

Detection of Podosphaera macularis in Air Samples by Quantitative PCR.

Detection and quantification of pathogen propagules in the air or other environmental samples is facilitated by culture-independent assays. We developed a quantitative PCR assay for the hop powdery mildew fungus, Podosphaera macularis, for detection of the organism from air samples. The assay uses primers and a TaqMan probe designed to target species-specific sequences in the 28S large subunit of the nuclear ribosomal DNA. Analytical sensitivity was not affected by the presence of an exogenous internal control or potential PCR inhibitors associated with DNA extracted from soil. The level of quantification of the assay was between 200 and 350 conidia when DNA was extracted from a fixed number of conidia. The assay amplified all isolates of P. macularis tested and had minimal cross-reactivity with other Podosphaera species when assayed with biologically relevant quantities of DNA. Standard curves generated independently in two other laboratories indicated that assay sensitivity was qualitatively similar and reproducible. All laboratories successfully detected eight unknown isolates of P. macularis and correctly discriminated Pseudoperonospora humuli and a water control. The usefulness of the assay for air sampling for late-season inoculum of P. macularis was demonstrated in field studies in 2019 and 2020. In both years, airborne populations of P. macularis in hop yards were detected consistently and increased during bloom and cone development.

Morpho-molecular characterization of phoma-like fungi from Morus alba in northern Thailand; a novel species (Boeremia albae) and a new host record (B. maritima)

Boeremia was established to accommodate phoma-resembling fungi. Its species occur in terrestrial ecosystems as endophytes, saprobes and pathogens, except one species reported from a marine ecosystem. Boeremia species are characterized by hyaline, thin-walled, and aseptate (occasionally 1(–2)-septate) conidia that are variable in shape, and hyaline, straight or slightly curved, thick-walled, and 1-septate ascospores that are usually constricted at the septum. In the past, host associations were used to delimit Boeremia species. However, since Boeremia taxa have overlapping morphological characters and are cryptic, it renders taxonomic identification arduous. Therefore, the use of other approaches including multi-gene phylogenetic analyses are imperative. Recommended DNA markers for species delineation are the internal transcribed spacer (ITS, nuclear rDNA consisting of ITS1-5.8S-ITS2) and large subunit (28S, D1–D2 domains of nuclear 28S rDNA) loci, and the genes for actin (ACT1), beta-tubulin (TBB1), RNA polymerase 2 (RPB2) and translation elongation factor 1α (TEF1). Here, we applied morphological and molecular phylogenetic analyses to establish a new taxon (B. albae), and a new host and geographical record for B. maritima associated with leaf spots of Morus alba (Moraceae) in northern Thailand. By providing sequence data for three additional gene regions, our phylogenetic analyses impart a stable phylogenetic placement of the ex-type strain of B. maritima, as illustrated. This is the first study that reports Boeremia species from M. alba, and B. maritima from a terrestrial habitat.

DESCRIPTIONS AND PHYLOGENETIC AFFINITIES OF TWO NEW SPECIES OF RHABDIAS STILES AND HASSALL, 1905 (NEMATODA: RHABDIASIDAE) FROM NORTH AMERICAN FROGS: ARE WE STILL ONLY SCRATCHING THE SURFACE?

Two new species of lung-dwelling nematodes are described from North American frogs: Rhabdias aurorae n. sp. from Rana aurora and Rhabdias conni n. sp. from Rana clamitans and Rana catesbeiana from Arkansas; the latter species was also found in Oklahoma and Georgia. Rhabdias aurorae n. sp. differs from other Nearctic congeners in the combination of the following characteristics: buccal capsule 22-25 μm wide, elongated tail covered with inflated cuticle, esophagus with prominent dilatation in anterior part and 6 small circumoral lips. Rhabdias conni n. sp. is morphologically closest to Rhabdias ranae Walton, 1929 and Rhabdias joaquinensisIngles, 1936; it differs from them in the shape of lateral pseudolabia, the dimensions of the body, and the egg size. Both new species were found to be significantly different from the Nearctic congeners in the nucleotide sequences of nuclear ribosomal DNA (18S-ITS-28S region), 12S, and CO1 mitochondrial genes. The 2 new species differ from other currently sequenced Nearctic congeners by 1.1-2.7% of nucleotide positions in the nuclear rDNA region, 1.3-3.4% in the 12S gene, and 3.4-9.4% in CO1 gene. Molecular phylogenetic analysis based on nuclear ribosomal DNA sequences placed both new species into the clade consisting of Nearctic and Neotropical Rhabdias spp. The position of Rh. aurorae n. sp. within the clade is uncertain because of a polytomy, but Rh. conni n. sp. is nested within the "Rh. joaquinensis complex" related to Rh. ranae and Rhabdias tarichaeKuzmin, Tkach, and Snyder, 2003. The phylogenetic analysis based on nuclear ribosomal DNA sequences has revealed 3 evolutionary host-switching events from anuran to caudatan hosts among Rhabdias spp. that occurred in the Nearctic and Palearctic. The molecular phylogeny also suggests that Rhabdias may have originally evolved in what is now Africa.

Morphological and phylogenetic analyses reveal four species of myxomycetes new to China.

Four species of myxomycetes (Arcyria pseudodenudata, Diderma europaeum, Lycogala irregulare, and Trichia armillata) new to China were observed via light microscope and scanning electron microscope, and detailed descriptions and illustrations are provided, along with comparisons with related species. Among them, A. pseudodenudata was discovered for the first time outside of the type locality, D. europaeum was discovered for the first time outside of Europe, and L. irregulare and T. armillata were reported again after being named. Phylogenetic analyses based on nuclear 18S rDNA and elongation factor-1 alpha sequences or nuclear 18S rDNA and cytochrome oxidase subunit I sequences was performed to provide a molecular basis for morphological identification. These specimens were deposited in the Herbarium of Fungi of Nanjing Normal University.

Cryptic invasions of Pseudopolydora (Annelida: Spionidae), with description of a new species from Queensland, Australia

During surveys worldwide, we collected adult and larval specimens of Pseudopolydora Czerniavsky, 1881 similar to P. achaeta Radashevsky & Hsieh, 2000 and P. rosebelae Radashevsky & Migotto, 2009 far from their type localities in Taiwan and Brazil, respectively. Analysis of sequence data of five gene fragments: mitochondrial COI and 16S rDNA, nuclear 18S rDNA and 28S rDNA, and Histone 3 (3114 bp in total) of individuals from Nha Trang Bay (Vietnam), São Paulo (Brazil), Florida (USA), and the Caribbean Sea (Martinique) confirmed their conspecificity with P. achaeta from Taiwan. Based on the morphology, we also report this species for the first time for the East Sea (South Korea) and provide new records for the north-western part of the Sea of Japan (Russia). The analysis also showed the conspecificity of worms from Vietnam with P. rosebelae from Brazil. Based on the morphology, we also report P. rosebelae for the first time for the Caribbean Sea (Belize), South China Sea (Thailand), and Hawaii. Developed pelagic larvae of P. achaeta and P. rosebelae from Nha Trang Bay are described and illustrated. A worm from the Great Barrier Reef (Queensland, Australia) earlier identified as P. cf. rosebelae, is referred to the new species Pseudopolydora nivea sp. nov. Following our earlier hypothesis about the origin of Pseudopolydora in the Indo-West Pacific Ocean, we explain the occurrence of these worms, including Pseudopolydora floridensis Delgado-Blas, 2008, on the Atlantic coasts of North and South America by unintentional transportation in ballast water, followed by successful invasions. We assume that the penetration of warm waters of the East China Sea through the Korea Strait and the warming of the waters of the Sea of Japan have led to the extension of the range of tropical-subtropical P. achaeta to the north-western part of the Sea of Japan.

Expansions and contractions of repetitive DNA elements reveal contrasting evolutionary responses to the polyploid genome shock hypothesis in Brachypodium model grasses.

Brachypodium grass species have been selected as model plants for functional genomics of grass crops, and to elucidate the origins of allopolyploidy and perenniality in monocots, due to their small genome sizes and feasibility of cultivation. However, genome sizes differ greatly between diploid or polyploid Brachypodium lineages. We have used genome skimming sequencing data to uncover the composition, abundance, and phylogenetic value of repetitive elements in 44 representatives of the major Brachypodium lineages and cytotypes. We also aimed to test the possible mechanisms and consequences of the "polyploid genome shock hypothesis" (PGSH) under three different evolutionary scenarios of variation in repeats and genome sizes of Brachypodium allopolyploids. Our data indicated that the proportion of the genome covered by the repeatome in the Brachypodium species showed a 3.3-fold difference between the highest content of B. mexicanum-4x (67.97%) and the lowest of B. stacei-2x (20.77%), and that changes in the sizes of their genomes were a consequence of gains or losses in their repeat elements. LTR-Retand and Tekay retrotransposons were the most frequent repeat elements in the Brachypodium genomes, while Ogre retrotransposons were found exclusively in B. mexicanum. The repeatome phylogenetic network showed a high topological congruence with plastome and nuclear rDNA and transcriptome trees, differentiating the ancestral outcore lineages from the recently evolved core-perennial lineages. The 5S rDNA graph topologies had a strong match with the ploidy levels and nature of the subgenomes of the Brachypodium polyploids. The core-perennial B. sylvaticum presents a large repeatome and characteristics of a potential post-polyploid diploidized origin. Our study evidenced that expansions and contractions in the repeatome were responsible for the three contrasting responses to the PGSH. The exacerbated genome expansion of the ancestral allotetraploid B. mexicanum was a consequence of chromosome-wide proliferation of TEs and not of WGD, the additive repeatome pattern of young allotetraploid B. hybridum of stabilized post-WGD genome evolution, and the genomecontraction of recent core-perennials polyploids (B. pinnatum, B. phoenicoides) of repeat losses through recombination of these highly hybridizing lineages. Our analyses have contributed to unraveling the evolution of the repeatome and the genome size variation in model Brachypodium grasses.

Taxonomy and fatty acid profile of Picochlorum geojense sp. nov. (Chlorophyta, Trebouxiophyceae) from Korean coastal waters

ABSTRACT The microalgal genus known as Picochlorum, belonging to the green algae family Trebouxiophyceae, is recognized for its rapid growth and high lipid productivity. A Picochlorum strain (GJ210610) was recently established from the waters off Geoje, South Sea of Korea. Picochlorum sp. GJ210610 was a single, green ellipsoidal cell with an average size ranging from 1 to 2 µm. Its reproduction occurred through autosporulation. A phylogenetic tree based on nuclear small-subunit rDNA sequences revealed that this strain formed a unique branch, distinct from other Picochlorum species. Furthermore, the secondary structure of the ITS2 sequences was revealed for the first time within Picochlorum. The newly discovered Picochlorum sp. grew between 15°C and 25°C, with survival ceasing at 30°C. The maximum specific growth rate was measured at 0.91 d−1 at 25°C. The fatty acids of Picochlorum sp. GJ210610 were palmitic acid, oleic acid, linoleic acid, and alpha-linolenic acid. Based on morphological and phylogenetic analyses, we propose Picochlorum geojense sp. nov. (GJ210610) as a new species. The results of the present study may enhance our knowledge of the taxonomy and ecological characteristics of Picochlorum.

A new omphalinoid species, Entoloma khushabense (Entolomataceae, Basidiomycota) from Punjab, Pakistan

A new species, Entoloma khushabense, is described and illustrated here based on morphological and molecular evidence from Pakistan. The key features of the new species that make it distinct from closely related taxa include the presence of conspicuous cheilocystidia and caulocystidia, and hyphae having constrictions in the walls. Morphological data and phylogenetic analyses based on internal transcribed spacer (ITS) and large sub-unit (LSU) regions of nuclear rDNA support that the present species differs from previously known members of Entoloma.

Taxonomy and phylogeny of three species in Perichaena sensu lato (Myxomycetes = Myxogastria) from China

After Gulielmina was erected and Ophiotheca was resurrected based on some species originally included in Perichaena (Trichiaceae, Trichiales, Myxomyxetes), some specimens from the Herbarium of Fungi of Nanjing Normal University previously identified as Perichaena species were reexamined from morphological and two-gene (nuclear 18S rDNA and elongation factor-1 alpha) phylogenetic perspectives. In this study, two new myxomycete species, Gulielmina subreticulospora and Ophiotheca dictyospora, are described. Gulielmina subreticulospora shows the following character combination: branched plasmodiocarps, single peridium with circular protrusions in the inner surface, capillitium (2.4–)2.8–3.0(–3.4) μm in diameter, spores (7.4–)8.0–8.5(–9.0) μm in diameter and sub-reticulated. Ophiotheca dictyospora shows the following character combination: sessile sporocarps to short plasmodiocarps, single peridium with a densely irregular network and protrusions in the inner surface, capillitium (2.7–)3.5–5.0(–7.1) μm in diameter, uneven, decorated with spines of uneven size, spores (7.7–)8.2–8.6(–9.4) μm in diameter including obviously complete cristate reticulation with serrated edges, with deep and clear grids. Both new taxa were compared with related species and their genetic isolation was statistically tested. Moreover, a comprehensive morphological description and a detailed figure plate are provided for Perichaena verrucifera, and its phylogenetic position is determined.

Uncovering further diversity of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) in South American bats

Ochoterenatrema Caballero, 1943 is a genus of lecithodendriid digeneans that prior to this study included 8 species parasitic in bats in the Western Hemisphere. Species of Ochoterenatrema possess a unique morphological feature in form of the pseudogonotyl on the sinistral side of the ventral sucker. In this study, we describe 2 new species of Ochoterenatrema from bats in Ecuador. The new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles, length of oesophagus, sucker ratio and the body shape, among other features. We have generated partial nuclear 28S rDNA and mitochondrial cox1 gene DNA sequences from both new species. The newly obtained sequences were used to differentiate among species and study the phylogenetic interrelationships among Ochoterenatrema spp. The internal topology of the clade was weakly supported, although the cox1 tree was much better resolved than the 28S tree. Comparison of sequences revealed 0–1.2% interspecific divergence in 28S and 3.3–20.5% interspecific divergence in cox1 among Ochoterenatrema spp. The new findings demonstrate that bats in South America likely harbor multiple additional undescribed species of Ochoterenatrema. More extensive sampling from broader geographic and host ranges, especially in North America, should allow for a better understanding of the evolution of host associations and morphological traits of this lineage of lecithodendriid digeneans.

Characterization of Alternaria blotch disease of apple in Himachal Pradesh, India: insights on morphology, pathogenicity, and molecular features.

Alternaria blotch disease in Himachal Pradesh, India, caused by Alternaria spp., adversely affects apple cultivars, resulting in reduced fruit size and quality accompanied by premature leaf fall. Sixteen Alternaria isolates from apple growing regions underwent comprehensive analysis including morphology, pathogenicity, and molecular characterization. Variations in conidiophore and conidia dimensions, shapes, and divisions were observed among isolates. Pathogenicity assays revealed differences in incubation periods, latent phases, and disease responses. Molecular characterization via nuclear ITS rDNA and RAPD analysis indicated 99-100% homology with Alternaria alternata, Alternaria mali, and other Alternaria spp., with a close phylogenetic relationship to Chinese isolates. Differentiation of isolates based on origin, cultural characteristics, and morphology was achieved using RAPD markers. The study identifies diverse genotypes and morphotypes of Alternaria contributing to apple blotch disease in Himachal Pradesh. These findings highlight the complexity of the pathogenic environment and hold significant implications for disease management in apple orchards.

Expanding on expansus: a new species of Scaphanocephalus from North America and the Caribbean based on molecular and morphological data.

Members of the genus Scaphanocephalus mature in accipitrids, particularly osprey, Pandion haliaetus, with metacercaria causing Black Spot Syndrome in reef fishes. In most of the world, only the type species, Scaphanocephalus expansus (Creplin, 1842) has been reported. Recent molecular studies in the Western Atlantic, Mediterranean and Persian Gulf reveal multiple species of Scaphanocephalus, but have relied on 28S rDNA, mainly from metacercariae, which limits both morphological identification and resolution of closely related species. Here we combine nuclear rDNA with mitochondrial sequences from adult worms collected in osprey across North America and the Caribbean to describe species and elucidate life cycles in Scaphanocephalus. A new species described herein can be distinguished from S. expansus based on overall body shape and size. Phylogenetic analysis of the whole mitochondrial genome of Scaphanocephalus indicates a close relationship with Cryptocotyle. We conclude that at least 3 species of Scaphanocephalus are present in the Americas and 2 others are in the Old World. Specimens in the Americas have similar or identical 28S to those in the Mediterranean and Persian Gulf, but amphi-Atlantic species are unlikely in light of divergence in cytochrome c oxidase I and the lack of amphi-Atlantic avian and fish hosts. Our results provide insight into the geographic distribution and taxonomy of a little-studied trematode recently linked to an emerging pathology in ecologically important reef fishes.