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High-resolution DNA metabarcoding of modern surface sediments uncovers a diverse assemblage of dinoflagellate cysts in the Pacific and Arctic Oceans
Resting cysts of dinoflagellates can persist in sediments, seeding harmful algal blooms (HABs). A DNA metabarcoding approach was employed, targeting the large subunit ribosomal (LSU D1–D2) and the internal transcribed spacer (ITS1) to investigate the diversity and biogeography of dinoflagellate cysts from the South China Sea to the Chukchi Sea. The LSU and ITS1 datasets identified 196 and 118 species, respectively, with only 59 dinoflagellate cyst species revealed by both approaches. Eleven cyst species of potentially toxic dinoflagellates and 82 species previously unknown as cyst producers were detected. Cysts of Heterocapsa cf. horiguchii, Heterocapsa minima, Heterocapsa iwatakii, Heterocapsa rotundata, and Heterocapsa steinii were documented through germination for the first time, with the latter three species also detected via metabarcoding. This study provides critical insights into the diversity and biogeography of dinoflagellate cysts by highlighting the complementary detection capabilities of LSU and ITS1 molecular markers and their trans-latitudinal distribution patterns. The identification of potentially toxic cysts and their ecological distributions offers crucial information on the ecology of harmful dinoflagellates. These findings underscore the importance of molecular techniques in monitoring dinoflagellate cysts.
Integrative characterization of Ostreopsis cf. ovata (Dinophyceae) in the central and southwestern Gulf of California: Morphology, molecular phylogeny, and toxicity assessment
Potentially toxic dinoflagellates of the genus Ostreopsis are expanding their distribution globally, with a marked increase in the frequency of Benthic Harmful Algal Blooms (BHABs). Six of the twelve species of Ostreopsis produce a highly potent non-proteinaceous toxin known as palytoxin (PLTX) and its analogs, posing a significant threat to human and environmental health. The study of BHAB species, particularly the genus Ostreopsis, is a recent development in Mexico. With a comprehensive approach, this study aimed to taxonomically and toxicologically characterize Ostreopsis strains from three locations in the central and southeastern Gulf of California. Morphometric and tabular analyses suggested that Ostreopsis cf. ovata and O. fattorussoi are the most probable morphospecies. However, phylogenetic analyses inferred from LSU rDNA (D1-D3) and ITS rDNA sequences further narrowed our results to the Ostreopsis ovata species complex, which also suggests its dominance in the region. In a composite sample (three strains from each of the three locations) chemically analyzed with LC-MS/MS, no intact palytoxins were found; however, amine aldehyde fragment derivatives common to PLTX, OST-D, OVTX, 42-OH PLTX, deoxy PLTX, and isobaric PLTX, were found at low intracellular concentrations. These findings were consistent with an Artemia franciscana bioassay, as the mortality rate after 48 h of exposure was below 10 % and showed no significant differences (p > 0.05) between the control and experimental conditions. Therefore, it is concluded that the tested strains of O. cf. ovata do not currently represent a risk to human or ecosystem health.
Morpho-molecular diversity and host range dynamics of squash powdery mildew pathogens prevailing in Pothwar Plateau, Pakistan
Squash (Cucurbita spp.) is an economically important vegetable crop in Pakistan, yet its productivity is severely impacted by powdery mildew. Despite frequent outbreaks, limited data exist on its causal agents and host range in Pakistan. This study aimed to assess the prevalence, incidence, characterization and host range of associated squash powdery mildew (SPM) pathogens. Surveys conducted during 2020 and 2021 in the Pothwar region revealed 100% disease prevalence with mean incidence of 50.47% and 59.78%, respectively. Disease incidence was positively correlated with humidity and rainfall. Healthy plant assays for culture maintenance and preservation of inoculum at −20 °C were found the most efficient for maintaining the pathogen virulence even after a year whereas, 500 mg powder/L (3.4 × 104 spores) was standardized as optimum inoculum load for the onset of utmost disease severity. Microscopic observation of each sample identified two coexisting SPM causal agents i.e. Podosphaera xanthii (Px.) and Golovinomyces cichoracearum (Gc.). Nucleotide sequences of highly virulent isolates from pathogenicity tests confirmed the species identity and phylogenetic analyses revealed genetic similarity with global isolates. All cucurbit crops were susceptible to SPM whereas, the weeds were infected by other powdery mildew species. Interestingly, sunflower (both wild and cultivated) (Helianthus annuus) was found infected with both SPM species which confirms the sunflower as potential alternate host for SPM.

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