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Genetic sub-populations (clades) of cosmopolitan marine diatom Pseudo-nitzschia pungens might have distinct habitats, and their hybrid zone is suspected in higher latitude area of the West Pacific area, however, it is still unrevealed because of technical difficulties and lack of evidences in natural environments. The aim of this study is to investigate the habitat characteristics of each clade of P. pungens on geographical distribution with the habitat temperature ranges of each clade and to reveal their hybrid zone in the West Pacific area. We employed the 137 number of nucleotide sequences of P. pungens and its sampling data (spatial and temporal scale) originated from the West Pacific area, and used field application of qPCR assay for intra-specific level of P. pungens . Only two genotypes, clade I and III, were identified in the West Pacific area. Clade I was distributed from 39 to 32.3°N, and clade III were from 1.4 to 34.4°N. The estimated habitat temperature for the clade I and clade III ranges were 8.1–26.9 °C and 24.2–31.2 °C, respectively. The latitudinal distributions and temperature ranges of each clade were significantly different. The qPCR assay employed, and results suggested that the hybrid zone for clade I and III has been observed in the southern Korean coasts, and clade III might be introduced from the Southern Pacific area. The cell abundances of clade III were strongly related with the higher seawater temperature and warm current force. This study has defined distinct habitat characteristics of genetically different sub-populations of P. pungens , and revealed its hybrid zone in natural environment for the first time. We also provided strong evidences about dispersion of the population of clade III to higher latitude in the West Pacific area. [ABSTRACT FROM AUTHOR]

Three genetic sub-populations (clade I, II and III) of Pseudo-nitzschia pungens , the potential toxic marine diatom, are known to have distinguishable growth characteristics under different culture conditions and distinct distributed patterns in the world. However, to date their exact eco-physiological traits are unrevealed in fields due to lack of the method to detect and/or measure abundances of each sub-populations, hence, the qPCR (quantitative real-time polymerase chain reaction) assay was developed to detect and quantify the P. pungens cells of each clade. Designed two specific primer sets, Pcla12F/R (for clade I and II) and Pcla3F/R (for clade III) only could amplify each target genomic DNA. The, significant linear relationships (R 2 > 0.998) was established between C t (threshold cycle) value and the log of cell abundance for each clade. Through the melting curve analysis, comparisons for gene copy numbers among the three clades and spike test for field study, our qPCR assay was reliable to quantify the cell numbers of each clade. There was strong linear correlation (R 2 > 0.990) between cell abundances as estimated by qPCR assay and direct counting via light microscope in spike test, and 0.24 (clade I), 0.25 (clade II) and 0.33 (clade III) P. pungens cells per mL were detected markedly upon the use of specific two-primer set. Finally, developed qPCR assay was applied on field samples successfully. Our study implicate that our qPCR assay is an accurate and sensitive technique to estimate the cell abundances of each clade of P. pungens in field works. [ABSTRACT FROM AUTHOR]

• Prorocentrum donghaiense is the most notorious HAB species in the East China Sea. • Blooms of P. donghaiense in the East China Sea are results of temperature increase and eutrophication. • Phosphorous deficiency plays a critical role in regulating P. donghaiense blooms. • The major harmful consequence of P. donghaiense blooms is disruption of ecosystems. Prorocentrum donghaiense Lu (also identified as Prorocentrum shikokuense Hada and Prorocentrum obtusidens Schiller) is a bloom-forming dinoflagellate species distributed worldwide. Blooms of P. donghaiense occur annually in adjacent waters of the East China Sea (ECS), especially in the waters near the Changjiang River Estuary. Blooms of this species have also been reported in nearby Japanese and Korean waters. There has been an apparent bloom-forming species succession pattern in the ECS since 2000, with diatom blooms in the early spring, shifting to long-lasting and large-scale dinoflagellate blooms dominated by P. donghaiense during the spring, and finally ended by diatom and/or Noctiluca scintillans blooms in summer. These bloom succession patterns were closely correlated with changes in environmental factors, such as temperature increase and anthropogenic eutrophication. Decreasing silicate by the construction of the Three Gorges Dam and increasing dissolved inorganic nitrogen flux were mainly influenced by high intensity human activities in the Changjiang River watershed, resulting in low Si/N ratio and high N/P ratios, possibly accelerating outbreak of P. donghaiense blooms. Phosphorous deficiency might be the most critical factor controlling the succession of microalgal blooms from diatoms to dinoflagellates. Prorocentrum donghaiense is a nontoxic species, but it can disrupt marine ecosystem by decreasing phytoplankton biodiversity and changing the structure of the food chain. Prorocentrum donghaiense blooms in the ECS have been intensively studied during the last two decades. Several possible mechanisms that contribute or trigger the annual blooms of this species have been proposed, but further research is required particularly on the aspect of nutrient budget, ecosystem impacts, as well as social-economic impact assessment. [ABSTRACT FROM AUTHOR]

• Aside from East Asia, historical records also showed the wide distribution of harmful raphidophyte Chattonella in Southeast Asian coastal waters. • Only one genotype, Chattonella marina complex, has been observed in East Asia. • Two Chattonella genotypes, C. marina complex and C. subsalsa , have occurred in Southeast Asia and were associated with fisheries damage. • Oboe-shaped mucocyst was observed from the cultures of C. marina from Indonesia, Malaysia, Japan and Russia for the first time. Red tides and associated fisheries damage caused by the harmful raphidophyte Chattonella were reassessed based on the documented local records for 50 years to understand the distribution and economic impacts of the harmful species in the Western Pacific. Blooms of Chattonella with fisheries damage have been recorded in East Asia since 1969, whereas they have been only recorded in Southeast Asia since the 1980s. Occurrences of Chattonella have been documented from six Southeast Asian countries, Indonesia, Malaysia, Philippines, Singapore, Thailand and Viet Nam, with mass mortalities mainly of farmed shrimp in 1980–1990s, and farmed fish in 2000–2010s. These occurrences have been reported with the names of C. antiqua, C. marina, C. ovata, C. subsalsa and Chattonella sp., owing to the difficulty of microscopic species identification, and many were not supported with molecular data. To determine the distribution of C. marina complex and C. subsalsa in Southeast Asia, molecular phylogeny and microscopic observation were also carried out for cultures obtained from Indonesia, Malaysia, Japan, Philippines, Russia, Singapore and Thailand. The results revealed that only the genotype of C. marina complex has been detected from East Asia (China, Japan, Korea and Russia), whereas both C. marina complex (Indonesia and Malaysia) and C. subsalsa (Philippines, Singapore and Thailand) were found in Southeast Asia. Ejection of mucocysts has been recognized as a diagnostic character of C. subsalsa , but it was also observed in our cultures of C. marina isolated from Indonesia, Malaysia, Japan, and Russia. Meanwhile, the co-occurrences of the two harmful Chattonella species in Southeast Asia, which are difficult to distinguish solely based on their morphology, suggest the importance of molecular identification of Chattonella genotypes for further understanding of their distribution and negative impacts. [ABSTRACT FROM AUTHOR]