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Heterocapsa bohaiensis sp. nov. (Peridiniales: Dinophyceae): a novel marine dinoflagellate from the Liaodong Bay of Bohai Sea, China
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Jie XIAO1, 2, †, Na SUN3, †, Yiwen ZHANG4, Ping SUN1, Yan LI1, 2, Min PANG1, Ruixiang LI1, *
Acta Oceanologica Sinica | 2018, 37(10) : 18 - 27
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Acta Oceanologica Sinica | 2018, 37(10): 18-27
• Articles •
Heterocapsa bohaiensis sp. nov. (Peridiniales: Dinophyceae): a novel marine dinoflagellate from the Liaodong Bay of Bohai Sea, China
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Jie XIAO1, 2, †, Na SUN3, †, Yiwen ZHANG4, Ping SUN1, Yan LI1, 2, Min PANG1, Ruixiang LI1, *
Affiliations
  • 1 Key Laboratory of Science and Engineering for Marine Ecology and Environment, the First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China
  • 2 Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China
  • 3 Guanghe Crab Industry Limited Company, Panjin 124200, China
  • 4 School of Food and Environment, Dalian University of Technology, Panjin 124221, China
Published: 2018-10-25 doi: 10.1007/s13131-018-1296-z
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A small armed dinoflagellate bloomed in the aquaculture ponds off the coast of Liaodong Bay, Bohai Sea of China, resulting in heavy mortalities of the cultured prawns (Penaeus japonicus) and larvae of Chinese mitten handed crabs (Eriocheir sinensis). The bloom-forming species was successfully isolated, and cellular morphology of the specimen was consequently investigated through light, fluorescent and electron microscopy. The small ((14.4±1.6) μm in length) ellipsoid cells show typical Heterocapsa thecal plate arrangement (Po, cp, 5′, 3a, 7′′, 6c, 5s, 5′′′, 2′′′′). The episome is evidently bigger than the hyposome. One to three spherical pyrenoids are located above or beside the large elongated nucleus. The body scale is characterized by a triangle basal plate with one central upright and nine peripheral spines. Above all, Heterocapsa bohaiensis could be distinguished from other Heterocapsa species by the combination of the cell size, morphology, cellular structure and body scale. Sequence analyses of both ITS and LSU regions reveal the significant genetic divergence between H. bohaiensis and other established species in this genus, further supporting novelty of this species. Noticeably, different sample treatment methods resulted in morphological variation of the apical pore complex (APC) of H. bohaiensis, which needs to be taken into account in future study.

Heterocapsa  /  harmful algal bloom  /  thecal plate  /  body scale  /  ITS
Jie XIAO, Na SUN, Yiwen ZHANG, Ping SUN, Yan LI, Min PANG, Ruixiang LI. Heterocapsa bohaiensis sp. nov. (Peridiniales: Dinophyceae): a novel marine dinoflagellate from the Liaodong Bay of Bohai Sea, China[J]. Acta Oceanologica Sinica, 2018 , 37 (10) : 18 -27 . DOI: 10.1007/s13131-018-1296-z
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1 Introduction
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The genus Heterocapsa was originally established by Stein in the 1880s based on the type species of Heterocapsa triquetra (=Glenodinium triquetrum Ehrenberg). A number of species were subsequently transferred into this genus, including H. pacifica, H. niei, H. illdefina and H. rotundata (Loeblich III, 1968; Herman and Sweeney, 1976; Hansen, 1995). Since the 1990s, more species were discovered and described, along with H. circularisquama, an HAB species blooming off the west coasts of Japan (Horiguchi, 1995, 1997; Matsuyama et al., 2001; Iwataki et al., 2002a, 2003, 2004, 2009; Tamura et al., 2005). So far, the genus Heterocapsa comprises about 19 species, characterized with a general plate formula of Po, cp, 5′, 3a, 7′′, 6c, 5–8 s, 5′′′, 0- lp, 2′′′′ (Loeblich III et al., 1981; Morrill and Loeblich III, 1981; Hansen, 1995; Horiguchi, 1995; Iwataki et al., 2002a, 2004; Gómez, 2005, 2012), an elongate anterior sulcal plate and presence of minute three-dimensional body scales (Pennick and Clarke, 1977; Morrill and Loeblich III, 1981, 1983; Iwataki et al., 2004). Although the taxonomic relationship between Heterocapsa and its closely-related genera (e.g., Cachonina) was still unresolved (Morrill and Loeblich III, 1981; Horiguchi, 1995), species within Heterocapsa could be distinguished by cell morphology, relative position and shape of the nucleus and pyrenoids, and the fine structure of the body scales (Iwataki et al., 2004; Iwataki, 2008).
Heterocapsa spp. are distributed widely in the coastal regions around the world, and some bloomed densely causing detrimental ecological impacts in some regions. Heterocapsa circularisquama was originally identified in Japanese coast, and intensive blooms of this species occurred in western Japan since 1988, resulting in catastrophic mortalities of the farmed molluscan shellfishes (Matsuyama et al., 1995, 2001). Later, Iwataki et al. (2002b) reported the similar H. circularisquama cells in the fixed backup samples of red tides occurred in Hong Kong in the 1980s, speculating a wider distribution of this species along the western Pacific. Although systematic research on toxicity and harmful effects of H. circularisquama on various organisms are still ongoing (Salcedo et al., 2012; Basti et al., 2015, 2016; Nishiguchi et al., 2016), a photosensitizing hemolytic toxin, presumably a novel porphyrin derivative, has been successfully isolated and characterized from H. circularisquama (Sato et al., 2002; Miyazaki et al., 2005). Heterocapsa circularisquama is the only toxic species reported in this genus so far. Other species, H. triquetra as an example, may form dense blooms in the coastal and estuary waters, while no evident harmful impacts have been reported (Kim, 1997; Lindholm and Nummelin, 1999; Kononen et al., 1999).
Except the H. circularisquama blooms in Hong Kong (Iwataki et al., 2002b), little was known about Heterocapsa spp. along the coasts of China. Two Heterocapsa species (H. minima and H. triquetra) were listed in the historical records as the common dinoflagellates in the East China Sea (ECS; Liu, 2008), while no descriptions on morphology and distribution were provided. Blooms of H. circularisquama and H. rotundata were also reported sporadically in the Changjiang (Yangzi River) Estuary of ECS, coasts of Dalian and Qingdao in recent years (Du, 2005; Wang et al., 2006; Liu et al., 2014). Whereas, the details on morphology and physiology of the suspicious cells were not reported, resulting in doubtful identity of these blooming cells (Liu et al., 2009; Zheng, 2009). In 2008 and 2012, dense-blooming of a small dinoflagellate occurred in the aquaculture ponds in Panjin, Liaodong Bay of Bohai Sea, causing heavy mortalities of the cultured prawns (Penaeus japonicus) and larvae of Chinese mitten handed crabs (Eriocheir sinensis; Yang et al., 2015; Liu, 2016). More research further revealed the harmful effects of this dinoflagellate on Ruditapes philippinarum, Brachionus plicatilis and Calanus sinicus (Yang et al., 2015; Liu, 2016). Microscopic examination of the blooming species indicated that it did not resemble any common dinoflagellates in this region, and the true species identity is yet to be determined. Here in this study, we reported the taxonomic research on this blooming species, which may assist future research on toxicity, physiology and assessment on ecological impact.
2 Materials and methods
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2.1 Sample collection
Vegetative cells were isolated from the aquaculture ponds in the coastal water of Panjin (40°51′N, 121°46′E) in 2008 and 2012. The original cultures were established by the Guanghe Crab Industry Limited Company (Panjin, China). The subset samples were then sent to the First Institute of Oceanography, State Oceanic Administration in Qingdao. They were maintained in f/2 medium with 0.50 mg/L GeO2 to inhibit diatom growing at 20 °C with an illumination of 100 μmol photons m–2 s–1 and 12L:12D photoperiod. The cells were transferred to the fresh medium every month to maintain the viability.
2.2 Light microscopy
Cell observation was carried out using a compound microscope (Olympus BX53, Japan) equipped with epifluorescence and differential interference contrast optics. In order to avoid the size reduction during cell fixation and dehydration process (Salas et al., 2014), we measured sizes of live cells. A drop of cultured cells were covered with a cover slip and observed at 1 000× magnification. After the cells slightly settled, images were captured and cell sizes were then measured using the CellSens Imaging software (Olympus, Japan). To examine the thecal plate, cells were fixed in 2% glutaraldehyde (final concentration), stained with Calcofluor White (Fritz and Triemer, 1985) and then checked under a violet excitation light. The shape and location of the chloroplasts were observed under a blue excitation light. The nucleus was stained with 4′-6-diamidino-2-phenylindole (DAPI, 0.1 μg/mL final concentration) and determined under a violet excitation light.
2.3 Electron microscopy
For SEM, cultured cells were either directly fixed by OsO4 or undergone a pretreatment process to clean the out layer debris before they were fixed and dehydrated. About 5 mL of cells at exponential growing phase were fixed with an equal volume of 4% OsO4 (w/v) for about 40 min. The fixed cells were then filtered onto a nylon membrane (5 μm pore-size) by gravity. Subsequently, the folded membrane with cells was washed by an acetone series (10%, 30%, 50%, 70%, 80%, 90%, 100% and repeat 100% for two times, critical point dried and coated with gold. For another batch of sample, a pretreatment process was performed to clean the out layer before they were fixed. Briefly, cells were collected by centrifugation at 5 000 r/min for 5 min, the supernatant was then removed and cell pellet was re-suspended in 60% ethanol at 4°C for 1 h to strip off the outer cell membrane. Subsequently, cells were pelleted again and re-suspended in 5 mL filtered seawater for 30 min at 4°C. These re-suspended cells were then fixed and dehydrated as described above. The gold-coated cells were viewed under a scanning electron microscope (Hitachi S-3400N, Japan). Some SEM micrographs were presented on a black background using Adobe Photoshop 6.0 (Adobe Systems, San Jose, California, USA).
For the transmission electron microscopy, fixed cells (2% OsO4) were dehydrated through an acetone series and then embedded in Spurr’s resin. Thin sections were cut with a diamond knife, stained with 2% uranyl acetate and observed using Hitachi 7 500.
2.4 PCR amplification and sequence analysis
Dinoflagellate cells were pelleted through centrifugation at 10 000 r/min for 5 min, cell pellet was then lysed and extracted using E.Z.N.A.TMHP plant DNA kits (OMEGA Bio-tek Inc., GA, USA) following the manufacturer’s protocol. PCR amplifications of ITS and LSU rRNA genes were performed as D’Onofrio et al. (1999) using the primer pairs: ITS1 5′-TCCGTAGGTGAACCTGCGG-3′, ITS4 5′-TCCTCCGCTTATTGATATGC-3′ (White et al., 1990) and D1R 5′-ACCCGCTGAATTTAAGCATA-3′, D2C 5′-CCTTGGTCCGTGTTTCAAGA-3′ (Edvardsen et al., 2003), respectively. Amplicons were ligated into pMD®18-T vectors (TaKaRa Co., Dalian, China) and then transformed into DH5∂ competent cells (TaKaRa) following the manufacture’s instructions. Selected clones were screened using M13 primers and those with right insertions were subsequently sequenced bi-directionally by the TsingKe Biological Technology (Qingdao, China).
The resulting sequences were cleaned for the ambiguous reads manually and then aligned with the Hetercapsa sequences retrieved from the public database (https://www.ncbi.nlm.nih.gov, up to date of April 7, 2017). Two sequences from Cochonina hallii (JQ972674 and AF033867) were also included in the analyses, given its unresolved taxonomic status relevant to Heterocapsa (Horiguchi, 1995). Sequences of Prorocentrum minimum (AF208244, AF260379) were treated as outgroups in ITS and LSU analyses, respectively. These sequences were aligned using MUSCLE algorithm (Edgar, 2004) implemented in MacVector 15.1.5 (Accelrys, CA, USA). Maximum-likelihood (ML), Neighbour-joining (NJ) and Maximum-parsimony (MP) analyses were performed in MEGA 6 (Tamura et al., 2013) with the best DNA substitution models (Kimura 2-parameter for ITS and Tamura-Nei for LSU) chosen based on Bayesian Information Criterion (BIC). Bootstrap analyses (1 000 replicates) were then conducted to evaluate the robustness of each clade. Gaps were deleted for the analyses.
3 Results
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3.1 Cell morphology
Cells are ellipsoidal, elongate with an average length of (14.4±1.6) μm (9.9–16.5 μm, n=102) and width of (10.2±1.4) μm (6.7–12.4 μm). The L:W ratios range from 1.2 to 1.8 with a mean of 1.4. The episome of cells ((8.4±2.1) μm in length, (10.2±1.4) μm in width) is significantly bigger than the hyposome ((4.1±1.1) μm, (8.2±1.2) μm, P<0.01, two-tailed t-test). Cingulum is wide and accounts for about 1/7 to 1/5 of the total cell length, evidently towards the antapex. The large nucleus is ellipsoid, mostly located in the middle of the cell (Figs 1a, b) and the numerous condensed chromosomes are visible when stained with DAPI (Fig. 1c). The spherical pyrenoids (1–3 per cell) are surrounded by starch sheaths, located above or beside the nucleus often in the episome or close to cingulum (Fig. 1c). A red accumulation body with variable shape and size is consistently observed in the hyposome (Figs 1a, b). The chloroplast is visible in the periphery of the cell (Fig. 1d) under the epifluorescence microscope. In the exponentially growing culture, cells are often divided by binary fissions (Figs 1e, f). Temporary cysts or an intermediate stage of planozygote and resting cyst are often observed at the bottom of the culture vessels and surrounded by heavy mucus. They are spherical to oval, significantly larger than the motile cells, and full of pale granules (Fig. 1g).
3.2 Thecal plate morphology
After being stained with Calcofluor White, the thecal plates of H. bohaiensis are visible (Fig. 1h) and consistent with the observations from SEM (Fig. 2). The plate pattern is schematized as in Fig. 3. Consistent with known Heterocapsa spp., the thecal plate configuration of H. bohaiensis is Po, cp, 5′, 3a, 7′′, 6C, 5s, 5′′′, 2′′′′. As shown in the diagram (Fig. 3), the epitheca consists 5 apical plates, 3 anterior intercalary plates and 7 precingular plates. The cingulum composes of 6 plates, and the hypotheca has 8 plates. The body scale (300–350 nm) is outlined by a triangle basal plate (Fig. 2e). A central upright and nine peripheral (three at each ridge) spines are visible on the basal plate. Three radiating spines (one at each side of the triangle) are probably connected with the central upright by the radiating bars. Interestingly, the morphology of apical pore complex (APC) varies when cells were treated with different methods. When cells are directly fixed with OsO4 without pretreatment, a spherical (0.8 μm in diameter) protruded cover plate (cp) is consistently observed above the pore plate (po, Fig. 2c), and there are visible radical rod-like decorations on cp. Whereas, when cells are pretreated using ethanol to strip off the outer layer, the cover plate (cp) turns to be flat, not protruded, and no decorations could be observed on the cp (Fig. 2d). The APC morphology of pretreated H. bohaiensis cells is similar to that of H. minima (Salas et al., 2014).
3.3 Ultrastructure
The ultrastructure of the cell shows a large dinokaryon, a presumably single chloroplast, a spherical pyrenoid, mitcondria, trichocysts and an accumulation body (Fig. 4a). The chloroplast with lamellae of three appressed thylakoids is located in periphery of the cell and connects with the pyrenoid. The pyrenoid is often surrounded by the starch sheath, and the thylakoids do not penetrate into the pyrenoid (Fig. 4b). A large accumulation body is quite visible in the hyposome.
3.4 Phylogeny of H. bohaiensis
The ITS sequences form about 15 distinct clades (Fig. 5) including the one representative of H. bohaiensis from this study. The eight monophyletic clades with high bootstrapping support values represent taxa of H. horiguchii, H. circularisquama, H. triquetra, H. illdefina, H. arctica, H. lanceolata, H. minima, H. pygmaea, respectively. Additional three single sequences (AB084098, KF240777 and AB445394) are genetically divergent with their sister taxa and formed three individual clades, standing for the species of H. ovata, H. rotundata and H. huensis, respectively. Interestingly, the sequences of FJ823556 and FJ823557, both designated as H. niei, fall into two distinct groups. The sequence FJ823556 (isolate UTEX 2722, Stern et al., 2012) is loosely clustered with AY499509 (Heterocapsa sp. GeoB 222, Gottschling et al., 2005) forming a sister clade to H. pseudotriquetra (AB084100), while the other (FJ823557, CS-36, Stern et al., 2012) is clustered with accessions FJ823557, JN020158 (H. niei identified in Iran; Attaran-Fariman and Javid, 2013) and JQ991005 (H. sp. CCMP424 collected from Australia) forming a strongly supported clade. Another well-supported clade (labeled as Heterocapsa sp. in Fig. 5) consists of four sequences (KX853194, KX853193, KX853191 and JQ9726681) with unknown identities, one (AB084093) designated as H. pygmaea (CCMP 1322), and one (JQ972674) as Cachonina hallii (CCMP 2770). The p-distances between H. bohaiensis and the closest lineages (H. pygmaea, H. sp., H. huensis and H. niei) are 0.059–0.087, which is comparable to the levels between H. pygmaea and H. huensis (0.099), H. rotundata and H. lanceolata (0.087), H. lanceolata and H. arctica (0.062), H. minima and H. arctica (0.095).
A total of 33 LSU sequences, including 28 retrieved from NCBI GenBank and 5 obtained in the study, form two reciprocally monophyletic clades (Fig. 6). Heterocapsa bohaiensis is located within the clade comprising H. pygmeae (EU165306), one undetermined strain (KT860562) isolated from the Mediterranean Sea, a few strains from the Gulf of Mexico, Florida of USA (FJ939577, EU165271 and EU165312), Iran (JN119844) and the Arabian Gulf of Turkey (KX853175, 77, 78, 87). The other clade with low bootstrapping support consists of the rest 18 sequences representing various strains from east coast of USA, the Baltic Sea, the Mediterranean Sea, and off the coasts of Australia, New Zealand, South Korea, etc.. Noticeably, both ITS and LSU sequences of Cachonina hallii (JQ972674, AF033867) are clustered within the Heterocapsa clades and show little sequence divergence (<0.1%) with the neighbor sequences. This indicates a problematic taxonomic status of Cachonina, which requires more research.
4 Discussion
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Taxonomy of Heterocapsa is difficult due to the small cell size and similarity of cellular morphology. The diagnostic characters included cell size and shape, shape and position of nucleus, number and position of pyrenoid, etc. (Iwataki et al., 2003; Iwataki, 2008). The newly found species described as Heterocapsa bohaiensis in this study is morphologically similar to H. pygmaea and H. huensis, both have ovoid cells (about 12–22 μm in length) and multiple pyrenoids. H. bohaiensis, however, posseses evidently larger epitheca compared to its hypotheca (Fig. 1), which is different from the almost equal size of epi-, hypotheca of H. pygmaea and H. huensis (Loeblich III et al., 1981; Iwataki et al., 2009). According to a review by Iwataki (2008), four species (H. arctica, H. lanceolata, H. minima and H. rotundata) reported to have a relatively larger epitheca, while they could be distinguished from H. bohaiensis by the cell size, shape and number of pyrenoids, as summarized in Table 1. Cells of H. arctica are quite elongated and were only observed in Arctic and the Baltic Sea where water temperatures are lower than 5°C (Horiguchi, 1997; Rintala et al., 2010). Heterocapsa lanceolata is featured by the unique lanceolate shapes (Horiguchi, 1997; Iwataki, 2008). Cells of H. minima and H. rotundata are relatively small (H. minima: 10–13 μm in length, mean=11.8 μm, Salas et al., 2014; H. rotundata: 9–12 μm, 10.5 μm, Hansen, 1995). All the four species have only one pyrenoid per cell (Iwataki, 2008; Rintala et al., 2010; Salas et al., 2014), which is distinct from H. bohaiensis (1–3 pyrenoids). The body scales have long been recognized present on the cell surface of Heterocapsa. Iwataki et al. (2004) compared and summarized the ultrastructure of body scales for 12 Heterocapsa species, and suggested its utility in the taxonomy of Heterocapsa spp. (Tamura et al., 2005; Iwataki et al., 2004). Although we failed to reveal the detailed structure through TEM, observation under SEM indicated that body scales of H. bohaiensis are outlined by a triangle basal plate (300–350 nm). The central upright and 9 peripheral spines are present on the basal plate (Fig. 2e). The body scale of H. bohaiensis is morphologically similar to that of H. huensis (Iwataki et al., 2009). The sequence phylogeny of both ITS and LSU further supports that H. bohaiensis is a novel species divergent from other described Heterocapsa spp.. Therefore, we classified it as a new species with the formal description as following:
Heterocapsa bohaiensis Xiao and Li sp. nov.
(Figs 1–4)
Cellula ellipsoideae, 9.9–16.5 μm longae, et 6.7–12.4 μm latae; ex epitheca grandi et hypotheca parva constans; tabulatio thecalis Po, cp, 5′, 3a, 7′′, 6c, 5s, 5′′′, 2′′′′; Nucleus elongatus, in parte hypoconi ad mediam cellulae situs.; pyrenoides singularis ad tres, sphaerica, cum incursionibus cytoplasmatis et amylo cingente; haec squama structura similaris Heterocapsa huensis Iwataki.
Description: Cells are ellipsoidal, 9.9–16.5 μm long, 6.7–12.4 μm wide; epitheca larger than hypotheca; thecal plate arrangement po, cp, 5′, 3a, 7′′, 6c, 5s, 5′′′, 2′′′′; Nucleus elongated and centrally located; pyrenoids one to three, spherical, surrounded by starch sheaths. The structure of body scale is similar to that of Heterocapsa huensis Iwataki.
Holotype: Fig. 1.
Type locality: Panjin (40°51′N, 121°46′E), Liaodong Bay, Bohai Sea, China.
Etymology: The species name refers to the Bohai Sea, the innermost gulf in northern China, where this species was firstly detected and isolated.
As described above, the morphology of APC varied when cells were treated using different methods. A protruded cover plate with radical rod-like decorations was consistently observed for the cells directly fixed with OsO4. Similar protruded cover plate was observed in H. pygmaea, which was also fixed directly (Roberts et al., 1987). In contrast, there was a flat cover plate with no visible ornaments (Fig. 3) when cells of H. bohaiensis were pretreated. This morphology is consistent with the observation of H. minima (Salas et al., 2014), which were also pretreated similarly with ethanol. Apparently, the pretreatment procedure not only tripped off the outer layer of the dinoflagellate cells, but also could affect the appearance of the cover plate. Apical structure has been recognized as an important feature for the taxonomy of dinoflagellate (Dodge and Hermes, 1981; Toriumi and Dodge, 1993; Hoppenrath, 2017). Dodge and Hermes (1981) compared the apical pore structures of marine dinoflagellates, suggesting ornamentation and shape of cover plate may vary among taxa in certain genera. Unfortunately, there was few research reported the APC structure of Heterocapsa, even less the shape and ornament of the cover plate, so that we could not compare the APCs among Heterocapsa spp.. Given the distinct morphology of APCs among dinoflagellates (Dodge and Hermes, 1981; Toriumi and Dodge, 1993; Hoppenrath, 2017), more research is needed on the APCs of Heterocapsa, which may provide valuable information for intra- and inter-generic taxonomy studies.
So far, H. bohaiensis was only confirmed occurring in Panjin, the Bohai Sea, while the distribution of this species has not been fully investigated. Both ITS and LSU sequences reveal a monophyletic clade of H. bohaiensis, which differs from other Heterocapsa species. Based on the ITS sequences, the genetic distances between H. bohaiensis and its closely-related taxa H. huensis, H. pygmaea and H. niei are comparable to those among the sister taxa. The sequence phylogenetic analyses support the findings from the morphological data. A recent survey detected multiple ribosomal sequences with high identity (>99%) with SSU sequences of H. bohaiensis in the coastal water of Qinghuangdao, the Bohai Sea (data not shown), suggesting this species may distribute more broadly. In addition, blooms of H. circularisquama and H. rotundata were recorded in the Changjiang Estuary and the coast of Qingdao in 2003, 2005 and 2008 (Wang et al., 2006), although no informative morphological and sequence data were provided. Heterocapsa-like sequences were also detected in the South China Sea as indicated by the deposited sequence accessions (KT389857, KT389866, KT389965, KT389969, KT389981 and KT389989). These sequences, however, were not included in the phylogenetic analyses of this study due to their un-alignability of multiple gaps and insertions in these sequences (data not shown). Nonetheless, all the information above implied probably a high species diversity of Heterocapsa spp. off the coasts of China, and further research is needed to elucidate the diversity and distribution of Heterocapsa spp. in China, including the newly found H. bohaiensis.
Although the bloom of H. bohaiensis in natural environment was not reported so far, significant mortalities of cultured Penaeus japonicus and Eriocheir sinensis lavae were observed in aquaculture ponds with H. bohaiensis cell abundance of 105 cells/mL (salinity 26). More laboratory testing suggested the harmful effects of H. bohaiensis on various marine organisms. Preliminary tests showed that high density (2×105 cells/mL) of H. bohaiensis could hamper the reproduction of rotifer Brachionus plicatilis (the major predator of cultured Eriocheir sinensis larvae) and metamorphosis of Eriocheir sinensis zoea (Yang et al., 2015). The mortality (cumulative mortality of 98.3%) of testing Manila clams (Ruditapes philippinarum) cultured with H. bohaiensis was significantly higher than that of the control groups (14.2% mortality, cultured with Chlorella vulgaris, Liu, 2016). In one toxicity screening, about 0.37 μg/g GTX4 (one component of PSP toxins) was detected from the Ruditapes philippinarum tissues fed with H. bohaiensis (Liu, 2016), while no toxins (e.g., PSP) were identified in the cultured H. bohaiensis cells after multiple trials (Yang et al., 2015; Gao Chunlei, personal communication). Furthermore, no tests have been conducted on the photosensitizing hemolytic toxin which has been recently isolated from H. circularisquama (Sato et al., 2002; Miyazaki et al., 2005), a red tide dinoflagellate harmful to bivalves in Japan (Horiguchi, 1995; Matsuyama et al., 1995, 1997, 2001). Apparently, more research is needed to clarify the harmful effect or toxicity of H. bohaiensis.
Acknowledgements
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The authors gratefully acknowledge Hu Zhangxi, Liu Wei from the Insititute of Oceanology, Chinese Academy of Science for their assistance on EM, and Miao Xiaoxiang and Ge Meiling for maintaining the culture.
Funding
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  • The NSFC-Shandong Joint Funded Project under contract No. U1406403; the National Natural Science Foundation of China under contract Nos 41506191 and 41306171.
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Appendix
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Year 2018 volume 37 Issue 10
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doi: 10.1007/s13131-018-1296-z
  • Receive Date：2017-08-01
  • Online Date：2026-04-14
  • Published：2018-10-25
Article Data
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History
  • Received：2017-08-01
  • Accepted：2018-02-13
Funding
The NSFC-Shandong Joint Funded Project under contract No. U1406403; the National Natural Science Foundation of China under contract Nos 41506191 and 41306171.
Affiliations
    1 Key Laboratory of Science and Engineering for Marine Ecology and Environment, the First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China
    2 Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China
    3 Guanghe Crab Industry Limited Company, Panjin 124200, China
    4 School of Food and Environment, Dalian University of Technology, Panjin 124221, China

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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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