First Report of Heterocapsa minima
(Dinophyceae) from Aotearoa/
New Zealand
Small planktonic armoured dinoflagellates within the genus Heterocapsa are
currently represented by 20 species
with some having a world-wide distribution [1-2]. Blooms of some Heterocapsa species have been associated
with fish kills due to oxygen deficiency
or oversaturation. Toxicity by H. circularisquama has also been reported, with
blooms causing tremendous damage to
bivalve species [3-4]). Heterocapsa triquetra is one of the most common red
tide species around the world [1]. Recently, Tillmann et al. [2] proposed a
new species, H. steinii, for a taxon that
has been known as H. triquetra [5]. It is
therefore important to elucidate the diversity and distribution of Heterocapsa
species and to understand their potential adverse impacts in coastal areas.
In Aotearoa/New Zealand, a nearshore red-tide bloom of a Heterocapsa
species at Pukehina Beach, Bay of Plenty on 10 December 2003 was associated with swimmers complaining of skin
irritations [6] (Site 3 in Fig. 1). Previous
studies identified and reported four
Heterocapsa species from New Zealand
based on the results of morphological
characterisations of clonal strains (Sites
1, 2, and 3 in Fig. 1) [7]: H. cf. circularisquama, H. illdefina, and H. niei from
the subtropical zone [6,8] and H. steinii
(previously H. triquetra) from the temperate zone [7]. However, the diversity
of Heterocapsa species in the temperate
zone of New Zealand has not been well
elucidated.
In the present study, seawater samples were collected from two sampling
sites from the South Island and three
sampling sites from Stewart Island of
New Zealand which are within the temperate zone (Fig. 1): Site 4 (Wedge Point,
Marlborough; -41.2578, 174.0116) on
12 December 2018, Site 5 (Akaroa, Canterbury; -43.8048, 172.9653) on 18 October 2018, Sites 68 [Site 6 (Halfmoon
Bay; -46.8968, 168.1305), Site 7 (Golden Bay Wharf; -46.9041, 168.1216),
and Site 8 (Big Glory Bay; -46.9793,
168.1099), Stewart Island, Southland]
on 02 September 2018. The samples
were collected by hose sampling at
8
Fig. 1. Summary of the geographic distribution of Heterocapsa species in coastal areas of
New Zealand. Circles represent the sampling sites of clonal strains (white: previous
studies, grey: previous and present studies,
dark grey: the present study). 1[8], 2[6], 3[7],
4The present study. 1,2 Strains of H. niei and H.
illdefina were identified by electron microscope examination of thecal plate structure and
scale morphology. A strain of H. cf. circularis
quama was identified by electron microscope
examination of thecal plate structure alone.
3A strain of H. steinii was identified by light
microscope examination alone. 4Strains of H.
minima and H. steinii were identified by ribosomal RNA gene sequencing. Site 1: Kerikeri,
Northland. Site 2: Bream Bay, Northland.
Site 3: Pukehina Beach, Bay of Plenty. Site 4:
Wedge Point, Marlborough. Site 5: Akaroa,
Canterbury. Sites 68: Site 6, Halfmoon Bay;
Site 7, Golden Bay Wharf; Site 8, Big Glory
Bay; Stewart Island, Southland. The potential
boundary between the subtropical and temperate zones is reported by National Institute
of Water and Atmospheric Research [15].
015 m depths at Site 4 and from surface seawater at Sites 58. Heterocapsa
cells were observed under an inverted
light microscope (LM), isolated in f/2
medium to establish clonal strains, and
incubated at 18 C and 90 μmol photons m2s1 under a 12:12 L/D cycle.
A total of 16 clonal strains were established: one from Site 4, four from Site 5,
and three from Site 6 (Halfmoon Bay),
three from Site 7 (Golden Bay Wharf),
and five from Site 8 (Big Glory Bay) in
Stewart Island. The cells of the strain
from Site 4 were smaller than those
from Sites 58 (data not shown). The
cells of strains from Sites 58 showed
the same morphology under LM (data
not shown). Therefore, three representative strains were selected for molecular phylogenetic identifications: strains
CAWD302 (Site 4), CAk01H (Site 5), and
SSB03H (Site 8). Genomic DNA of the
three strains was extracted, and the D1/
D3 region of the large subunit ribosomal RNA gene (LSU rDNA) was amplified
and sequenced as previously described
[9]. Molecular phylogenetic analyses
were conducted using maximum likelihood (ML) and neighbour-joining (NJ)
methods.
The phylogenetic analyses revealed
that two strains, CAk01H and SSB03H,
belonged to the species H. steinii (Fig.
2). The sequences of these two strains
were identical to those of strain
UTKG7 (GenBank accession numbers:
MF423357 and MF423361) that corresponds to the type material of this species. Strain CAWD302 belonged to species H. minima (Fig. 2). The sequence
of CAWD302 was identical to those of
strains JK2 and HMMJ1604 (GenBank
accession numbers: KF031312 and
MK483261, respectively).
Previous studies in New Zealand
have reported three Heterocapsa species (H. cf. circularisquama, H. illdefina,
and H. niei) from the subtropical zone
[6,8]. The present study has revealed
presence of two additional species: H.
minima from Site 4 and H. steinii from
Sites 5 and 8 in the temperate zone. Using molecular phylogenetic methods,
the present study supports the previous finding of H. steinii (previously H.
triquetra), identified by morphology as
described above, from Site 5 [7] (Fig.
2). The present study is the first report
of H. minima from New Zealand. Furthermore, it indicates that Heterocapsa
species composition may be different in
the subtropical and temperate zones of
New Zealand (Fig. 2).
Regarding the distribution of H. minima, this species was first described by
morphological characterization without molecular phylogenetic data from
the Celtic Sea in the temperate North
Atlantic Ocean in 1989 [10]. In 2001,
abundance data of H. minima were reported in the fixed samples collected
from the Bay of Biscay, North Spain in
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