The morphology of the two marine urostyloid ciliates,
Kahl (1932) established the genus
In this study, we described two marine ciliates new to Korea,
The specimens of
After collection and isolation, specimens were maintained in the laboratory, either as pure or raw cultures in Petri dishes and 50 mL tissue culture flasks (Greiner Bio-one, Fricken-hausen, Germany). Autoclaved seawater was supplied with putting rice grains as a substrate for bacterial growth (Jung et al., 2011). The living specimens were observed under a light microscope ( Leica DM2500; Leica Microsystems, Wetzlar, Germany) at 50-1,000 magnification. Protargol impregnation was applied according to Foissner (1991) to reveal the infraciliature.
Terminology and classification are mostly according to Berger (2006) and Lynn (2008).
A cell (single specimens of each species) was transferred to a 1.5 mL microtube with a minimum volume of water. Geno-mic DNAs were extracted using a RED-Extract-N-Amp Tis-sue PCR kit (Sigma, St. Louis, MO, USA), according to the manufacturer’s protocol. The nearly complete SSU rRNA genes were amplified by polymerase chain reaction (PCR) with the universal eukaryotic primers: New EukA (5？-CTG GTT GAT YCT GCC AGT-3？), modified from Medlin et al. (1988), and LSU rev3 (Sonnenberg et al., 2007) primers. The optimized conditions for this process were as follows: Denaturation at 94℃ for 3 min followed by 35 cycles of denatura-tion at 95℃ for 15 sec, annealing at 58℃ for 30 sec, extension at 72℃ for 4 min, and then a final extension step at 72℃ for 7 min. The PCR products were purified with the QIAquick® PCR Purification kit (Qiagen, Valencia, CA, USA). Three internal primers were used for sequencing: 18S+810 (5？-GCC GGA ATA CAT TAG CAT GG-3？) and 18S-300 (5？-CAT GGT AGT CCA ATA CAC TAC-3？) and 18S+1470(5？-TCT GTG ATG CCC TTA GAT GTC-3？). Sequencing in both directions was conducted by an ABI 3700 Sequencer(Applied Biosystems, Foster City, CA, USA).
The sequencing fragments of the SSU rRNA gene were combined via BioEdit (Hall, 1999) and were aligned using Clustal X 1.81 (Jeanmougin et al., 1998). Mega 4.0 (Tamura et al., 2007) was used to calculate genetic distance by applying the Kimura two-parameter distance method (Kimura, 1980).
Korean name: 1*뚱뚱이홍색위각모충 (신칭)
Phylum Ciliophora Doflein, 1901
Class Spirotrichea Butschli, 1889
Order Urostylida Jankowski, 1979
Family Pseudokeronopsidae Borror and Wicklow, 1983
Genus Pseudokeronopsis Borror and Wicklow, 1983
1*Pseudokeronopsis carnea (Cohn, 1866)
Wirnsberger et al., 1987 (Table 1, Figs.1 A-D, 2)
[Fig. 1.] Morphology and infraciliature of Pseudokeronopsis carnea and Uroleptopsis citrina from live specimens (A B E F) and afterprotargol impregnation (C D G H). A-D Pseudokeronopsis carnea: A Ventral view of live specimen arrowhead in (A) denotes CV;B Two types of granules; infraciliature of the ventral (C) and dorsal (D) sides. E-H Uroleptopsis citrina: E Ventral view of livespecimen; F Two types of granules; infraciliature of the ventral (G) and dorsal (H) sides. AZM adoral zone of membranelles; BCbuccal cirri; CV contractile vacuole; DK dorsal kineties; EM endoral membrane; FTC frontoterminal cirri; LMR left marginal row;Ma macronuclei; MVR midventral row; PM paroral membrane; RMR; right marginal row; TC transverse cirri. Scale bars=100 μm.
[Fig. 2.] Morphology and infraciliature of Pseudokeronopsis carnea from live specimens (A-C E-G) and after protargol impregnation(D H-L). A B Ventral views of live specimen; C Dorsal views of live specimen arrow marks a contractile vacuole; E Cortical granulesaround dorsal kineties; Arrows in (F G) indicate ring-shaped hollow structures arrowheads show cortical granules; D H-K Ventraland (L) dorsal views of protargol-impregnated specimen; D General ciliature of the specimen; H Frontal (bicorona) arrow indicatesthe buccal cirrus; I Two frontoterminal cirri arrows indicate the cirri near the distal end of adoral zone; J Two midventral rows; KDenotes transverse ventral cirri; Arrows in (L) show the dorsal kineties. Scale bars=100 μm.
1987, Song et al., 2006: 271-287, figs.1 A-G, 2, 3, 9C, Tables .1-3
The adoral zone of membranelles distinct, approximately 1/3 of the cell length, and composed of about 69 membran-elles(Fig.2 D, H). Bicorona of frontal cirri slightly enlarged,composed of about 8-12 cirral pairs, extending as a midven-tral complex consecutively. One buccal cirrus near the paro-ral membrane (Fig.2 H, arrow), whereas two frontoterminal cirri behind the distal end of the adoral zone (Fig.2 I, arrows); midventral complex distinctly separated rows (Fig.2 J), com-posed of 30-46 cirral pairs, terminating near transverse cirri; both posterior ends of marginal cirral rows not overlapped; 7-10 transverse cirri located between both posterior ends of the left and right marginal cirral rows (Fig.2 K). Almost no gap found between the midventral rows and the transverse cirri; from five to seven dorsal kineties (Figs.1 D; 2L, arrows).
Eight species among the genus
The Korean population,
[Fig. 3.] Morphology and infraciliature of Uroleptopsis citrina from live specimens (A B E F) and after protargol impregnation (C DG-I). A B Ventral views of live specimen; C Dorsal and (D G-I) ventral views of protargol-impregnated specimen; C Arrows markthe invariable three dorsal kineties; C D Arrowheads point to gap in adoral zone; D General ciliature of the specimen; E F Arrowand arrowheads indicate the two kinds of granules respectively; G Anterior pairs and single cirri (arrow mark) on the midventralcomplex; H Indicates macronucleus; I Frontal cirri (bicorona); arrows show two frontoterminal cirri. Scale bars=100 μm.
The adoral zone of membranelles distinct; about 1/3 of cell length, and composed of about 40 membranelles (Fig.3 D, I), left anterior corner a minute process causing a break (Fig.3 C, D, arrowhead). Bicorona of frontal cirri slightly enlarged, composed of about 6-9 cirral pairs, extending as a midventral complex consecutively (Fig.3 I). Midventral complex distin-ctly separated rows, composed of 26-35 cirri containing anterior, single cirri (Fig.3 G, arrow) in middle portion, posterior portion. Two or three frontoterminal cirri behind the distal end of the adoral zone (Fig.3 I, arrows); invariably three dorsal kineties (Figs.1 H; 3C, arrows); of particular interest, there is no buccal cirrus and transverse cirri.
lows: presence of a buccal cirrus and the pattern of the mid-ventral complex. Circumstantially,
The Korean population,