A scaleless scale worm: Molecular evidence for the phylogenetic placement of Pisione remota (Pisionidae, Annelida)

Figures & data

Table I List of taxa, 18S data, and COI data. Accession numbers of determined sequences in bold (Smithsonian Museum voucher number and locality given for taxa from which we collected data).

			Accession number
Taxon	Locality	Voucher No.	18S	COI
Aphroditidae
Aphrodite negligens	Griffin Bay, off San Juan Island, WA, USA; 48° 28.260′N 122° 59.241′W	USNM 1077202	AY894294	AY894309
Aphrodite sp.	Monterey Bay, Station 1 at 30 m, CA, USA; 36° 51.433′N 121° 51.438′W	USNM 1077203	AY894295	AY894310
Glyceridae
Glycera americana			U19519
Hesionidae
Kefersteinia cirrata	Banyuls-sur-Mer, France	USNM 10772228	AY527052
Pholoidae
Pholoe pallida	near Tautra at 200–220 m, Trondheim, Norway; 63° 36.91′N 10° 40.07′E	USNM 1077214	AY894302	AY894318
Phyllodocidae
Anaitides sp.	Point Sur, Station 12.1 at 63 m, CA, USA; 36° 55.182′N 121° 54.279′W	USNM 1077215	AY894293	AY894308
Eteone longa			AF448155
Eteone picta	Roscoff, France	USNM 1077229	AY525626
Eumida sp.	Point Sur, Station 14 at 92 m, CA, USA; 36° 49.847′N 122° 01.729′W	USNM 1077216	AY894296	AY894311
Eulalia viridis			AY525627
Phyllodoce sp.			AB106249
Sige sp.	Point Sur, Station 9.2 at 107 m, CA, USA; 36° 23.381′N 121° 57.974′W	USNM 1077217	AY894305	AY894320
Pilargidae
Ancistrosyllis sp.			AF474280
Pisionidae
Pisione remota	Cefalù, Sicilia, Italy	USNM 1077230	AY525628	AF221575
Polynoidae
Alentia gelatinosa	Concarneau, France	USNM 1077231	AY525630
Gattyana ciliata	San Juan Channel, WA, USA; 48° 34.231′N 123° 02.247′W	USNM 1077218	AY894297	AY894312
Halosydna brevisetosa	Munlo Cove, Griffin Bay, WA, USA; 48° 29.513′N 123° 01.099′W	USNM 1077219	AY894298	AY894313
Harmothoë impar			U50968
Harmothoë oculinarum	Rødberg at 200 m, Trondheim, Norway; 63° 28.36′N 10° 00.04′E	USNM 1077220	AY894299	AY894314
Lepidonotus squamatus	Munlo Cove, Griffin Bay, WA, USA; 48° 29.513′N 123° 01.099′W	USNM 1077221	AY894300	AY894316
Lepidonotus sublevis	Southern New England, Station 1 at 33 m, MA, USA; 41° 09.869′N 70° 25.0405′W	USNM 1077222	AY894301	AY894317
Paralepidonotus ampulliferus			AF519237	AY583698
Nephtyidae
Nephtys hombergii			U50970
Nephtys hombergii	Concarneau, France	Completely used	AY527054
Nereididae
Ceratonereis longiceratophora			AB106251
Nereis limbata			U36270
Nereis pelagica			AF474279
Platynereis dumerilii	Seahorse Key dredge, FL, USA;29° 05.415′ N 083° 04.268′W	USNM 1077223	AY894303	NC_000931: 808756
Sigalionidae
Leanira sp.	Southern New England, Station 4 at 96 m, MA, USA; 40° 20.410′N 70° 46.765′W	USNM 1077224		AY894315
Sigalion bandaensis			AB106254
Sigalion spinosa	Point Sur, Station 9.3 at 107 m, CA, USA; 36° 22.550′N 121° 59.010′W	USNM 1077225	AY894304	AY894319
Sthenalanella uniformis	Monterey Bay, Station 1 at 30 m, CA, USA; 36° 51.433′N 121° 51.438′W	USNM 1077226	AY894306	AY894322
Sthenelais boa	Blacks Beach, MA, USA; 41° 35.191′N 70° 38.665′W	USNM 1077227	AY894307	AY894321
Syllidae
Autolytus prolifer			AF474295
Brania sp.	Roscoff, France	USNM 1077232	AY525633
Exogone naidina			AF474290
Typosyllis armillaris			AF474292
Amphinomidae
Paramphinome jeffreysi			AY838856	AY838875

Figure 1.  Relative position of 18S rDNA primers corresponding to residues of Homo sapiens.

Table II Amplification and sequencing primers. Positions correspond to residues of Homo sapiens (18S) and Platynereis dumerilii (COI).

Name	Sequence (5′→3′)	Position	Direction	Reference
18S
18e	CTG GTT GAT CCT GCC AGT	3–21	forward	Hillis & Dixon 1991
18F509	CCC CGT AAT TGG AAT GAG TAC A	548–569	forward	Struck et al 2002
18L	GAA TTA CCG CGG CTG CTG GCA CC	609–632	reverse	Halanych et al. 1995
18R925D	GAT CYA AGA ATT TCA CCT CT	955–974	reverse	Burnette et al. 2005
18F997	TTC GAA GAC GAT CAG ATA CCG	1044–1065	forward	Struck et al. 2002
18r	GTC CCC TTC CGT CAA TTY CTT TAA G	1191–1215	reverse	Passamaneck et al. 2004
18F1435	AGG TCT GTG ATG CCC TTA GAT	1489–1509	forward	Burnette et al. 2005
18R1779	TGT TAC GAC TTT TAC TTC CTC TA	1811–1834	reverse	Struck et al. 2002
18R1843	GGA TCC AAG CTT GAT CCT TCT GCA GGT TCA CCT AC	1843–1877	reverse	Modified from Cohen et al. 1998
COI
LCO1490	GGT CAA CAA ATC ATA AAG ATA TTG G	14–38	forward	Folmer et al. 1994
HCO2198	TAA ACT TCA GGG TGA CCA AAA AAT CA	697–722	reverse	Folmer et al. 1994
CO1r	CCD CTT AGW CCT ARR AAR TGT TGN GG	1270–1295	reverse	Modified from Nelson & Fisher 2000

Figure 2.  Plot T_i/T_v ratio against uncorrected distance p of both 1st and 2nd positions and 3rd positions of the COI data set.

Figure 3.  18S data set. ML phylogram based on TRNef + I+Γ (-ln L = 8,061.57). Only BS and PP above 50 or 0.50 shown, respectively. Settings-Base frequencies: A, C, G, T = 0.2500; Substitution rates: AG = 2.7990, CT = 4.1055, AC, AT, CG, TG = 1.0000; α = 0.678; pinvar = 0.4504.

Figure 4.  COI data set. A) ML phylogram based on TRN + Γ (-ln L = 1,715.69). Only BS above 50 shown. Settings-Base frequencies: A = 0.1997, C = 0.2650, G = 0.2350, T = 0.3003; Substitution rates: AG = 2.1121, CT = 13.5600, AC, AT, CG, TG = 1.0000; α = 0.0948. B) 50% majority rule consensus tree of BI analyis using GTR + Γ. Only PP above 0.50 shown.

Figure 5.  Combined data set. ML phylogram based on GTR + I+Γ (-ln L = 9,665.87). Only BS and PP above 50 or 0.50 shown, respectively. Settings-Base frequencies: A = 0.2685, C = 0.2066, G = 0.2297, T = 0.2952; Substitution rates: AC = 2.1036, AG = 5.2549, AT = 4.2558, CG = 1.7810, CT = 10.4237, TG = 1.0000; α = 0.3692; pinvar = 0.5587.

Hillis , DM and Dixon , MT . 1991 . Ribosomal DNA: Molecular evolution and phylogenetic inference . Quarterly Review of Biology , 66 : 411 – 53 .

 PubMed Web of Science ®Google Scholar

Halanych , KM , Bacheller , JD , Aguinaldo , AMA , Liva , SM , Hillis , DM and Lake , JA . 1995 . Evidence from 18S ribosomal DNA that the lophophorates are protostome animals . Science , 267 : 1641 – 3 .

 PubMed Web of Science ®Google Scholar

Burnette , A , Struck TH , Halanych KM . 2005 . Holopelagic Poeobius meseres (Poeobiidae, Annelida) is derived from within benthic Flabelligerid worms . Biological Bulletin. 208: in press .

 Google Scholar

Passamaneck , YJ , Schander , C and Halanych , KM . 2004 . Investigation of molluscan phylogeny using large-subunit and small-subunit nuclear rRNA sequences . Molecular Phylogenetics and Evolution , 32 : 25 – 38 .

 PubMed Web of Science ®Google Scholar

Cohen , BL , Gawthrop , A and Cavaliersmith , T . 1998 . Molecular phylogeny of brachiopods and phoronids based on nuclear-encoded small subunit ribosomal RNA gene sequences . Philosophical Transactions of the Royal Society of London. Series B: Biological Sciences , 353 : 2039 – 61 .

 Web of Science ®Google Scholar

Folmer , O , Black , M , Hoeh , W , Lutz , R and Vrijenhoek , R . 1994 . DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates . Molecular Marine Biology and Biotechnology , 3 : 294 – 9 .

 PubMedGoogle Scholar

Nelson , K and Fisher , CR . 2000 . Speciation of the bacterial symbionts of deep-sea vestimentiferan tube worms . Symbiosis , 28 : 1 – 15 .

 Web of Science ®Google Scholar