http://informahealthcare.com/mdn ISSN: 1940-1736 (print), 1940-1744 (electronic) Mitochondrial DNA, Early Online: 1–2 ! 2014 Informa UK Ltd. DOI: 10.3109/19401736.2013.855739
MITOGENOME ANNOUNCEMENT
Complete mitochondrial genome of the acorn barnacle Striatobalanus amaryllis (Crustacea: Maxillopoda): the first representative from Archaeobalanidae Ling Ming Tsang1*, Xin Shen2,3*, Ka Hou Chu2 and Benny Kwok Kan Chan4
Mitochondrial DNA Downloaded from informahealthcare.com by Universitat de Girona on 03/27/15 For personal use only.
1
Institute of Marine Biology, National Taiwan Ocean University, Keelung, Taiwan, 2Simon F. S. Li Marine Science Laboratory, School of Life Sciences, Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China, 3Jiangsu Key Laboratory of Marine Biotechnology, College of Marine Science, Huaihai Institute of Technology, Lianyungang, China and 4Biodiversity Research Center, Academia Sinica, Taipei, Taiwan
Abstract
Keywords
The mitochondrial genome of the barnacle Striatobalanus amaryllis (Sessilia: family Archaeobalanidae) is 15,063 bp in length. All the 13 protein-coding genes (PCGs) initiate with ATD codon (ATG, ATA or ATT). Four PCGs (COX3, ND3, ND4 and ND4L) end with incomplete stop codon (T- -). Four PCGs (ND1, ND4, ND4L and ND5) are encoded on the light strand (underlined below). Refer to the pancrustacean ground pattern, there are not less than seven tRNAs rearranged in the S. amaryllis mitochondrial genome, including tRNAAla, tRNAGlu/ tRNASer(AGY), tRNAPro/tRNAThr, tRNAPro/tRNAThr, tRNATyr, tRNALys, tRNAGln and tRNACys. Three tRNAs (tRNALys, tRNAGln and tRNACys) are rearranged between S. amaryllis and Tetraclita japonica (Sessilia: Tetraclitidae), meanwhile one tRNA (tRNACys) inverted from one strand to another. Compared with Megabalanus volcano (Sessilia: Balanidae), an inversion of one large gene block is identified (including three PCGs and three tRNAs) in S. amaryllis mitochondrial genome: tRNAPhe-ND5-tRNAHis-ND4-ND4L-tRNAPro.
Archeobalanidae, Balanoidea, Balanomorpha, Cirripedia, complete mitochondrial genome, Crustacea, gene order, Maxillopoda
The sessilian barnacle family Archeobalanidae (Crustacea: Cirripedia: Balanomorpha) is a diverse group including both free-living species and obligate symbionts of various marine taxa (Chan et al., 2009; Van Syoc & Newman, 2010). This study presents the complete mitochondrial genome of Striatobalanus amaryllis (collected from low intertidal rocks at Penang, Malaysia), which is the first report from the family Archeobalanidae. The mitochondrial genome is 15,063 bp in length and encodes 37 genes, including 13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNAs) genes and 22 transfer RNA (tRNAs) genes typical to metazoan mitogenome (Accession number: KF493890). All of the 13 PCGs in S. amaryllis mitochondrial genome initiate with ATG, ATA or ATT. This corroborates with the observation that metazoan mitochondrial protein-coding genes (PCGs) commonly use ATN as start codons (Shen et al., 2009a, 2012b). Only four PCGs (COX3, ND3, ND4 and ND4L) end with incomplete stop codon (T- -), with the remaining ones having complete stop codons (TAA or TAG; Table 1). Four PCGs (ND1, ND4, ND4L and ND5) are encoded on the light strand while the other 9 PCGs are located on the heavy strand. Both lrRNA and srRNA are encoded on the light strand, which is similar to other studied crustacean mitochondrial genomes (Shen et al., 2010, 2011, 2012a). Detailed genes contents of S. amaryllis mitochondrial genome are listed in Table 1.
*These authors contributed equally to this work. Correspondence: Benny Kwok Kan Chan, Biodiversity Research Center, Academia Sinica, Nankang, Taipei 115, Taiwan. Tel: +886 2 2789 9621. Email: [email protected]
History Received 12 September 2013 Revised 18 September 2013 Accepted 21 September 2013 Published online 7 January 2014
Compared with the pancrustacean ground pattern (Shen et al., 2007, 2009b), seven conserved gene blocks are found, including COX1-tRNALeu-UUR-COX2, tRNAAsp-ATP8-ATP6-COX3-tRNAGlyND3, tRNAArg-tRNAAsn, tRNAPhe-ND5-tRNAHis-ND4-ND4L, ND1-tRNALeu-CUN-lrRNA-tRNAValND6-CYTB-tRNASer-UCN, Ile Met srRNA-tRNA and tRNA -ND2-tRNATrp (underlined genes are encoded on the light strand, same below). There are not less than seven tRNAs rearranged in the barnacle S. amaryllis mitochondrial genome, including tRNAAla, tRNAGlu/tRNASer(AGY), tRNAPro/ tRNAThr, tRNATyr, tRNALys, tRNAGln and tRNACys (Table 1). The balanomorph acorn barnacles are a diverse group of crustacean (Martin & Davis, 2001; Newman & Ross, 1976), but only two mitochondrial genomes from the suborder Balanomorpha are available so far: Megabalanus volcano (Balanidae) (NC_006293) and Tetraclita japonica (Tetraclitidae) (NC_008974) (Begum et al., unpublished). Comparison of these two mitogenomes with this article reveals that mitochondrial gene arrangements are not conserved within the suborder. Three tRNAs (tRNALys, tRNAGln and tRNACys) are rearranged between S. amaryllis and T. japonica mitochondrial genomes. Meanwhile, one tRNA (tRNACys) inverted from one strand to another. Compared to M. volcano mitochondrial genome, gene rearrangements are not only restricted to tRNA genes, but include PCGs as well, also an inversion of one large genes block is found (including three PCGs and three tRNAs). In S. amaryllis mitochondrial genome, the order of six genes is: tRNAPhe-ND5-tRNAHis-ND4ND4L-tRNAPro, however, a new gene arrangement (tRNAProND4L-ND4-tRNAHis-ND5-tRNAPhe) is observed in M. volcano mitochondrial genome. Mitochondrial gene order is not conserved within the order Sessilia and represents a potential informative marker for inferring phylogenetic relationships within the order.
2
L. M. Tsang et al.
Mitochondrial DNA, Early Online: 1–2
Table 1. Gene profile of S. amaryllis (Crustacea: Maxillopoda) mitochondrial genome. Position
Mitochondrial DNA Downloaded from informahealthcare.com by Universitat de Girona on 03/27/15 For personal use only.
Gene COX1 tRNALeu-UUR COX2 tRNAAsp ATP8 ATP6 COX3 tRNAGly ND3 tRNAArg tRNAAsn tRNAAla tRNAGlu tRNASer-AGY tRNAPhe ND5 tRNAHis ND4 ND4L tRNAPro tRNAThr ND6 CYTB tRNASer-UCN tRNATyr tRNALys tRNAGln tRNACys ND1 tRNALeu-CUN lrRNA tRNAVal srRNA Control region tRNAIle tRNAMet ND2 tRNATrp
Codon
Strand
Start
Stop
Nucleotides (bp)
A þ T content (%)
Start
Stop
þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ
1 1547 1617 2377 2442 2594 3259 4046 4112 4464 4528 4593 4660 4726 4800 4863 6567 6639 7955 8244 8311 8374 8862 10,000 10,070 10,153 10,223 10,291 10,354 11,278 11,346 12,645 12,711 13,499 13,869 13,936 14,002 14,999
1545 1614 2300 2441 2600 3259 4045 4111 4463 4527 4592 4658 4725 4783 4863 6566 6630 7956 8234 8307 8373 8862 10,001 10,069 10,134 10,220 10,290 10,355 11,280 11,345 12,644 12,710 13,498 13,868 13,935 14,001 15,000 15,063
1545 68 684 65 159 666 787 66 352 64 65 66 66 58 64 1704 64 1318 280 64 63 489 1140 70 65 68 68 65 927 68 1299 66 788 370 67 66 999 65
65.8 69.1 70.2 73.8 77.4 75.1 67.5 72.7 72.2 50.0 81.5 71.2 69.7 63.8 70.3 74.3 71.9 72.9 73.6 67.2 65.1 75.7 67.7 61.4 63.1 75.0 72.1 67.7 70.1 76.5 74.1 68.2 69.5 81.1 71.6 68.2 75.3 66.2
ATT
TAA
þ þ þ þ
Anti-codon TAA
ATG
TAA
ATT ATG ATG
TAA TAA T- -
ATT
T- -
GTC
TCC CGC GTT TGC TTC GCT GAA ATG
TAA
ATG ATA
T- T- -
GTG TGG TGT ATG ATG
TAA TAG TGA GTA TTT TTG GCA
ATA
TAA TAG TAC GAT CAT
ATG
TAA TCA
Intergenic sequence* 1 2 76 0 7 1 0 0 0 0 0 1 0 16 1 0 8 2 9 3 0 1 2 0 18 2 0 2 3 0 0 0 0 0 0 0 2 0
*Negative values mean overlapping nucleotides.
Acknowledgements The authors thank Cheung Kwok Chu for his crucial comments, and the Genomics BioSci and Tech, Taiwan for supporting the sequencing of the mitochondrial DNA of the present study.
Declaration of interest The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the article. This study was supported by the Hong Kong Scholars Program (XJ2012056) and the Career Development Award in Academia Sinica, Taiwan (AS-98-CDAL15). Part of the study is supported by a grant in the National Science Council, Taiwan (NSC-99-2621-B-001-007-MY3).
References Chan BKK, Prabowo RE, Lee KS. (2009). Crustacean Fauna of Taiwan: Barnacles, Volume I – Cirripedia: Thoracica (Excluding the Pyrgomatidae and Acastinae). Keelung, Taiwan: National Taiwan Ocean University. Martin JW, Davis GE. (2001). An updated classification of the recent Crustacea. Los Angeles, CA: Natural History Museum of Los Angeles County. Newman WA, Ross A. (1976). Revision of the balanomorph barnacles, including a catalog of the species. San Diego: San Diego Society of Natural History. Shen X, Li X, Sha Z, Yan B, Xu Q. (2012a). Complete mitochondrial genome of the Japanese snapping shrimp Alpheus japonicus (Crustacea: Decapoda: Caridea): Gene rearrangement and phylogeny within Caridea. Sci China Life Sci 55:591–8.
Shen X, Ma X, Ren J, Zhao F. (2009a). A close phylogenetic relationship between Sipuncula and Annelida evidenced from the complete mitochondrial genome sequence of Phascolosoma esculenta. BMC Genom 10:136–46. Shen X, Ren J, Cui Z, Sha Z, Wang B, Xiang J, Liu B. (2007). The complete mitochondrial genomes of two common shrimps (Litopenaeus vannamei and Fenneropenaeus chinensis) and their phylogenomic considerations. Gene 403:98–109. Shen X, Sun M, Wu Z, Tian M, Cheng H, Zhao F, Meng X. (2009b). The complete mitochondrial genome of the ridgetail white prawn Exopalaemon carinicauda Holthuis, 1950 (Crustacean: Decapoda: Palaemonidae) revealed a novel rearrangement of tRNA genes. Gene 437:1–8. Shen X, Tian M, Meng X, Liu H, Cheng H, Zhu C, Zhao F. (2012b). Complete mitochondrial genome of Membranipora grandicella (Bryozoa: Cheilostomatida) determined with next-generation sequencing: The first representative of the suborder Malacostegina. Comp Biochem Physiol Part D Genom Proteom 7:248–53. Shen X, Wang H, Ren J, Tian M, Wang M. (2010). The mitochondrial genome of Euphausia superba (Prydz Bay) (Crustacea: Malacostraca: Euphausiacea) reveals a novel gene arrangement and potential molecular markers. Mol Biol Rep 37:771–84. Shen X, Wang H, Wang M, Liu B. (2011). The complete mitochondrial genome sequence of Euphausia pacifica (Malacostraca: Euphausiacea) reveals a novel gene order and unusual tandem repeats. Genome 54: 911–22. Van Syoc RJ, Newman WA. (2010). Morphology and evolutionary ecology of a sponge-barnacle symbiosis: Four new genera of barnacles (Archaeobalanidae, Bryozobiinae). J Exp Mar Bio Ecol 392:65–88.
MITOGENOME ANNOUNCEMENT
Complete mitochondrial genome of the acorn barnacle Striatobalanus amaryllis (Crustacea: Maxillopoda): the first representative from Archaeobalanidae Ling Ming Tsang1*, Xin Shen2,3*, Ka Hou Chu2 and Benny Kwok Kan Chan4
Mitochondrial DNA Downloaded from informahealthcare.com by Universitat de Girona on 03/27/15 For personal use only.
1
Institute of Marine Biology, National Taiwan Ocean University, Keelung, Taiwan, 2Simon F. S. Li Marine Science Laboratory, School of Life Sciences, Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China, 3Jiangsu Key Laboratory of Marine Biotechnology, College of Marine Science, Huaihai Institute of Technology, Lianyungang, China and 4Biodiversity Research Center, Academia Sinica, Taipei, Taiwan
Abstract
Keywords
The mitochondrial genome of the barnacle Striatobalanus amaryllis (Sessilia: family Archaeobalanidae) is 15,063 bp in length. All the 13 protein-coding genes (PCGs) initiate with ATD codon (ATG, ATA or ATT). Four PCGs (COX3, ND3, ND4 and ND4L) end with incomplete stop codon (T- -). Four PCGs (ND1, ND4, ND4L and ND5) are encoded on the light strand (underlined below). Refer to the pancrustacean ground pattern, there are not less than seven tRNAs rearranged in the S. amaryllis mitochondrial genome, including tRNAAla, tRNAGlu/ tRNASer(AGY), tRNAPro/tRNAThr, tRNAPro/tRNAThr, tRNATyr, tRNALys, tRNAGln and tRNACys. Three tRNAs (tRNALys, tRNAGln and tRNACys) are rearranged between S. amaryllis and Tetraclita japonica (Sessilia: Tetraclitidae), meanwhile one tRNA (tRNACys) inverted from one strand to another. Compared with Megabalanus volcano (Sessilia: Balanidae), an inversion of one large gene block is identified (including three PCGs and three tRNAs) in S. amaryllis mitochondrial genome: tRNAPhe-ND5-tRNAHis-ND4-ND4L-tRNAPro.
Archeobalanidae, Balanoidea, Balanomorpha, Cirripedia, complete mitochondrial genome, Crustacea, gene order, Maxillopoda
The sessilian barnacle family Archeobalanidae (Crustacea: Cirripedia: Balanomorpha) is a diverse group including both free-living species and obligate symbionts of various marine taxa (Chan et al., 2009; Van Syoc & Newman, 2010). This study presents the complete mitochondrial genome of Striatobalanus amaryllis (collected from low intertidal rocks at Penang, Malaysia), which is the first report from the family Archeobalanidae. The mitochondrial genome is 15,063 bp in length and encodes 37 genes, including 13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNAs) genes and 22 transfer RNA (tRNAs) genes typical to metazoan mitogenome (Accession number: KF493890). All of the 13 PCGs in S. amaryllis mitochondrial genome initiate with ATG, ATA or ATT. This corroborates with the observation that metazoan mitochondrial protein-coding genes (PCGs) commonly use ATN as start codons (Shen et al., 2009a, 2012b). Only four PCGs (COX3, ND3, ND4 and ND4L) end with incomplete stop codon (T- -), with the remaining ones having complete stop codons (TAA or TAG; Table 1). Four PCGs (ND1, ND4, ND4L and ND5) are encoded on the light strand while the other 9 PCGs are located on the heavy strand. Both lrRNA and srRNA are encoded on the light strand, which is similar to other studied crustacean mitochondrial genomes (Shen et al., 2010, 2011, 2012a). Detailed genes contents of S. amaryllis mitochondrial genome are listed in Table 1.
*These authors contributed equally to this work. Correspondence: Benny Kwok Kan Chan, Biodiversity Research Center, Academia Sinica, Nankang, Taipei 115, Taiwan. Tel: +886 2 2789 9621. Email: [email protected]
History Received 12 September 2013 Revised 18 September 2013 Accepted 21 September 2013 Published online 7 January 2014
Compared with the pancrustacean ground pattern (Shen et al., 2007, 2009b), seven conserved gene blocks are found, including COX1-tRNALeu-UUR-COX2, tRNAAsp-ATP8-ATP6-COX3-tRNAGlyND3, tRNAArg-tRNAAsn, tRNAPhe-ND5-tRNAHis-ND4-ND4L, ND1-tRNALeu-CUN-lrRNA-tRNAValND6-CYTB-tRNASer-UCN, Ile Met srRNA-tRNA and tRNA -ND2-tRNATrp (underlined genes are encoded on the light strand, same below). There are not less than seven tRNAs rearranged in the barnacle S. amaryllis mitochondrial genome, including tRNAAla, tRNAGlu/tRNASer(AGY), tRNAPro/ tRNAThr, tRNATyr, tRNALys, tRNAGln and tRNACys (Table 1). The balanomorph acorn barnacles are a diverse group of crustacean (Martin & Davis, 2001; Newman & Ross, 1976), but only two mitochondrial genomes from the suborder Balanomorpha are available so far: Megabalanus volcano (Balanidae) (NC_006293) and Tetraclita japonica (Tetraclitidae) (NC_008974) (Begum et al., unpublished). Comparison of these two mitogenomes with this article reveals that mitochondrial gene arrangements are not conserved within the suborder. Three tRNAs (tRNALys, tRNAGln and tRNACys) are rearranged between S. amaryllis and T. japonica mitochondrial genomes. Meanwhile, one tRNA (tRNACys) inverted from one strand to another. Compared to M. volcano mitochondrial genome, gene rearrangements are not only restricted to tRNA genes, but include PCGs as well, also an inversion of one large genes block is found (including three PCGs and three tRNAs). In S. amaryllis mitochondrial genome, the order of six genes is: tRNAPhe-ND5-tRNAHis-ND4ND4L-tRNAPro, however, a new gene arrangement (tRNAProND4L-ND4-tRNAHis-ND5-tRNAPhe) is observed in M. volcano mitochondrial genome. Mitochondrial gene order is not conserved within the order Sessilia and represents a potential informative marker for inferring phylogenetic relationships within the order.
2
L. M. Tsang et al.
Mitochondrial DNA, Early Online: 1–2
Table 1. Gene profile of S. amaryllis (Crustacea: Maxillopoda) mitochondrial genome. Position
Mitochondrial DNA Downloaded from informahealthcare.com by Universitat de Girona on 03/27/15 For personal use only.
Gene COX1 tRNALeu-UUR COX2 tRNAAsp ATP8 ATP6 COX3 tRNAGly ND3 tRNAArg tRNAAsn tRNAAla tRNAGlu tRNASer-AGY tRNAPhe ND5 tRNAHis ND4 ND4L tRNAPro tRNAThr ND6 CYTB tRNASer-UCN tRNATyr tRNALys tRNAGln tRNACys ND1 tRNALeu-CUN lrRNA tRNAVal srRNA Control region tRNAIle tRNAMet ND2 tRNATrp
Codon
Strand
Start
Stop
Nucleotides (bp)
A þ T content (%)
Start
Stop
þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ þ
1 1547 1617 2377 2442 2594 3259 4046 4112 4464 4528 4593 4660 4726 4800 4863 6567 6639 7955 8244 8311 8374 8862 10,000 10,070 10,153 10,223 10,291 10,354 11,278 11,346 12,645 12,711 13,499 13,869 13,936 14,002 14,999
1545 1614 2300 2441 2600 3259 4045 4111 4463 4527 4592 4658 4725 4783 4863 6566 6630 7956 8234 8307 8373 8862 10,001 10,069 10,134 10,220 10,290 10,355 11,280 11,345 12,644 12,710 13,498 13,868 13,935 14,001 15,000 15,063
1545 68 684 65 159 666 787 66 352 64 65 66 66 58 64 1704 64 1318 280 64 63 489 1140 70 65 68 68 65 927 68 1299 66 788 370 67 66 999 65
65.8 69.1 70.2 73.8 77.4 75.1 67.5 72.7 72.2 50.0 81.5 71.2 69.7 63.8 70.3 74.3 71.9 72.9 73.6 67.2 65.1 75.7 67.7 61.4 63.1 75.0 72.1 67.7 70.1 76.5 74.1 68.2 69.5 81.1 71.6 68.2 75.3 66.2
ATT
TAA
þ þ þ þ
Anti-codon TAA
ATG
TAA
ATT ATG ATG
TAA TAA T- -
ATT
T- -
GTC
TCC CGC GTT TGC TTC GCT GAA ATG
TAA
ATG ATA
T- T- -
GTG TGG TGT ATG ATG
TAA TAG TGA GTA TTT TTG GCA
ATA
TAA TAG TAC GAT CAT
ATG
TAA TCA
Intergenic sequence* 1 2 76 0 7 1 0 0 0 0 0 1 0 16 1 0 8 2 9 3 0 1 2 0 18 2 0 2 3 0 0 0 0 0 0 0 2 0
*Negative values mean overlapping nucleotides.
Acknowledgements The authors thank Cheung Kwok Chu for his crucial comments, and the Genomics BioSci and Tech, Taiwan for supporting the sequencing of the mitochondrial DNA of the present study.
Declaration of interest The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the article. This study was supported by the Hong Kong Scholars Program (XJ2012056) and the Career Development Award in Academia Sinica, Taiwan (AS-98-CDAL15). Part of the study is supported by a grant in the National Science Council, Taiwan (NSC-99-2621-B-001-007-MY3).
References Chan BKK, Prabowo RE, Lee KS. (2009). Crustacean Fauna of Taiwan: Barnacles, Volume I – Cirripedia: Thoracica (Excluding the Pyrgomatidae and Acastinae). Keelung, Taiwan: National Taiwan Ocean University. Martin JW, Davis GE. (2001). An updated classification of the recent Crustacea. Los Angeles, CA: Natural History Museum of Los Angeles County. Newman WA, Ross A. (1976). Revision of the balanomorph barnacles, including a catalog of the species. San Diego: San Diego Society of Natural History. Shen X, Li X, Sha Z, Yan B, Xu Q. (2012a). Complete mitochondrial genome of the Japanese snapping shrimp Alpheus japonicus (Crustacea: Decapoda: Caridea): Gene rearrangement and phylogeny within Caridea. Sci China Life Sci 55:591–8.
Shen X, Ma X, Ren J, Zhao F. (2009a). A close phylogenetic relationship between Sipuncula and Annelida evidenced from the complete mitochondrial genome sequence of Phascolosoma esculenta. BMC Genom 10:136–46. Shen X, Ren J, Cui Z, Sha Z, Wang B, Xiang J, Liu B. (2007). The complete mitochondrial genomes of two common shrimps (Litopenaeus vannamei and Fenneropenaeus chinensis) and their phylogenomic considerations. Gene 403:98–109. Shen X, Sun M, Wu Z, Tian M, Cheng H, Zhao F, Meng X. (2009b). The complete mitochondrial genome of the ridgetail white prawn Exopalaemon carinicauda Holthuis, 1950 (Crustacean: Decapoda: Palaemonidae) revealed a novel rearrangement of tRNA genes. Gene 437:1–8. Shen X, Tian M, Meng X, Liu H, Cheng H, Zhu C, Zhao F. (2012b). Complete mitochondrial genome of Membranipora grandicella (Bryozoa: Cheilostomatida) determined with next-generation sequencing: The first representative of the suborder Malacostegina. Comp Biochem Physiol Part D Genom Proteom 7:248–53. Shen X, Wang H, Ren J, Tian M, Wang M. (2010). The mitochondrial genome of Euphausia superba (Prydz Bay) (Crustacea: Malacostraca: Euphausiacea) reveals a novel gene arrangement and potential molecular markers. Mol Biol Rep 37:771–84. Shen X, Wang H, Wang M, Liu B. (2011). The complete mitochondrial genome sequence of Euphausia pacifica (Malacostraca: Euphausiacea) reveals a novel gene order and unusual tandem repeats. Genome 54: 911–22. Van Syoc RJ, Newman WA. (2010). Morphology and evolutionary ecology of a sponge-barnacle symbiosis: Four new genera of barnacles (Archaeobalanidae, Bryozobiinae). J Exp Mar Bio Ecol 392:65–88.
