Abstract
In order to improve the efficiency of stock enhancement programs for the sea urchin Hemicentrotus pulcherrimus, information on food algal species, which affect growth and gonad production greatly, is necessary. Since it is difficult to identify species from the macroalgal fragments within the gut contents of the sea urchin by microscopic observation, we tried to apply a DNA barcoding method for gut contents analysis. We used a partial rbcL gene sequence for taxonomic section and newly designed primer sets, respectively, for brown algae and for red algae. Direct sequencing of the PCR products was carried out. Species identification was based on the phylogenetic relationship. We could objectively identify four species and two taxonomic groups (genus or family) in brown algae, and two species and four taxonomic groups in red algae from the gut contents. Sargassum hemiphyllum was the most abundant brown alga in the gut contents but was not dominant in the study site. The result showed the importance of identification to the species level. In addition, red algal epiphytes were detected with brown algal fragments. The DNA barcoding method will enable the researchers to verify the important role of epiphytes as a potential food source.
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References
Vadas RL Sr, Beal B, Dowling T, Fegley JC (2000) Experimental field tests of natural algal diets on gonad index and quality in the green sea urchin, Strongylocentrotus droebachiensis: a case for rapid summer production in post-spawned animals. Aquaculture 182:115–135
Fuji A (1967) Ecological studies on the growth and food consumption of Japanese common littoral sea urchin, Strongylocentrotus intermedius (A. Agassiz). Mem Fac Fish Hokkaido Univ 15:83–160
Endo H, Nakabayashi N, Agatsuma Y, Taniguchi K (2007) Food of the sea urchins Strongylocentrotus nudus and Hemicentrotus pulcherrimus associated with vertical distributions in fucoid beds and crustose coralline flats in northern Honshu, Japan. MEPS 352:125–135
Agatsuma Y, Yamada H, Taniguchi K (2006) Distribution of the sea urchin Hemicentrotus pulcherrimus along a shallow bathymetric gradient in Onagawa Bay in northern Honshu, Japan. J Shellfish Res 25:1027–1036
Ziegler A, Schroder L, Ogurreck M, Faber C, Stach T (2012) Evolution of a novel muscle design in sea urchins (Echinodermata: Echinoidea). PLoS One 7:e37520
Gorokhova E (2006) Molecular identification of the invasive cladoceran Cercopagis pengoi (Cladocera: Onychopoda) in stomachs of predators. Limnol Oceanogr Methods 4:1–6
Blankenship LE, Yayanos AA (2005) Universal primers and PCR of gut contents to study marine invertebrate diets. Mol Ecol 14:891–899
Deagle B, Jarman S, Pemberton D, Gales N (2005) Genetic screening for prey in the gut contents from a giant squid (Architeuthis sp.). J Hered 96:417–423
Steinke D, Hanner R (2011) The FISH-BOL collaborators’ protocol. Mitochondrial DNA 22:10–14
Hebert PD, Stoeckle MY, Zemlak TS, Francis CM (2004) Identification of birds through DNA barcodes. PLoS Biol 2:e312
Hebert PD, Penton EH, Burns JM, Janzen DH, Hallwachs W (2004) Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. PNAS 101:14812–14817
Saunders GW, McDevit DC (2012) Methods for DNA barcoding photosynthetic protists emphasizing the macroalgae and diatoms. In: Kress WJ, Erickson DL (eds) DNA Barcodes. Springer, New York, pp 207–222
Cho SM, Lee SM, Ko YD, Mattio L, Boo SM (2012) Molecular systematic reassessment of Sargassum (Fucales, Phaeophyceae) in Korea using four gene regions. Bot Mar 55:473–484
Agatsuma Y (2013) Hemicentrotus pulcherrimus, Pseudocentrotus depressus, and Heliocidaris crassispina. In: Lawrence JM (ed) Sea urchins: biology and ecology. Academic Press, United Kingdom, pp 461–473
Ishiwatari N, Fushimi H, Maekawa M, Nanba T (1977) Sudies concerning the fishery biology of the sea urchin, Hemicentrotus pulcherrimus (A. Agassiz) in Kaji. Fukui Prefecture -I. La mer 15:146–150 (in Japanese with English abstract)
Taki J, Higashida I (1964) Investigation and problem on introduction of rocks to fishing grounds to enhance the sea urchin Hemicentrotus pulcherrimus in Fukui Prefecture. Aquacult Sci 12:37–47
Goff LJ, Moon DA (1993) PCR amplification of nuclear and plastid genes from algal herbarium specimens and algal spores. J Phycol 29:381–384
Hayakawa Y, Ogawa T, Yoshikawa S, Ohki K, Kamiya M (2012) Genetic and ecophysiological diversity of Cladophora (Cladophorales, Ulvophyceae) in various salinity regimes. Phycol Res 60:86–97
Miyagawa T, Tanaka J, Nagumo T (2007) Seaweed and seagrass flora in Wakasa Bay. Bull Nippon Dent Univ Gener Educ 36:43–50 (in Japanese with English abstract)
Kajimura M (1979) Note on the marine algal flora in the middle part of the Japan sea coast of Honshu (II): Rhodophyta. Mem Fac Sci Shimane Univ 13:97–120
Kajimura M (1978) Note on the marine algal flora in the middle part of the Japan sea coast of Honshu (I): Cyanophyta, Chlorophyta, Phaeophyta. Mem Fac Sci Shimane Univ 12:91–115
Yatsuya K, Nishigaki T, Shirafuji N, Takeno K (2008) Monitoring survey of the seaweed community at Muso-Otani, western Wakasa Bay, Sea of Japan. Bull Kyoto Inst Ocean Fish Sci:27–30 (in Japanese with English abstract)
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular Evolutionary Genetics Analysis Version 6.0. Mol Biol Evol 30:2725–2729
Asahida T, Yamashita Y, Kobayashi T (1997) Identification of consumed stone flounder, Kareius bicoloratus (Basilewsky), from the stomach contents of sand shrimp, Crangon affinis (De Haan) using mitochondrial DNA analysis. J Exp Mar Biol Ecol 217:153–163
Garbary DJ, Clarke B (2001) Apoptosis in trichoblast development in Polysiphonia harveyi (Rhodophyta). Phycologia 40:324–329
Kakuda N, Terao Y, Nakamura T, Inoue T (1970) Growth and feeding habit of the sea urchin, Hemicentrotus pulcherrimus (A. Agassiz) in the laboratory. Aquacult Sci 17:155–165 (in Japanese)
Yatsuya K, Nishigaki T, Itani M, Wada Y (2005) Investigations on drifting seaweed with respect to their mass of occurrence and specific density. Kaiyo monthly 37:516–521 (in Japanese)
Hirata T, Tanaka J, Iwami T, Ohmi T, Dazai A, Aoki M, Ueda H, Tsuchiya Y, Sato T, Yokohama Y (2003) Ecological studies on the community of drifting seaweeds in the south-eastern coastal waters of Izu Peninsula, central Japan. II: seasonal changes in plants showing maximum stipe length in drifting seaweed communities. Phycol Res 51:186–191
Hirata T, Tanaka J, Iwami T, Ohmi T, Dazai A, Aoki M, Ueda H, Tsuchiya Y, Sato T, Yokohama Y (2001) Ecological studies on the community of drifting seaweeds in the south-eastern coastal waters of Izu Peninsula, central Japan. I: seasonal changes of plants in species composition, appearance, number of species and size. Phycol Res 49:215–229
Yatsuya K, Nishigaki T, Douke A, Wada Y (2005) Species composition of drifting weaweed collected in western Wakasa Bay, Japan Sea. Bull Kyoto Inst Ocean Fish Sci:13–18 (in Japanese with English abstract)
Nakajima N, Sugimoto N, Ohki K, Kamiya M (2016) Diversity of phlorotannin profiles among sargassasacean species affecting variation and abundance of epiphytes. Eur J Phycol 51:307–316
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We thank the Wakasamikata Fisheries Cooperative for helping and cooperation with our sampling.
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Nakano, D., Kamiya, M. & Tominaga, O. Molecular identification of macroalgal fragments in gut contents of the sea urchin Hemicentrotus pulcherrimus . Fish Sci 83, 425–432 (2017). https://doi.org/10.1007/s12562-017-1080-6
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DOI: https://doi.org/10.1007/s12562-017-1080-6