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Bivalvia: Ostreidae): a New Species in Zhanjiang, China

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Bivalvia: Ostreidae): a New Species in Zhanjiang, China Aquat. Living Resour. 26, 273–280273-280 (2013) Aquatic ©c EDP Sciences, IFREMER, IRD 2013 DOI: 10.105110.1051/alr/2013065/alr/2013065 Living www.alr-journal.org Resources Mitochondrial DNADNA and morphologicalmorphological identificationidentification of CrassostreaCrassostrea zhanjiangensis sp. nov.nov. (Bivalvia: Ostreidae): a new species in Zhanjiang, China Xiangyun Wu,ShuX, Shu XiaoIAO and Ziniu Yua CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese AcademAcademyy of Sciences, Guangzhou 510301, P.R.P.R. China Received 9 July 2013; Accepted 25 September 2013 Abstract –- Cupped oysters (Ostreidae, genus CrassostreaCrassostrea)) occupy nearshore marine and estuarine habitats world-world­ wide, providing many ecosystem goods and services as well as being a commercially important group of bivalves. In this study, the species identificationidentification of an “adulterant”“adulterant” oyster with small body size, which is often misidentifiedmisidentified as a “young individual”individual” of other sympatric species, including C. sikamea and C. hongkongensishongkongensis,, was determined for the firstfirst time, based on molecular markers (partial mitochondrial cox1coxi and rrnL genes), phylogenetic analysis, and mor mor-­ phometric approaches. This novel species, C. zhanjiangensiszhanjiangensis,, commonly known as the “cat“cat ear oyster”oyster” in Guandu (a famous estuarine oyster farming region of Zhanjiang, Guangdong Province, China), appears to potentially influenceinfluence the eefficiencyfficiency of C. hongkongensis spat collection due to niche competition on spat collection devices. Phylogenetic analyses confirmconfirm its status as the most basal taxon of the Indo-PacificIndo-Pacific Crassostrea. A comparative study of the shell characteristics of C. zhanjiangensis ,, and other Crassostrea species revealed several distinctive morphological traits, including a generally smaller body size, a deeply cupped left valve, and a right valve that is convex in adults but flatflat in young individuals. Other distinctive features of the new species include life cycle traits that are unique compared with the sympatric C. hongkongensis and C. sikamea species, such as a higher growth rate in the fast growth phase after settlement, followed by a significantlysignificantly slower growth rate and mass mortality during subsequent life stages. This study provides the basic information necessary for further ecological and population genetic studies on this new species. Keywords: Mitochondrial gene / PCR / Molecular identificationidentification / Phylogeny / New species / Oyster / crassostreaCrassostrea 1 Introduction Fujian, to south in the Guangdong-Guangxi provinces these are: C. gigas (the PacificPacific oyster), C. angulata (the Portuguese Oysters (Family Ostreidae) occupy nearshore marine and oyster) and C. hongkongensis (the Hong Kong oyster) (Xia estuarine habitats in temperate to tropical latitudes worldwide 2008;; Xia et al. 2009;Wuetal.; Wu et al. 2010;Lietal.; Li et al. 2013). In recent (Ruesink et al. 2005). Being ecosystem engineers, they can years, increasing pressure from local markets in Hong Kong, provide many ecosystem goods and services by creating habi- Macau, Taiwan, and mainland CChinahina have prompted expan-expan­ tat then used by other species, and can modify the physi- sion in the scale of oyster farming in China. Although oyster cal and chemical environment, with major consequences for hatcheries are widely used for seed production, spat collection estuarine populations, communities, and food webs (Jackson from the wild is still extensively practiced for Hong Kong and et al. 2001; Ruesink et al. 2006). Oysters of the genus Cras- Portuguese oysters. Spat collection is economical, but both the sostrea are a widespread and commercially important group quality and quantity of C. hongkongensis spat, from which is of bivalves. The coastal areas along the South China Sea are grown the most popularly cultured oyster in southern China, species-rich regions of Crassostrea oysters Their cultivation are aaffectedffected by the variable flowflow of coastal rivers. Furthermore, is reputed to have several hundred years’ history (Lam and oyster spat collected from didifferentfferent areas exhibit didifferencesfferences in Morton 2003) and is now becoming one of the largest coastal production traits such as growth rate, meat flavour,flavour, and even industries, with recent annual landings of around 3.5 million disease resistance. Several areas still produce good quality tons (nearly 78% of total world production) (Xia et al. 2009). oyster seed, including Huian and Putian in Fujian Province for Three oyster species dominate oyster aquaculture in the coastal C. angulataangulata,, and the Hengqin-Sanzhao area (Zhuhai), Beijin waters of China; in order from north in Liaoning-Shandong, Harbour (Yangjiang) and Guandu (Zhanjiang) in Guangdong Province for the Hong Kong oyster (Xiao and Yu 2008;Li; Li a Corresponding author: [email protected] et al. 2013). SpatSpat collected in these aareasreas is usually cultured Article published by EDP Sciences Downloaded from https://www.cambridge.org/core. IP address: 193.191.134.1, on 04 Aug 2017 at 13:03:46, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1051/alr/2013065 274 X. Wu et al.: Aquat. Living Resour. 26, 273–280 (2013) Table 1. Species identificationidentification of Crassostrea oysters collected from oyster farms in didifferentfferent locations and at indicated dates. Location Date Number of individuals Species (number;(number; percentage) Guandu, Zhanjiang November, 2011 128 C. hongkongensis (76; 59%) C. zhanjiangensis (43; 34%) C. sikamea (9; 7%) Donghai Island, Zhanjiang December, 2011 84 C. hongkongensis (46; 55%) C. angulata (38; 45%) Guandu, Zhanjiang May, 2012 187 C. hongkongensis (83; 44%) C. zhanjiangensis (89; 48%) C. sikamea (15; 8%) Henqing, ZhuhaiAugust, 2012 66 C. hongkongensis (66; 100%) locally, but is also sold for cultivation elsewhere. Thus, eeffiffi-­ (10 ×x 1010 ×x 11.5.5 cm) or oyster shells and transported to the ciency of spat collection can potentially aaffectffect not only local laboratory alive. oyster cultivation but also the oyster industry along the coast of the South China Sea. The most important factor influencinginfluencing the eefficiencyfficiency of spat collection is niche competition on spat 2.2 DNA extraction, extraction, PCR amplification amplification and and sequencing sequencing collection devices (e.g., concrete tiles) by sympatric species. This study was initiated following a series of fieldfield inves-inves­ Whole genomic and mitochondrial DNA of each indi-indi­ tigations of the Hong Kong oyster in the farming regions of vidual was extracted from adductor muscle or visceral mass Zhanjiang, Guangdong Province (Li et al. 2013). Our orig-orig­ using the TIANamp Marine Animals DNA kit (Tiangen, inal intention was to collect specimens of cultured young Beijing). A partial cox1 segment was amplifiedamplified by polymerase C. hongkongensis individuals for population genetic analysis. chain reaction (PCR) with primer pairs of COIL1490 (5(5'-- As the shell morphology of this species is phenoplastic and GGTCAACAAATCATAAAGATATTGG-3GGTCAACAAATCATAAAGATATTGG-3')) and COIH2198 greatly aaffectedffected by habitat, we employed partial mitochondrial (5(5'-TAAACTTCAGGGTGACCAAAAAATCA-3').-TAAACTTCAGGGTGACCAAAAAATCA-3 ). The The cox1 gene sequencing as a molecular marker for species identi-identi­ primer pair of 16sar16sar (5(5'-CGCCTGTTTATCAAAAACAT-3')-CGCCTGTTTATCAAAAACAT-3 ) fication.fication. Unexpectedly, a very high proportion of the randomly and 16sbr16sbr (5(5'-CCGGTCTGAACTCAGA-CCGGTCTGAACTCAGA TCACGT-3TCACGT-3'))was was sampled specimens was found to represent oyster species other used to amplify the partial rrnL segment. PCR reactions than C. hongkongensis.. According to descriptions from expe-expe­ were performed in a 25 //lµl volume with 0.5 //lµl template DNA rienced local oyster farmers, most non-Hong Kong oysters are (approximately 30 ng), 0.5 /lµl10mMdNTPmix,2.5 10 mM dNTP mix, 2.5 /lµl + considered to be C. sikamea (the Kumamoto oyster), a species 1010 ×x bufferbuffer (Mg22+ plus), 11 /lµl of each primer (10 /M),µM), and sympatric with C. hongkongensis and commonly considered 0.25 /lµl(1U)Ex (1 U) ExTaqTaq polymerase (Takara, Dalian, China). The as an “adulterant“adulterant oyster”oyster” in Hong Kong oyster aquaculture. PCR reactions were performed on an ABI Veriti thermal cycler Our initial molecular data supported this claim; however, fur-fur­ (Applied Biosystems, California, USA) with the following ◦ ther studies unexpectedly identifiedidentified some cox1 sequences that parameters: pre-denaturation at 94 °CC for 11 min, followed by ◦ ◦ match neither those of C. sikameasikamea,, nor those of any other 35 cycles of 94 °CC for 20 sec, 45-55 45−55 °CC annealing temperature ◦ ◦ known species of the genus Crassostrea.. Therefore, it appears (52(52° for cox1 and 50 °CCfor for rrnLrrnL)) for 20 sec, extension at ◦ ◦ these individuals represent a hitherto undescribed species from 72 °CC for 11 min, and a finalfinal extension step at 72 °CCfor5min. for 5 min. this region. PCR products were separated by electrophoresis on a 1%1% The objective of this study was to determine the specificspecific agarose gel, purified purified with QIAquick PCR PurificationPurification kit identity of this Zhanjiang
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