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First Record of the Genus Phyllorhiza Spp.../ 22-34 22 First record of the genus Phyllorhiza spp.../ 22-34 First record of the genus Phyllorhiza spp. (Cnidaria; Scyphozoa, Mastigiidae) in the coastal waters of Malaysia Elham Maghsoudlou1, 3, *, Khairun Yahya1, 2 , SimYee Kwang1 , and Mohd Nor Siti Azizah1, 2 1 Centre for Marine and Coastal Studies (CEMACS), Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia 2 School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia 3Department of Biotechnology, Baharan Institute Gorgan, IRAN Received: 2016-09-06 Accepted: 2016-10-08 Abstract This study employed two mitochondrial markers, 16S rRNA and cytochrome c oxidase I (COI) for species identification and resolved the taxonomic and systematic as well as population genetics of Phyllorhiza spp. in the coastal waters of Malaysia. A total number of 65 individuals sampled from three locations representing the three regions fringing the coastline. Phylogenetic trees constructed using Mega 5.0 soft- ware and Kimura 2-parameter with 1000 bootstrap replication were used to test reliability. The molecular data in complement with morphological differentiation revealed, at least three different species for the genus Phyllorhiza, Phyllorhiza punctata inhabit the coasts of Peninsular Malaysia while Phyllorhiza sp.2 and Phyllorhiza sp.3 which were only observed in East Peninsular Malaysia. The results of the current study also highlighted that colour alone is not sufficient for species differentiation of Phyllorhiza spp. The blue morphotype of Phyllorhiza sp.1 observed in East Peninsular Malaysia was genetically identical to the brown morphotype observed in Northwest of Peninsular and recognised as P. punctata while the presence of Phyllorhiza sp.2 and Phyllorhiza sp.3 have never been recorded before in Malaysia. Consequently, under- standing the taxonomy of local and invasive jellyfishes are necessary requirements for their management. Key words: Phyllorhiza spp.; Mitochondrial markers; Jellyfish; Malaysia. 1. Introduction environmental harm to human health. Actually, any species removed from its native range has the po- Every year new invasive animals are reported in the tential to become invasive due to within its normal marine ecosystems (Bax et al., 2003). Invasive spe- range predation, disease, parasite, competition, and cies are generally defined as foreign species whose other natural controls acts to keep population lev- introduction does or likely to cause economic or els in check. If released from this controls, species * Corresponding Author: [email protected] Research in Marine Sciences 23 abundance have the potential to reach levels that in- pass; detailed morphology (Mayer, 1910; Kramp, terfere with or displace local fauna such effect may 1961), reproduction and survival (Rippingale and occur immediately, depends on the characteristics Kelly, 1995), behaviour and pray capture (D’am- of the individual species and the conditions into bra et al., 2001), and the population dynamics and which it is introduced it may be never realized at all production of P. punctata in Laguna Joyuda (Joyu- or accrued after some period of delay (Ray, 2005b). da and Rico, 1990). P. punctata has large size with Phytoplankton has an important link in the food a characteristic colouration and inhabits the upper chain; they are agent for primary production and layer of the water. The living specimens are brown through the process of photosynthesis they abtin with obvious white spots and have eight radi- energy. An invasive species can lead to a disappear- al canals, the bell diameter varies between 2-470 ance and reduction in phytoplankton availability mm and the weight ranged between 0.26-3882.2g and consequently affect a marine ecosystem food (Haddad and Nogueira Júnior, 2006). Sim (2009) web resulting in reduced zooplankton and fish and Chuah (2012) reported its occurrence in Malay- abundance and changed feeding habits of resident sia based on morphological traits. fish (Ray, 2005a). Phyllorhiza is an efficient invasive organism Phyllorhiza punctata Von Lendenfeld, 1884 is a that caused immense damage in marine ecosys- well known invasive Scyphomedusae belonging to tem. Very limited genetic information is available the order of Rhizostomeae and family of Mastigii- for the taxonomy and classification of this genus dae and indigenous to the tropical western Pacific and to date, no published data recorded regarding and have successfully migrated globally (Graham the molecular taxonomy of this important species et al., 2003; Ray, 2005a). Prior to 1950, this species in Malaysia. There has been complication in dis- was the only known in the Indo-Pacific Ocean. To- tinguishing between Phylloriza and another genus, day it has been documented in many locations of Mastigias which also a member of the family Mas- the world wide such as Gulf of Mexico (Graham tigiidea. Stiasny (1924) first reported the genetic et al., 2003; Chilton et al., 2011), Southern Aegean description of these two genera, although some coast of Turkey (Gülşahin and Tarkan, 2012), Ital- confusion had remained. Similarly, another species, ian waters, west of Mediterranean Sea (Boero et al., M. albipunctatus identified from Jamaican waters 2009), and the coastal waters of Brazil (Haddad and (Vannucci, 1964) was also regarded a likely mis- Nogueira Júnior, 2006). ident. Bayha and Graham (2011) confirmed the Phyllorhiza punctata prey upon many econom- presence of the Mastigias in the Atlantic basin and ically important organisms, including crustaceans, suggested that the origin of this genus may be in the bivalve larvae, and fish eggs and larvae, within central Indo-Malayan region. According to Dawson their native range resulting in negative effects on and Jacobs (2001) molecular genetics techniques the economy and ecology of the marine ecosystems can facilitate determination of species boundaries (Chilton et al., 2011). For example, Graham et al. as well as correcting morphological misidentifica- (2003) reported, indirect evidence that the presence tions. Thus, its management is critical and the in- of P. punctata in the Gulf of Mexico decreased the vestigation of the taxonomy and genetic population total catch of white shrimp (Litopenaeus setiferus) of the genus Phyllorhiza is a prerequisite for formu- in Mobile Bay, Alabama and Mississippi Sounds by lating strategies to achieve this. The data would be 27% (clogging of nets, damage to boat intake and an important contribution towards the management fishing gear and area closures). of this group, which is fast becoming an unwel- Several aspects of this invasive species have come and sometimes deadly feature, in many global been investigated in the literature. These encom- waters in the past recent years. The current study 24 First record of the genus Phyllorhiza spp.../ 22-34 discerns the characters of genetic diversity, phylo- Island, 2. Langkawi Island in (Northwest), and 3. genetic and phylogeography among the populations Kuala Terengganu (East) (with all three located in inhabiting the Peninsular Malaysian waters. Peninsular Malaysia) from November 2010 to Sep- tember 2011 (Figure 1, Table 1). Two methods were 2. Materialsfrom the surface and andmethods towing methods in deeper watersused forspecimens. sampling; The the specimens push net (scoop from Kualanets) for the specimens from the surface and towing methods Terengganu obtained from fishermen. Seven morphotypes of Phylloriza spp. collected in deeper waters specimens. The specimens from from three populations: 1. Muka Head, Penang Kuala Terengganu obtained from fishermen. 20 cm 18 cm 17 cm A) Phyllorhiza sp.1 in Langkawi B) Phyllorhiza sp.1 in Muka C) Phyllorhiza sp.1 in Head Terengganu 15 cm 15 cm 13 cm D) Pink morph of Phyllorhiza sp.2 E) Blue morph of Phyllorhiza F) Purple morph of in Terengganu sp.2 in Terengganu Phyllorhiza sp.2 in Terengganu 15 cm G) Brown morph of Phyllorhiza sp.3 in Terengganu Figure 1. Different morphotype of Phyllorhiza spp. (A-G) 4 Research in Marine Sciences 25 Table 1. Sampling site, tag label, number of morphological types, sample size, sampling date and GPS coordinates of Phyllorhiza spp. populations investigated along the Malaysian coast Location and No. of Morphological Sampling Region Location Coordinate Tag Types and samples Date 1 Penang (MH) Northwest 1 20 Nov 2010 5°24′00″N100°14′20″E Peninsular Langkawi 2 Northwest 1 20 Nov 2010 6°21′N 99°48′E (L) Peninsular 3 Terengganu (T) East 5 25 Sep 2011 5°20′0″N103°9′0″E Peninsular These specimens identified as Phylloriza ac- saicus, Kayal (2012- JN700937.1, JN700940.1from cordingThese to Mayer specimens (1910) identifiedand Kramp as (1961) Phylloriza key, accordingUSA) Cassiopea to Mayer sp. (1910)Bridge (1995-and Kramp U19374.1 (1961) from key, but even within each presumed species colour vari- USA) and Nemopilema nomurai Zhang (2008- but even within each presumed species colour variations occurred. Total genomic DNA of 65 ations occurred. Total genomic DNA of 65 speci- EU373727.1 from China) used as a outgroups for mens werespecimens extracted were using extracted AQUAGENOMIC™ using AQUAGENOMIC™ kit comparisons kit in (Multigeographic Target distance Pharmaceuticals, analysis in 16S Salt (Multi TargetLake City,Pharmaceuticals, Utah 84116), Salt following Lake City, theUtah manufacturer rRNA gene.s’ protocol.Unfortunately, The targetamplification genes, 16S of Phyl rRNA- 84116), following the manufacturers’ protocol. The lorhiza spp. for COI gene was unsuccessful even target genes,was amplified 16S rRNA using was
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