Distribution of the Northern Star Coral Astrangia poculata

Distribution of the Northern Star Coral (Astrangia poculata) and its Symbiotic Zooxanthellae on New Jersey Artificial Reefs. Peter F. Straub, Professor of Biology, DSO Tara Harmer, Assistant Professor of Biology School of Natural Sciences & Math Richard Stockton College of NJ

Collaboration-Undergraduate Research and Teaching Links n n n Stockton Scientific Diving Program- Straub -DSO BIOL 2175 - Scientific Diving-Straub Fall 07 BIO 4211 - Molecular Evolution- Harmer Fall 07 BIOL 4215 -Biotechnology. Straub, Spring 08 BIOL 4800 - Biology Research- Harmer Fall/Spring 07/08 BIOL 2175 Fall 2007

Astrangia poculata (syn. A. danae) -Northern Star Coral http: //www 2. aims. gov. au/coralsearch/html/ n Cnidaria › Anthozoa › Hexacorallia › Scleractinia › Faviina › Rhizangiidae › Astrangia n Found from the sub-tropics to the temperate zone <5 C. Most corals live in tropical to sub-tropical waters above 20 C and cannot tolerate high turbidity n Colonies can be encrusting, massive (mounding) or branching n The individual coral skeletons or corallites are well defined, circular, compact, and up to 10 mm in diameter n Living coral tissue may be translucent to dark brown depending on the presence and number of zooxanthellae n May contain zooxanthellate, or not, given local light conditions.

Hypothesis n Since the distribution of northern star coral in NJ is patchy, only found on hard surfaces such as wrecks, is there any detectable genetic differentiation among local collection sites and between disjunct populations.

Methods n n n n Site selection Collect corals by scuba Extract chromosomal DNA Amplify ss. RNA (18 S) sequence by PCR DNA Sequence Align sequences (Vector n. Ti) Look for relatedness in coral pops (Mac. Clade and PHYLIP)

NJ Coral Collection sites 7 sites, 15 dives

Lemuel Burrows, torpedoed, 3/14/1942, 80 ft water Car float, 60 ft John Marvin Clam Dredge Sunk 1/16/1992 70 ft. Jet Trader, yard oiler, 95 ft. , artificial reef, sunk 9/2/05 http: //njscuba. net/sites

Wreck of the Almirante (the flour wreck) off Atlantic City NJ (approx 8 mi) at 70 ft. United Fruit Co. vessel (378 ft x 50 ft) sunk in collision Sept 6, 1918 w/ Navy tanker USS Hisko, later wire dragged 2 x, depth charges, fairly broken up http: //njscuba. net Coral collections 8/25/07, 8/29/07 and 10/6/07 Underwater photos David Roche

MV Atlantus Charters Sea Girt, Clam Dredge, Depth 70 ft sunk 8/1990. Coral Collection- 7/28/07 Northern Star Coral Astrangia poculata Photos Pete Straub

Almirante (Flour)

Dave Pete Mike Almirante (Flour)

Almirante

Coral plus mussels Coral heads Photos P. Straub & T. Harmer Coral heads Coral plus anemone

Individual coral polyps Kept 10 gallon aquaria, filtered, Feed zooplankton, commercial zooplankton (preserved). Coral polyps extended Photos P. Straub & T. Harmer

DNA Extractions Remove tissue from single corallite using scalpel blade tip (like cutting grapefruit) Qiagen DNAeasy (column method) Kits Eluted DNA from columns with Sterile water Measure DNA content at A 260 with spectrophotometer Analyze by agarose gel electrophoresis- High MW Adjust concentration for PCR reaction

Polymerase chain reaction (PCR) w/universal eukaryotic small subunit (ss)RNA- (18 S) 1. PCR Reaction Conditions 100 ng chromosomal DNA Qiagen TAQ polymerase Quiagen coraload buffer 0. 2 u. M d. NTP’s Forward Primer NS 1* Reverse Primer Euk 516 R** 1 cycle 35 cycles 1 cycle 94 94 52 72 72 C, C, C, ~550 bp 1: 00 min 1: 30 min 2: 00 min 10: 00 min 2. Analyze by agarose gel electrophoresis 3. Purify PCR products using Promega SV gel and PCR product cleanup colums kit *White et al. , 1991 **Di’ez et al. , 2001

Cloning amplified products from Coral PCR • Purified coral PCR products were cloned using a p. Gem. T cloning kit (Promega kit) simplifies cloning as PCR products may have an “A” base overhang and is simple to ligate to p. Gem. T plasmid vector. • Ligation products were transformed into E. coli. • E. coli colonies were selected on ampicillin. • DNA plasmid minipreps were performed on individual E. coli cultures • Plasmids concentration measured by spectrophotometer A 260

Automated Fluorescent DNA Sequencing • Use ~200 ng coral plasmid DNA, Beckman Coulter DCTS master mix With forward (T 7) and reverse (sp 6) sequencing primers • Separation Beckman Coulter CEQ 8000 Genetic Analyzer • Export sequence trim out plasmid vector sequences