60 



Fishery Bulletin 92(1), 1994 



loci: ACP-2* (acid phosphatase); ADA* (adenosine 

 deaminase); ADH* (alcohol dehydrogenase); sAAT-1* 

 (aspartate aminotransferase); EST-1* (esterase); 

 GPI-B* (glucose phosphate isomerase); and PEPB' , 

 PEPD * , and PEPS' (peptidases). Techniques for ver- 

 tical starch gel electrophoresis, details of grinding 

 and running buffers, starch composition of gels, 

 protein staining, and interpretation of banding pat- 

 terns may be found in Bohlmeyer (1989) and 

 Bohlmeyer and Gold (1991). Designation of allelic 

 variants was based on relative mobility to the most 

 common allele (Allele * 100). 



All individuals collected (109 total) were assayed 

 for 104 mtDNA restriction sites with 13 restriction 

 enzymes: BamKl, Bell, EcoRV, Hindlll, Ncol, Nsil, 

 PstI, Pvull, Seal, Spel, Stul,Xbal, and Xmnl. Meth- 

 ods used to assay mtDNAs of individual fish may 

 be found in Gold and Richardson (1991). Homology 

 of fragments from single digestions was tested by 

 multiple, side-by-side comparisons. Variant patterns 

 exhibiting only a single band of greater than 15 kb 

 were tested for homology by using double digestions 

 with BawHI as described in Gold and Richardson 

 (1991). 



Red drum from Mosquito Lagoon were initially 

 subdivided into year classes and tested for hetero- 

 geneity in both allozyme and mtDNA haplotype fre- 

 quencies. Year classes (number of individuals) were 

 1985 (17), 1986 (25), 1987 (11), 1988 (7), and 1989 

 (49). No significant heterogeneity (P>0.05) in 

 allozyme or mtDNA haplotype frequencies was 

 found among year classes. Subsequent data analy- 

 ses employed three test groups: 1) red drum from 

 Mosquito Lagoon; 2) red drum from the northeast- 

 ern Gulf; and 3) red drum from the Carolina coast. 

 Data for the latter two were taken from Gold et al. 

 (1993, 1994) and represent red drum from the fol- 

 lowing localities: northeastern Gulf — Apalachicola 

 Bay, Riviera Bay, and Sarasota Bay (west coast of 

 Florida); and Carolina coast — Calibogue Sound, 

 Charleston Bay, and North Inlet (South Carolina), 

 and the Pamlico River and Oregon Inlet (North 

 Carolina). A map showing these localities may be 

 found in Bohlmeyer and Gold ( 1991 ). A summary of 

 allele frequencies at the nine polymorphic allozyme 

 loci and the distribution of mtDNA haplotypes in 

 each test group are given in Appendix Tables 1 and 

 2, respectively. 



For allozyme data, tests of Hardy-Weinberg equi- 

 librium expectations and generation of Nei's (1978) 

 unbiased genetic distance were accomplished by 

 using BIOSYS-1 (Swofford and Selander, 1981). 

 Deviations from Hardy-Weinberg expectations were 

 tested by using pooled genotypes and the chi-square 

 statistic with one degree of freedom. Significance 



testing of allele-frequency differences among test 

 groups was accomplished by using 1) the G-statis- 

 tic (Sokal and Rohlf, 1969) on contingency tables of 

 allele counts and the BIOM-PC program (Rohlf, 

 1983), and 2) the ^-statistic (DeSalle et al., 1987) 

 on arcsin, square-root transformed allele frequen- 

 cies. For mtDNA data, significance testing of 

 mtDNA-haplotype frequency differences was carried 

 out by using the G- and V-statistics as described 

 above and a Monte Carlo randomization procedure 

 (Roff and Bentzen, 1989). Nucleon diversities and 

 intra- and inter-populational nucleotide sequence 

 diversities were estimated by using equations in Nei 

 and Tajima (1981). Analysis of mtDNA data was 

 facilitated by the Restriction Enzyme Analysis Pack- 

 age (REAP) of McElroy et al. (1992). Significance 

 levels for multiple tests performed simultaneously 

 were adjusted after Cooper (1968). 



Results 



No significant deviations from Hardy Weinberg equi- 

 librium expectations at any of the nine polymorphic 

 allozyme loci were found following corrections for 

 multiple tests. Two significant deviations were found 

 in uncorrected tests: at GPI-B* (P=0.015) and PEPS' 

 (P=0.012) in the northeastern Gulf. Both deviations 

 appeared to be due to rare homozygotes for low fre- 

 quency alleles. One new allele (Allele * 110 at EST- 

 l') was found among Mosquito Lagoon fish at a fre- 

 quency of 1.2 percent (Appendix Table 1). 



Estimates of allozyme variation (Table 1) indicate 

 that red drum from Mosquito Lagoon have fewer 



