Page 1 - Mullus1991

Comp. Biochem. Physiol. Vol. 99B, No.3, pp. 719-122, !991 0305-0491 /91 SJ.OO + 0.00
Printed in Great Britain ::Q 199 l Pergamon Press pie

BIOCHEMICAL TAXONOMIC DIFFERENTIATION
BETWEEN MULLUS BARBATUS AND
MULLUS SURMULETUS (PISCES, MULLIDAE)

M. CAMMARATA, N. PARRINELLO and M. ARCULEO*
Institute of Zoology, University of Palermo, Via Archirafi 18, 90123 Palermo, ltaly; and
*Institute of Zootechnics. University of Reggio Calabria. P.zza S. Francesco 7, 89061 Gallina (RC), ltaly

(Receired 8 Jcmuary 1991)
Abstract-1. The electrophoretic data from seven en7.ymatic systems, cod ifying for 20 loci, and the
patterns of generai proteins from some tissues of ,),fullus harhcnus and M1dlus surmulelus were examined.
2. The value of the genetic distance index as per Nei is 0.068, this criterium showing a high similarity
j between lhe two species.
3. Species-specific patterns were found for the SOD enzyme and generai protcins of the m uscle.

INTRODUCTION E/ectrophorcsis
Polyacrylamide gel electrophoresis (PAGE) was carried
The two species J'l4ullus barbatus and Mullus surmu-
out as described by Davis ( 1964). The samp!e, 1-5 111 in
letus from the whole Meditcrrancan basin havc 100111 sample-buffer, was deposi led into each well of thc
been investigated with regard to reproduction, ethol- spacer slab gel { 16 x 16 cm, 2 mm thick) and venically run
ogy and growth (Scaccini, 1947; Bougis, 1952; Gharbi al a constant currenl of 40 mA.
and Ktari, 1981; Arculeo et al., 1989). I-Iowever, little After migration, lhc gel slab was stained with Coomassie
information concerning their structure and genetic Brillianl Blue for generai protcin pattern, unless otherwise
indicatcd. Thc rari o frontis (Rrl of cach band was calculaled
variability is at present available (Basaglia and
by u sing bromophenol blue as marker.
Callegarini, 1988; Cammarata el u/., 199 1 ).
A high degree of intraspecifìc variability, particu- Homoge11ate preparation
Jarly enhanced among the juvenilia, as measured
Tissues from liver, heart, musclc, gonads and eye were
thro ug h biometrie parameters, renders classifi.cation homogenized in one volume of NaCI l M at D'C. T he
a difficult task to perform (Castelnuovo, 1936; Bougis, extracts were centrifuged a t 6000 .r: a t 4"' C for 30 mi n alter
1952), even though the two species differ fo r habitat, wbich the supernatant was papcr-filtercd lo remove the lipid
patlern of growth and morphological characters layer. The filtrate was storcd at - 75"C.
(Tortonese, 1975).
Gel swining for isoenzymes
Analysis of genetic variation, as carried out on an
The staining solutions utili7.ed lo visualize each enzymc
adequate number of loci by electrophoresis. is a well-
were as follows:
known practical means of investigating thc genetic
Luctute dehydrogenase (LDH, EC 1. 1.1.14). Sixteen ml
structure of natura] population, as well as of high- 0.1 M pH 7.4 phosphatc buffer (PB): 16 mi lactate buffer
lighting the occurrence of a possible taxonomic (7.37 mi acid DL-laclate 90%, 92.63 mi PB); 48 ml MTT
relationship (Ayala, 1975; Nei, 1978; Graf, 1982). 0.001% wjv in distillcd water; 4.8 mi phenazine methasulfate
The present research aims at assessing. by electro- (PMS) 0.00 l wjv in disti !led water; 80 mg !X-nicotinamide
j phoresis of a few isoen zymes, the genetic distancc adcninc dinucleotide (NAD) (McKenzie et al .• 1983).
Mci/ate dehydrogenase (.MDH, EC 1. 1.1.37). Eighty mi
between J>,;fullus barbarus and i'v!ullus surmuletus (mak-
ing use of Nei's Index, 1978), as well as fi.nding any Tris-HCI 0.1 M p H 8.0: 80 mg aci d D L-malie; 2.4 mg PMS;
12.8 mg MTT; 32 mg NAD (Graf and Meylan, 1980).
possìble diagnostic loci and drawing a comparison
Alcohol dehydroKenase (ADH, E C 1.1.1.1). Eighty ml PB
between the populations examined.
0.1 M pH 7.0; 3.2 mi ethanol; 40 mg NAD; 3.2 mg PMS:
16 mg MTT (Graf and Meylan, 1980).
c:J.-Giycaopho~phate clehydrogenase (GPD, EC 1.1. 1.8).
Eighty mi Tris·-HCI 0.1 M pH 7.1; 160 mg DL-!X -glycero-
MATERIALS ANO i\lETHODS
phosphate disodium salt: 40 mg NAD; 1.6 mg PMS; 16 mg
Seventy M. barbatus specimens and 65 M. surmuletus. MTT; 0.8 mi MgCI 0.2 M (Graf and Meyland, 1980).
from various locations of Sicily (Gulf of Gela, Gulf of Superoxide dismutase (SOD, EC 1. 15.1. 1). Eighty m i Tris-
Castellammare, G ulf of Palermo), of Calabria (Vibo Valentia, HCI 0.5 M pH 8.0; 16.8 mg MTT; 16.8 mg PMS (Philipp
Bovalino), and Egadi Islands were examined and 1he follow- el al., l 979).
ing parameters recorded: standard length (S. L). totallength Sorbito/ dehydrogenase (SDH, EC 1. 1.1.!4). Eighty m i
(f.L.), weight, sex and maturity leve!. M. barbatuJ size Tris-HCl 0.25 M pH 8; l g sorbito!; 12 mg NAD; 5 mg
ranged from 8 to 16 cm (S.L.), 11.83 (SD ± 1.99) being the PMS: 12 mg MTT (Philipp et al .. 1979).
mean value, and i\.f . . mrmuletu.l standard length ranged from Jsocitrme dehydrogena.~e (JDH. EC l.l .l .42). Eighty mi
lO lo 20 cm with a mean of 14.89 (SD ± 2.48). The frozen Tris-HCI 0.05 M pH 8.5: 80 mg DL-isocitric acid ; 16 mg
fish, stacked in suitablc containers, were taken lo the NADP; 1.6 mg PMS; 12.8mg MTT, !.6ml MgSO, 0.1 M
laboratory, and kept there al -20'C, unti! they were used. (Graf and Meyland, l 980).
719