Journal of Mosquito Research, 2013, Vol.3, No.8, 58-64
ISSN 1927-646X
http://jmr.sophiapublisher.com
60
Table 1 Single digested fragments of
COI
gene of
Cx. quinquefasciatus, Ae. aegypti, Ae. albopictus, An. stephensi
with
Dra
I,
Taq
I,
Mbo
I,
Msp
I,
Bgl
II
and
Eco
RI
Table 2 Double digested products of
COI
gene of
Cx.
quinquefasciatus
,
Ae. aegypti
,
Ae. albopictus
and
An. stephensi
with a combination of
Mbo
I
+ Dra
I
and
Mbo
I
+ Taq
I enzymes
genetic polymorphism among the various mosquito
species was evaluated using the POPGENE 1.31
version. A total of 8 bands or loci representing all the
four mosquito species were considered (Figure 1). The
percent polymorphism obtained was 87.5% among the
different mosquito species. The genetic diversity
among the species indicated by the Shannon Index
and the gene diversity index of 0.34 each as shown in
Table 3. The minimum genetic distance was obtained
between
Cx. quinquefasciatus
and
Ae. albopictus
of
0.28,
whereas the maximum genetic identity was also
observed as 0.75 between these two species (Table 4).
Consensus tree generated by using the data showed
that
Aedes
species (
Ae. aegypti
and
Ae. albopictus
)
and
Cx. quinquefasciatus
are closely placed (Figure 2).
On the other hand
An. stephensi
is represented in the
separate branch with maximum distance of 0.69 with
the other three mosquito species and having least
identity.
Figure 1. Double digestion with
Mbo
I +
Dra
I restriction
enzymes
Note: Lane 1 & 6: 50 bp DNA Ladder; Lane 2-5: MboI+ DraI
digested samples of Cx. quinquefasciatus, Ae. aegypti, Ae.
albopictus and An. stephensi; Lane 7: Blank
3.
Discussion
The first evolutionary tree of Culicidae was
constructed by Ross (Ross, 1951) based on an
intuitive interpretation of comparative bionomics and
morphological data. The tree reflected the traditional
division of family Culicidae into three subfamilies:
Anophelinae, the basal lineage; Toxorhynchitinae, the
intermediate lineage; and Culicinae, the large derived
lineage. The relationships illustrated by Ross (Ross,
1951)
are those more or less traditionally accepted and
unchallenged by later workers until Reinert, Harbach
and Kitching (Reinert et al., 2009) examined the
generic relationships and higher classification of the
family based on cladistic analyses of morphological
data. Reidenbach et al., 2009
examined the
phylogenetic relationships of four mosquito species,
one from each of genera
Aedes
,
Anopheles
,
Culex
and
Toxorhynchites
,
using 18S and 5.8S rDNA sequences.
Their analysis placed
Anopheles
in a position basal to
the other species. In a parsimony analysis-
Toxorhynchites
was placed as the sister group of
Aedes
+
Culex
,
whereas
Aedes
was the sister group of
Toxorhynchites
+
Culex
in a maximum likelihood tree.
Mosquito
Dra
?
Taq
?
Mbo
?
Msp
?
Bgl
?
Eco
R
?
Cx. quinquefasciatus
280
bp 300 bp
80
bp 200 bp 300 bp
90
bp 190 bp 290 bp
_
_
_
Ae. aegypti
_
_
120
bp 180 bp 280 bp
180
bp 400 bp _
_
Ae. albopictus
280
bp 300 bp
200
bp 380 bp
120
bp 180 bp 280 bp
260
bp 320 bp _
_
An. stephensi
280
bp 300 bp 280 bp 300 bp
190
bp 390 bp
_
_
_
Mosquito
Mbo
?
+
Taq
?
Mbo
?
+
Dra
?
Cx. quinquefasciatus
90
bp
190
bp
250
bp
90
bp
180
bp
280
bp
Ae. aegypti
110
bp
190
bp
280
bp
120
bp
180
bp
280
bp
Ae. albopictus
90
bp
190
bp
280
bp
120
bp
180
bp
280
bp
An. stephensi
90
bp
190
bp
300
bp
120
bp
180
bp
280
bp