ISSN 20790597, Russian Journal of Genetics: Applied Research, 2011, Vol. 1, No. 5, pp. 390–395. © Pleiades Publishing, Ltd.

, 2011.
Original Russian Text © M.B. Udalov, G.V. Benkovskaya, 2010, published in Ekologicheskaya Genetika, 2010, Vol. 8, No. 3, pp. 61–66.

Change in the Polymorphism Level in Populations

of the Colorado Potato Beetle
M. B. Udalov and G. V. Benkovskaya
Institute of Biochemistry and Genetics, Ufa Research Center, Russian Academy of Sciences, pr. Oktyabrya 71,
Ufa, 450054 Russia
Received December 31, 2009
Accepted for publication July 15, 2010

Abstract—Data on changes in the phenetical structure of populations of the Colorado potato beetle Leptino
tarsa decemlineata Say in the Southern Urals (territory of the Republic of Bashkortostan) are presented. A
decrease in the level of phenetical polymorphism between 1994 and 2002 was registered. The results of labo
ratory experiments give us good grounds to believe that the selective effect of insecticides is the main reason
for the decrease in the level of intrapopulation polymorphism.
Keywords: Colorado potato beetle, Leptinotarsa decemlineata Say, population structure dynamics, population
phenetics, population genetics, polymorphism, Southern Urals
10.1134/S2079059711050157

INTRODUCTION 1985; Eremina, Denisova, 1987; Klimets, 1997;

The history of distribution of the Colorado potato
beetle (Leptinotarsa decemlineata Say) outside of its
native range has been closely related to the timeline of
appearance and spread of new populations resistant to
insecticides (Roslavtseva, 2009; Udalov et al., 2009). It
is conjectured that the rapid expansion of this species,
which can be called ecological explosion (Fasulati,
2002), stems from the high level of its intraspecific poly
morphism and relatively recent origin, which are sup
posedly responsible for its amazing plasticity and adapt
ability (Ushatinskaya, 1981; Fasulati, 2002; Tower,
1906).
It is evident that in order to successfully control
and regulate the abundance of the Colorado potato
beetle, development of new insecticides or breeding
resistant potato varieties alone will not suffice. It is
necessary that the population structure of the beetle
should receive due attention (as part of an integrated
system of protection of Solanaceae from this species).
The first step in this direction is to search and optimize
methods and approaches of finding and analyzing
polymorphism in this species, both in the phenotype
and in DNA (Udalov et al., 2003; Udalov et al., 2009).
The most frequently used tool for studying
intraspecific polymorphism in insect populations is
phenetic analysis. As discreetly changing traits, or
phenes, of the Colorado beetle, one can discern vari
ability in patterns of larval and imaginal coloration,
venation, and egg coloration. Of these various traits,
the phenes of imaginal coloration, specifically, phenes
of the vertex, occiput, pronotum, and elytra, are used
the most frequently (Kokhmanyuk, 1981; Fasulati,
Zeleev, 2002; Benkovskaya et al., 2004; 2008b).
In populations of such polymorphic species as the
Colorado potato beetle, there bound to be processes
leading to changes in the gene pool and, consequently,
phene pool. Examples can be found in works showing
changes in the phenetic structure of its populations
both over a long period of time (Eremina, Denisova,
1987; Ovchinnikova et al., 1984; Fasulati, 1985;
Zeleev, 2002) and in response to food supply: potato
varieties (Fasulati et al., 1994), other Solanaceae
plants (Fasulati, Karaseva, 1998; Benkovskaya, 2007),
and insecticides. The change in the phenetic structure
of the Colorado potato beetle populations in response
to the latter factor was shown for local populations of
Lipetsk oblast (Eremina, Denisova, 1987), Brest and
Lipetsk (Ovchinnikova et al., 1982; Klimets, 1988),
and Moscow and Lipetsk oblasts (Roslavtseva, Erem
ina, 2005). Unfortunately, in most of these works,
materials on polymorphism of only one phene were
used.
Thus, the following conclusions can be reached:
First, the analysis of the population structure of the
Colorado potato beetle geographic range (as well as
that of any other polymorphic and rapidly evolving
species) should be approached systematically if we are
to have a comprehensive idea of its temporal dynam
ics. Second, it is clear that the use of only one kind of
phenes—as in the abovementioned publications—is
not enough to successfully analyze the Colorado bee
tle population structure. Increasing the number of
phenes accounted for would provide a means of ana
lyzing the population structure by a greater number of
marker traits. Third, it is necessary to take into

390
 CHANGE IN THE POLYMORPHISM LEVEL
account different factors influencing the dynamics of
the species population structure in population studies.
Thus, the aim of our work was to assess the effect of
repeated exposure to insecticides as a factor influenc
ing the phenetic structure of the Colorado potato bee
tle population in the territory of the Republic of Bash
kortostan.
METHODS
Overwintered adults of the Colorado potato beetle
Leptinotarsa decemlineata Say were chosen as the
object of our research. They were collected by hand
from potato fields in a number of Bashkortostan
regions, 1–2 specimens per bush, diagonally, in the
intervals of 10–15 meters. This method produces a
more diverse sample than any other method (Amirkh
anov, 1995).
Phenetic polymorphism was analyzed using the
phenes of vertex (Klimets, 1997), occiput (Benk
ovskaya et al., 2004, 2008a, 2008b), pronotum (Fasu
lati, 1985), and elytra (Klimets, 1997). the susceptibil
ity of the Colorado beetle to insecticides was deter
mined using a spray solution of phoxim topically at a
dose of 1 μl/individual as described previously (Benk
ovskaya et al., 2008a).
The intrapopulation diversity was analyzed using
two indices suggested by L.A. Zhivotovskii—the aver
age number of variations μ and proportion of rare vari
ations h with their respective sampling errors sµ and sh:
2 others) should inevitably lead to a decrease in the
⎛ m
⎞ polymorphism (phenetic diversity) level in popula
μ = ⎜
⎝
∑ p m⎟ ,
⎠ tions.
1
μ(m – μ)
s µ ≈ ,
N
h = 1 – μ,
m less pronounced in elytra (Fig. 2). The average number
h(1 – h)
s h ≈ ,
N (1.9 times); for the pronotum phenes, from 8.38 ±
where m is the number of variations in the population,
p is the variation frequency, and N is the sample size
(Zhivotovskii, 1991).
The significance of differences between variation
frequencies was assessed using Student’s ttest (Lakin,
1990).
Calculation of all indices and visualization of the
obtained data were performed using the Microsoft
Excel 2003 software package. Population structures
were represented graphically as radar charts (Zhivot
ovskii, 1991) with radii lengths representing frequen
cies of different phene variations.
RESULTS AND DISCUSSION
The Colorado potato beetle first appeared in Bash
kortostan in 1976 in Kumertau and Arkhangel’skoe
RUSSIAN JOURNAL OF GENETICS: APPLIED RESEARCH
391
raions with the total area of nidi being 22 ha (Prognoz
poyavleniya i rasprostraneniya vreditelei …, 1978;
Migranov, 1994). Judging by the official reports, the
hotbeds were quickly eliminated, and the following sys
tematic inspections found no presence of the vermin. In
1977 a new hotbed, occupying 0.01 ha, was found in the
Al’sheevskii raion. By 1979 the Colorado beetle had
already colonized the whole territory of Bashkortostan.
We examined changes in the structure of local pop
ulations of the Colorado beetle on the territory of
Bashkortostan that have been going on for several
years. The Staromusino local population from Kar
maskaly raion was chosen as a research model; sam
ples were taken in 1994 and 2002.
Differences in the frequencies of phene variations
in samples from the given local population divided by
an eight year period are statistically significant for the
phenes of the vertex and pronotum patterns (p <
0.001), but uncertain for the elytra pattern.
As shown in Fig. 1, the changes observed between
1994 and 2002 are more distinct for the vertex and
pronotum than for the elytra. For example, the fre
quency of the m variation of the vertex phene increased
from 0.08 to 0.34, the ш, З, and M variations were not
found, the frequencies of 3, 6, and 9 variations of the
pronotum phene doubled, and the frequencies of 1
and 7 variations sharply decreased.
Such a tendency (decrease and disappearance of
some variations against the background of increase in
Indeed, the change in the phenetic diversity level of
the Colorado potato beetle was registered in the exam
ined local population. Such changes were more pro
nounced (differences were significant when p < 0.001)
in the phenes of the vertex and pronotum patterns and
of variations μ for the vertex phene decreased from
3.64 ± 0.157 in 1994 to 1.95 ± 0.291 in 2002
0.162 to 6.41 ± 0.49 (1.3 times); and for the elytra
phenes, from 2.30 ± 0.174 to 1.92 ± 0.291 (1.2 times).
The opposite tendency is noted for the other index,
the proportion of rare variations h that characterizes
the structure of intrapopulation diversity (Fig. 2).
Between 1994 and 2002, this index value for the vertex
phene increased 2.2 times (from 0.27 to 0.61); for the
pronotum, it was 4.2 times (from 0.07 to 0.29); and for
elytra the proportion of rare variations remained more
or less the same, with a tendency to increase. In our
opinion, such changes are explicable on the basis of
the fact that in the process of formation of this local
population (similar tendencies are typical for samples
from other local populations), the decrease in diversity
was caused by redistribution of variation frequencies,
in the course of which some variations, formerly com
prising the “skeleton” of the population phenetic
Vol. 1 No. 5 2011
392 UDALOV, BENKOVSKAYA

A B C
O 1
0.8 W
9 2 1.0
0.2
0.4 0.1 0.5
1994 M m
8 3
0 0 0

7 4 W/V V
З ш
6 5
1
O W
9 2 1.0
0.8 0.2
0.4 0.1 0.5
M m 8 3
2002 0
0 0
7 4 W/V V
З ш
6 5

Fig. 1. Changes in the phenetic structure of the Staromusino local population of the Colorado potato beetle over eight years.
A, variations of the vertex phene; B, pronotum phenes; C, elytra phenes. Frequencies of phene variations are marked against the
respective axes.

A B C
5 5
Average number
of variations, µ

4 9 4
3 3
6
2 2
3 1
1
0 0 0
1994 2002 1994 2002 1994 2002
rare variations, h

0.7
Proportion of

0.6 0.4 0.7

0.5 0.6
0.4 0.3 0.5
0.3 0.4
0.2 0.3
0.2 0.2
0.1 0.1 0.1
0 0 0
1994 2002 1994 2002 1994 2002

Fig. 2. Change of the level of phenetic diversity in the Staromusino local population of the Colorado potato beetle over eight years. A,
variations of the vertex phene; B, pronotum phenes; C, elytra phenes. Data are given with their respective sampling errors sµ and sh.

appearance, became rare. The variations that have
been rare from the outset disappear as a result of the
selection washing them away in the course of insecti
cide treatment.
Another hypothesis is that the most significant fac
tor influencing the phenetic structure of the Colorado
potato beetle population on the territory of the South
Urals was the selective effect of insecticides. In the
course of this unconscious selection (in the terms of
Yablokov, Yusufov, 2004), selective survival of individ
uals resistant to the effects of insecticide and elimina
tion of susceptible individuals, whose genotypes are

RUSSIAN JOURNAL OF GENETICS: APPLIED RESEARCH Vol. 1 No. 5 2011

 CHANGE IN THE POLYMORPHISM LEVEL 393

Average number of variations, µ Proportion of rare variations, h

A
5 1.0
4 0.8
3 0.6
2 0.4
1 0.2
0 0
C T C T
B
10 1.0
8 0.8
6 0.6
4 0.4
2 0.2
0 0
C T C T
C
5 1.0

4 0.8

3 0.6

2 0.4

1 0.2
0
0 C
C T T

Fig. 3. Change of the level of phenetic diversity in the Kazangul local population of the Colorado potato beetle according to the
results of laboratory experiments. A, variations of the vertex phene; B, pronotum phenes; C, elytra phenes. White bars, survivors;
grey bars, eliminated individuals. C, control; E, experiment. Data are given with their respective sampling errors sµ and sh.

marked by certain pattern phenes, takes place. We
noted previously (Udalov, 2006; Benkovskaya et al.,
2006) that the level of the resistance to insecticides and
frequencies of some phene variations in local popula
tions of the beetle in the Republic of Bashkortostan
rise conjointly. Specifically, individuals with pronotum
variations 3, 6, and 9 prevailed in samples after insec
ticide treatment (Benkovskaya et al., 2004, 2008a).
Most likely, the doubled share of these variations in the
sample is explained by selective survival of individuals
possessing these phenotypes due to their increased
resistance to insecticides (Fig 1B). A similar pro
cess—increase of the proportion of 3, 6, and 9 phene
variations of the pronotum pattern—was observed in
populations of the Colorado beetle in other regions:
Moscow oblast (Roslavtseva, Eremina, 2005), Bry
ansk (Oleinikov et al., 2006), and Kaliningrad, Rostov,
and Vologda oblast (Vasil’eva et al., 2005).
We also examined the influence of insecticides on
change in the diversity level in samples from the Kaza
ngul local population (Davlekanovo raion, Bashkor
tostan) of the Colorado potato beetle in the laboratory
environment.
Figure 3 shows changes in the phenetic diversity
levels of the beetle in total samples and samples that
survived treatment with a lethal concentration of vola
ton in laboratory experiments.
Statistically significant were only changes in fre
quencies of the pronotum phene variations when
comparing survivors in the experiment with survivors
in the control and survivors in the experiment with
those eliminated in the experiment (p < 0.001). The
comparison of the samples with respect to the fre
quencies of vertex and elytra phenes did not produce
statistically significant results.

RUSSIAN JOURNAL OF GENETICS: APPLIED RESEARCH Vol. 1 No. 5 2011

394 UDALOV, BENKOVSKAYA
A decrease in the phenetic polymorphism level (the
average number of variations μ) in the group of survi
vors as compared both to the control (1.7 times for the
vertex phenes, 1.5 times for the pronotum, and 2.3 for
the elytra phenes) and to the beetles eliminated by the
treatment (1.9 times for the vertex phenes, 1.4 times
for the pronotum, and 2.5 for the elytra phenes) was
observed.
CONCLUSION
Thus, we analyzed the differences in phenetics of a
number of local populations of the Colorado potato
beetle on the territory of the republic of Bashkortostan
gathered in 1994 and 2002. Certain changes in the phe
netic structure of local populations of the beetle were
found. A decrease in the phenetic polymorphism level
(average number of variations) as a result of repeated
treatment with insecticides was observed in nature and
verified by laboratory experiments. This lets us believe
that the changes in the polymorphism level of the Col
orado potato beetle populations in the South Urals are
caused mainly by this anthropogenic factor.
ACKNOWLEDGMENTS
This work was supported by the Russian Founda
tion for Basic Research, project nos. 090400391a
and 110401886a and 110497022r_ povolzh’e _a.
If fellow entomologists could send us specimens of the
Colorado potato beetle from other parts of its geo
graphic range, we would greatly appreciate it. We are
grateful to the reviewer of our manuscript for the eval
uation and for valuable constructive suggestions.
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