ISSN 1022-7954, Russian Journal of Genetics, 2009, Vol. 45, No. 7, pp. 788–793. © Pleiades Publishing, Inc., 2009.
Original Russian Text © O.P. Dribnokhodova, S.A. Gostimsky, 2009, published in Genetika, 2009, Vol. 45, No. 7, pp. 900–906.
It is topical to develop new methods for studying
molecular genetic polymorphism of cultivated plants .
Simple sequence repeat (SSR) analysis allows a
simple and rapid genotyping of the accessions of inter-
est at microsatellite loci. Microsatellites are simple
repetitive sequences with a unit of 2–6 bp that form
more of less regular tracts of up to several hundreds of
base pairs. Microsatellites are widespread in the
genomes of all eukaryotes and are uniformly distrib-
uted through all chromosomes . Thus, SSRs provide
one of the most polymorphic and highly informative
marker systems [3, 4]. SSRs are currently used in basic
and applied studies to construct genetics maps; to local-
ize genes; to compare higher plant species; to examine
plant, animal, and human populations [5, 6]; to estimate
genetic diversity; and to identify the genotype .
Although many microsatellite loci have been identiﬁed
genome , its interlinear poly-
morphism is poorly understood.
The objective of this work was to study the genetic
varieties, lines, and mutants with
the use of microsatellite markers.
MATERIALS AND METHODS
We examined 40
20 varieties, 12 marker lines, and 8 mutants from the
collection of the Department of Genetics, Moscow
State University. Total DNA was isolated from young
leaves by the CTAB–isopropanol method  with mod-
The analysis was performed with 22 primer pairs,
which allowed ampliﬁcation of 23
ellite loci. Primer AA255 allowed ampliﬁcation of two
nonlinked microsatellite loci. The primer sequences
were as in ; the reaction conditions were selected
experimentally. The primer sequences, annealing tem-
peratures, and recommended electrophoretic condi-
tions are summarized in Table 1.
Ampliﬁcation was carried out in a Tertsik thermal
cycler and included initial denaturation at
2 min 30 s; ﬁve cycles of
for 30 s,
for 30 s, and
for 1 min 20 s; 35 cycles of 93
for 20 s,
C for 30 s, and 71.5
C for 1 min; and last
synthesis at 72
C for 5 min. The annealing temperature
was varied from 55 to 67
C depending on the primer
(Table 1). The ampliﬁcation products were electro-
phoretically resolved in 3% agarose gel in the case of
fragments of more than 250 bp and in 6% polyacryla-
mide gel in the case of fragments of less than 250 bp,
with subsequent ethidium bromide staining.
RESULTS AND DISCUSSION
For the ﬁrst time in Russia, the allele composition of
23 microsatellite loci was studied in 40
accessions. Each of the accessions had a unique allele
combination, and the alleles were well distinguishable
in repetitive examinations.
All of the loci under study proved to be highly poly-
morphic; the number of alleles per locus varied from 2
to 10, averaging 4.65. The polymorphism information
= 1 –
is the frequency
-th allele of the
-th locus) varied from 0.139 to
0.816, averaging 0.620 (Table 2). This ﬁnding agreed
well with published data for pea (0.63 ) and other
higher plant species (0.53 for maize ). The linkage
group, allele number, and polymorphism information
content for each of the loci are summarized in Table 2.
The approximate size of the shortest ampliﬁed allele is
Figure 1 shows the patterns obtained with primer
AD160 for some of the lines, varieties, and mutants. In
Allele Polymorphism of Microsatellite Loci in Pea
L. Lines, Varieties, and Mutants
O. P. Dribnokhodova and S. A. Gostimsky
Department of Genetics, Moscow State University, Moscow, Russia;
Received November 28, 2007; in ﬁnal form, July 23, 2008
—Interlinear polymorphism at 23 microsatellite loci was studied in 40
and mutants and proved to be high, 61.6% on average. Varieties bred for different end uses substantially differed
in the extent of polymorphism and allele composition. Polymorphism of microsatellite loci was shown to be
suitable for developing passports of industrial pea varieties.