1022-7954/02/3807- $27.00 © 2002
Russian Journal of Genetics, Vol. 38, No. 7, 2002, pp. 785–790. Translated from Genetika, Vol. 38, No. 7, 2002, pp. 937–943.
Original Russian Text Copyright © 2002 by Popov, Urbanovich, Kirichenko.
Classiﬁcation and identiﬁcation of genotypes in
sunﬂower is an important task in sunﬂower breeding. In
most cases, the identiﬁcation of sunﬂower cultivars,
lines and hybrids is based on morphological traits.
However, these traits do not always enable to distin-
guish between closely related accessions, to detect hid-
den genetic variability, and to control homogeneity of
the inbred lines. Moreover, morphological traits may be
unstable, and their number is limited . Using molec-
ular-genetic and protein markers for these purposes
proved to be more effective [1, 2]. Molecular-genetic
markers have many advantages over morphological
traits including higher information content, which is of
primary importance for systematization and registra-
tion of collection specimens.
Molecular-genetic markers are widely used in such
cultures as wheat [2–4], barley [5, 6], buckwheat ,
and many others [2, 8] including sunﬂower. The sun-
ﬂower genome has been currently examined by means
of restriction fragment length polymorphism analysis
(RFLP markers), polymerase chain reaction with ran-
dom primers (RAPD markers), and ampliﬁed fragment
length polymorphism analysis (AFLP markers). Using
molecular-genetic markers one can examine genetic
diversity of species, cultivars, lines and forms of sun-
ﬂower, identify genotypes, systematize collection
material [9–13], classify heterosis groups , and
construct genetic maps . For instance, studies of
inter- and intraspeciﬁc differentiation of the genus
and the establishment of genetic distances
between selection inbred lines of sunﬂower were based
on RAPD analysis [16, 17].
This work was aimed at studying genetic diversity
of classiﬁcation of inbred lines of sunﬂower using
RAPD and isozyme analyses.
MATERIALS AND METHODS
As plant material, we used 30 inbred sunﬂower lines
selected at the Yurjev Plant Production Institute. Four-
teen of these lines (Kh908, Kh503, Kh1002, Kh1006,
Kh1007, Kh1008, Kh3848, Kh4353, Kh1010, Kh2111,
Kh2552, Kh2122, Kh4021, and Kh1012) were used as
They were created by selection from interspeciﬁc
hybrids and cultivars and selected for high productivity.
Lines Kh984, Kh983, Kh785, Kh787, Kh991, Kh711,
Kh818, Kh840, Kh943, Kh821, KhX767, Kh954,
Kh726, Kh945, Kh769, and Kh956 are used in sun-
ﬂower breeding as paternal forms (fertility restorers).
These lines mostly originated from American and
Yugoslavian cultivars and were selected for disease
DNA was isolated from six to ten ripe seeds as
described in . The DNA amount was determined in
agarose gel using a calibration curve constructed on the
basis of assaying specimens with known amount of
DNA. The DNA content was estimated using the Quan-
tity One program.
RAPD analysis was conducted with 12 random
primers listed in the table. Primers P36, P37, P46, P53
were chosen for detecting polymorphic loci in sun-
ﬂower . Primers OPT-08, OPW-04, OPW-06,
OPW-09, OPW-10, OPW-15, and OPX-01 were pur-
chased from Oregon Technologies (United States).
The polymerase chain reaction (PCR) was conducted
in the OmniGene thermal cycler (Hybaid, England).
A reaction mixture (30
l) contained 50 mM KCl;
10 mM Tris–HCl, pH 9.0; 0.1% triton X-100; 2.5 mM
; 0.2 mM of each of dATP, dCTP, dGTP, dTTP;
m of the primer; 30 ng of DNA; and 1.5 units of
polymerase. The reaction was conducted as fol-
lows: ﬁrst denaturation for 5 min at 94
C; 45 cycles of
1 min at 94
C, 1 min at 36
C, and 2 min at 72
Studying Genetic Diversity in Inbred Sunflower Lines
by RAPD and Isozyme Analyses
V. N. Popov
, O. Yu. Urbanovich
, and V. V. Kirichenko
Yurjev Plant Production Institute, Ukrainian Academy of Agrarian Sciences, Kharkov, 61060 Ukraine;
fax: (0572)92-03-54; e-mail: firstname.lastname@example.org
Institute of Genetics and Cytology, National Academy of Sciences of Belarus, Minsk, 220072 Belarus
Received July 31, 2001; in ﬁnal form, December 12, 2001
—Genetic diversity of 30 inbred sunﬂower lines was examined by RAPD and isozyme analyses. The
inbred lines were shown to be highly polymorphic by RAPD markers. The line distribution on genetic similarity
dendrograms based on the RAPD and isozyme data was analyzed. High effectiveness of RAPD analysis for dif-
ferentiating genotypes of inbred sunﬂower lines was demonstrated.