ISSN 1022-7954, Russian Journal of Genetics, 2008, Vol. 44, No. 3, pp. 320–325. © Pleiades Publishing, Inc., 2008.
Original Russian Text © E.A. Levina, I.Yu. Adrianova, G.D. Reunova, Yu.N. Zhuravlev, 2008, published in Genetika, 2008, Vol. 44, No. 3, pp. 374–380.
According to various literature sources, several
A. Henry [1–3] occur
on the territory of Primorye. According to a number of
authors, of all these species, only Olga Bay larch
does not have a hybrid origin, whereas the
remaining forms may be assigned to a complex that
resulted from introgressive hybridization [4, 5].
Hybridization leads to high variability of morphologi-
cal traits and to controversial views in taxonomy of
larches from Primorye.
is among the most ancient larches.
This species is a Pliocene relict  listed in the Red
Book of the Russian Federation . In the Russian Far
East, it occupies a small area in the southeastern Pri-
morye stretching along the coast of the Sea of Japan
and eastern slopes of Sikhote-Alin’ . As the compet-
itive ability of
is lower than that of other for-
est species, including hybrid larches, the former occurs
in small isolated patches , which creates the risk of
is a putative parent of a number of
hybrid complexes of larches from the south of the Rus-
sian Far East, which arose in Pleistocene–Holocene ,
it seems important to study genetic variation and the
population structure of this species using molecular
DNA markers, which permit, in contrast to allozyme
analysis, to examine variation of both coding and non-
coding DNA regions. One of such methods used in pop-
ulation genetics of conifers is DNA ampliﬁcation by
primers with random nucleotide sequences (random
ampliﬁed polymorphic DNA, RAPD) [9–15]. Using
the RAPD method, genetic variability of larches from
various geographic regions of Siberia and the Russian
Far East and their genetic relationships were evaluated
The aim of this study was studying genetic variabil-
ity and establishing genetic relationships among the
natural larch populations within the range of
in Primorye based on RAPD marker analysis.
MATERIALS AND METHODS
In our study, we used needles of nine to twelve trees
from each of the ten samples examined (Fig. 1). The
samples are described in Table 1. Genomic DNA was
isolated from 300–500 mg of fresh needles following
the protocol from  with slight modiﬁcations.
Polymerase chain reaction was run in a thermal
cycler UNO II 48 (Biometra, Germany) with decamer
random primers (Operon Technologies, United States;
SibEnzyme, Novosibirsk), using the reaction mixture
and temperature conditions described in . The con-
trol probe contained the total ampliﬁcation mixture
without DNA. Each reaction was run in two to four rep-
licates. The ampliﬁcation products were fractionated by
electrophoresis in 1.4% agarose gel in the presence of
ethidium bromide and photographed in UV light. To
assess size of the fragments (amplicons),
restriction fragments of phage lambda DNA (Fermen-
tas, Lithuania) were used as molecular mass markers.
Genetic Variability and Differentiation in the Larch Populations
within the Range of
A. Henry in Primorye
E. A. Levina, I. Yu. Adrianova, G. D. Reunova, and Yu. N. Zhuravlev
Institute of Biology and Soil Sciences, Russian Academy of Sciences, Vladivostok, 690022 Russia;
Received December 13, 2006
—The level of within- and among-population variation of larch inhabiting the range of
A. Henry in Primorye was estimated on the basis of 440 RAPD loci identiﬁed by means of 12 random primers.
In ten populations examined, the proportion of polymorphic loci was 35–60%, the average expected heterozy-
gosity varied from 0.1340 to 0.2169, and the average gene ﬂow estimate was 1.38. According to Fisher’s test
for heterogeneity, the pairwise differences of the fragment frequencies between the populations were statisti-
cally signiﬁcant. The subdivision index
= 0.2663 indicated that the interpopulation variation component
accounted for approximately 27% of the total variation. Coefﬁcients of Nei’s genetic distance between the pop-
ulations varied from 0.0137 to 0.0934. Most of the samples with high support clustered according to the geo-
graphic positions relative to one another within the range. These results suggest that the populations examined
are characterized by high genetic variation, like the larch populations of Siberia and the Russian Far East stud-
ied earlier, but, in contrast to the latter, exhibit higher among-population variability.