1022-7954/04/4007- © 2004
Russian Journal of Genetics, Vol. 40, No. 7, 2004, pp. 737–742. Translated from Genetika, Vol. 40, No. 7, 2004, pp. 909–915.
Original Russian Text Copyright © 2004 by Cheghamirza, Koveza, Konovalov, Gostimsky.
The garden pea (
L.) is an important
agricultural plant with a rich history of genetic research.
The wide morphological variation which
= 14) exhibits has provided genetic markers for pio-
neering geneticists and formed the basis for earlier link-
age maps . Linkage maps of the pea genome contain-
ing molecular markers have also been published [2, 3].
Through the use of anchor loci, the map has been
related to the “standard map” of the pea genome ,
which contains both molecular and morphological
markers. However, the genetic saturated map of pea
was not prepared yet and thus complementary search is
needed to identify new molecular and morphological
The RAPD (random ampliﬁed polymorphic DNA)
technique is simple, quick, relatively inexpensive, and
suitable to process many samples per day, making it
potentially interesting for genetic mapping . Never-
theless, poor sensitivity and a lack of reproducibility
are also associated with this technique. In order to solve
these problems, the informative RAPD fragments can
be converted into SCAR markers (sequence character-
ized ampliﬁed regions) [5, 6], which generate a simple,
reliable, and sometimes codominant, banding pattern.
The SCAR markers can serve as anchor points between
physical and genetic maps.
In the ISSR (intersimple sequence repeat) tech-
nique, single simple sequence repeat (SSR) primers,
which are composed of short nucleotide repeats such as
those found in microsatellites, are used in the poly-
merase chain reaction (PCR) .
CAPS (cleaved ampliﬁed polymorphic sequence)
markers have a number of advantages over RAPDs and
other molecular markers. The CAPS markers are
codominant, unlike the RAPDs and ISSRs. CAPS poly-
morphisms are detected by a PCR assay followed by
restriction–endonuclease digestion; therefore, they use
little template DNA and are rapid and relative inexpen-
sive to assay . CAPS markers were employed to map
pea genome .
In this paper, we report RAPD, ISSR, and CAPS
markers linked to a new chlorophyll mutation (
gene) and the location of this new mutation.
MATERIALS AND METHODS
population of 223 individuals was developed
by crossing two
genotypes, WL1238 and
Chi115. The female parent, Chi115, is a chlorophyll
mutant induced by ethylmethane sulfonate (EMS)
treatment of seeds of genotype Torsdag; it is character-
ized by lighter plant color. The male parent, WL1238,
is a tester line, often exploited in genetic studies.
plants of each F
plant showing the
wild-type phenotype were analyzed to distinguish their
heterozygous or homozygous dominant genotypes.
To establish a local map around the
RAPD and ISSR techniques were used with 45 RAPD
Identification and Mapping of
Gene and DNA Markers
Linked to It in Pea (
, O. V. Koveza
, F. A. Konovalov
, and S. A. Gostimsky
Agronomy and Plant Breeding Department, Razi University, Kermanshah, Iran;
fax: (095) 939-35-12; e-mail: email@example.com
Department of Genetics, Moscow State University, Moscow, 119899 Russia
Received January 8, 2004
—Chlorophyll mutant Chi115 was induced by ethylmethane sulfonate (EMS) treatment of seeds of
genotype Torsdag in Moscow State University and is characterized by lighter plant color. The monogenic nature
of the mutant was determined by analyzing the F
population from a cross between two
WL1238 and Chi115. To establish a local map around the
gene, the RAPD and ISSR techniques were
used with 45 RAPD and 10 ISSR primers in combination with bulked segregant analysis (BSA). Linkage of 12
RAPDs and 2 ISSRs to the
locus was observed in analysis of F
single plants. Two RAPD markers that
were closely associated with the
gene were converted into the sequence characterized ampliﬁed region
(SCAR) markers. By lowering the LOD score to 2, the linkage group containing the
gene could be linked
gene (color of the ﬂower) on linkage group III. Nevertheless, to prove the result obtained, three CAPS
markers Sodmt, TubA1, and Rb were chosen on linkage group III. The results of linkage analysis showed that
these CAPS markers were located within the linkage group including the