ISSN 1022-7954, Russian Journal of Genetics, 2009, Vol. 45, No. 7, pp. 805–810. © Pleiades Publishing, Inc., 2009.
Characterization of genetic diversity in crop species
has long been based on morphological traits, however,
morphological variation is often found to be of limited
because expression of morphological traits may be
affected by environmental conditions, thereby con-
straining the analysis of genetic variation. During
resent year, biochemical and molecular genetic tech-
niques have emerged as a complementary strategy in
conjunction with traditional approaches in the manage-
ment of plant genetic resources [1, 2].
For studies on population genetic structure; protein
assays are considered as useful tool for most taxa [2–5].
Sodium Dodecyl Sulphate Polyacrylamide Gel Electro-
phoresis (SDS-PAGE) is widely used due to its reliabil-
ity and simplicity in describing the genetic structure of
crop germplasm . A considerable variation in protein
banding pattern has been reported, which was localized
to various geographical regions .
The present study is the ﬁrst report on Pakistani
pea’s germplasm seed storage for investigation of
diversity in relation to geographical pattern.
MATERIALS AND METHODS
Seeds of ninety seven genotypes were collected
from all over Pakistan (
For SDS-PAGE analysis, single seed from each geno-
type was ground to a ﬁne powder with mortar and pes-
The article is published in the original.
tle. Four hundred
l protein extraction buffers (0.5 M
HCl (pH 6.8), 2.5% SDS, 10% glycerol and 5%
2-mercaptoethanol, was added to 0.01 g of seed ﬂour
and mixed well with Automatic Lab-Mixer (DH-10).
Bromophenol blue (BPB) was added to the protein
extraction buffer as tracking dye to monitor the move-
ment of protein in the gel. The SDS-PAGE of total seed
protein was carried out in the discontinuous buffer sys-
tem as describe by . It was observed that 15% acry-
lamide gel concentration with
l of sample gave the
best resolution. After staining and destaining, the gel
was dried using gel drier (Atto, Rapidery-Mini Japan).
In order to check the reproducibility of the method two
separate gels for all samples were run under similar
electrophoretic condition. The molecular weight of the
dissociation polypeptides was determined using molec-
ular weight protein sanders”MW-SDS-70 Kit (Sigma,
United States). For scoring, electrophoregram was
divided into three regions and data was scored for the
presence or absence of protein bands.
The SDS-PAGE data were analysed for cluster using
computer packages STATISTICA 6.0 in Windows XP-
2005. The data were analysed on the basis of Province
Wise (PWA) and Agro-Ecological Zones (AEZ) analy-
sis . A similarity index was used to construct a den-
drogram by the UPGMA method .
45 – >66.0
First Proteomic Assay of Pakistani
Germplasm Relation to Geographic Pattern
, A. Ghafoor
, M. R. Khan
, and Asmatullah
Department of Botany, University of Malakand, NWFP, Pakistan
Plant Genetic Resources Program (PGRP), National Agricultural Research Centre (NARC), Islamabad, Pakistan
Department of Biochemistry, Quaidi-Azam University Islamabad, Pakistan
Department of Botany, University of Science and Technology Bannu, NWFP, Pakistan
Received March 11, 2008
—Proteomic assay was carried out to asses genetic diversity in relation to geographic pattern in
97 genotypes of
L., collected from all over Pakistan. In total 34 bands were observed and among
these, 26.7% bands were monomorphic, while 73.5% bands showed polymorphism. Based on both Province
Wise Analysis (PWA) and Agro-ecological Zones (AEZ) the genotypes collected from Punjab, North West
Frontier Province (NWFP) exhibited 70.6%, and 64.7% variation respectively. The germplasm collected from
Azad Kashmir showed the lowest level of genetic diversity. Cluster analysis exhibited, moderate level of asso-
ciation, between genetic diversity and geographic pattern of the genotypes.