Triticeae Genomics and Genetics 2015, Vol.6, No.2, 1-7
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Table 1 Pedigree and origin details of bread wheat genotypes
S No. Genotypes Salinity tolerance Pedigree
Origin
1.
MP-4010
AMGOSTURA 88 (CM 50123-3M-Y-3M-1Y-2M-Y-
2M-2Y-0M-OMR/S)
JNKVV, Jabalpur, Madhya Pradesh
2.
HI- 8713
HD 4672/PBW 233
Indore, IARI, REG. RES. Station
3.
C-306
RGN/CSK 3//2* C 591/3/C 217/N 14//C 281
CCSHAU, Haryana
4.
Lok-1
S-308/S-331
Gujrat
5.
Raj-4037
DL 788-2/Raj 3717
RAU, Durgapura
6.
HD-2932
KAUZ/STAR/HD 2643
IARI, New Delhi
7.
KH-65
Selected from KHARCHIA local
RAU, Durgapura
8.
KRL-19
PBW 255/ KRL 1-4
CSSRI, Karnal
9.
KRL-213
CNDO/R143//ENTE/MEX1-2/3/AEGILOPS
SQ4ARROSA (TA4S)/4/WEAVER/5/2/* KAUZ
CSSRI, Karnal
10.
KRL-210
PBW65/2*PASTOR
CSSRI, Karnal
11.
KRL-1-4
KHARCHAI 65/WL 711
CSSRI, Karnal
12.
Raj-3765
HD 2402/VL 639
RAU, Durgapura
out for 24 hr with four changes of water at interval of
6 hr. A small quantity of precipitate formed at this
stage was removed by centrifugation and supernatant
were freezed.
The freeze supernatants lyophilized in separate petri
plates and dried samples immediately packed in
airtight bottles. The known amount of protein
dissolved in phosphate buffer (pH 7.4) and subjected
to vertical gel electrophoresis (SDS-PAGE).
The variability of seed storage-proteins was analyzed
by using SDS-PAGE (Damania et al., 1983).
1.3 Preparation of separating gel (15% and 10%)
and stacking gels
The 15% gel was prepared by mixing 20 ml of stock
acrylamide solution, 8.0 ml tris HCl (8.9) and 11.4 ml
of water. Subsequently 0.2 ml of ammonium per
sulphate solution, 0.4 ml 10% SDS and 20 µl
Tetramethylethylenediamine (TEMED) was added.
The 10% gel was prepared by mixing of 13.3 ml of
stock acrylamide solution, 8 ml tris HCl (8.9) and 18.1
ml of water. Subsequently 0.2 ml of ammonium per
sulphate solution, 0.4 ml 10% SDS and 20 µl TEMED
was added. The 3% stacking gel was prepared by
addition of 4.0 ml of stock acrylamide solution, 2.0 ml
tris HCl (6.7), and 2.0 ml of riboflavin solution and
8.0 ml of water. And this solution was mixed with
0.1 ml 10% SDS and 20 µl TEMED.
1.4 Preparation of sample and casting of gel
Protein sample (48 µl ) was mixed with 12 µl of 4x
sample buffer (0.25M Tris-HCl pH 6.8 added with 8%
SDS, 40% glycerol, 20%
-mercaptoethanol, 0.5%
bromophenol blue) and heated in water bath
containing boiling water for 2-3 min to ensure
complete interaction between protein and SDS.
Sample was cooled to room temperature.
1.5 Data analysis
The photographs of SDS-PAGE gel was used to study
the protein profile of the all genotypes. The bands
were designated on the basis of their molecular weight.
The presence of protein band was scored as “1” and
its absence as “0” (Table1, S-1 and S-2). Only bright,
clearly distinguishable bands were used in genetic
analysis. In the present study, the population diversity
based on SDS PAGE banding patterns was calculated
using POPGENE 1.31 (Yeh et al., 1999) software. The
data matrices were then entered into NTSYS-PC
(Numerical Taxonomy and Multivariate Analysis
System Program) developed by Rholf (2000). The
data were analyzed using SIMUQUAL Jaccard’s
Similarity Coefficients formula (Sneath and Sokal,
1973). The matrix of similarity coefficients generated
from data of 12 wheat genotypes was subjected to un
weighted pair group method for arithmetic average
(UPGMA) using NTSYS-PC, version 2.02 (Exeter
software, New York).
2 Results and Discussion
The variability of seed storage-proteins was analyzed
by using SDS-PAGE (Damania et al., 1983). The
study comprises characterization of high and low
molecular weight glutenin subunits of different wheat
varieties by SDS-PAGE and assessment of genetic
