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International Journal of Horticulture 2014, Vol.4, No.10, 50
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52
http://ijh.biopublisher.ca
51
Figure 1 PCR amplification of DNA from Resistance and
Susceptible parents using SCAR marker SK 14
Note: AA: Arka Anoop
Screening of F
1
population for rust resistance
F
1
’s obtained from each cross combination were raised
in field and at first trifoliate leaf stage, DNA was
extracted. F
1
’s were screened using molecular marker,
SK 14. F
1
’s involved IC525236 as resistant parents
showed the resistant banding pattern 620 bp (Table 1;
Figure 1).
Screening of F
2
population for rust resistance
SCAR marker, SK 14 assisted foreground selection
for rust resistant gene was done in the F
2
(Table 2;
Figure 2). Initially, the parental DNA with SCAR
markers linked to rust resistance was amplified. The
PCR conditions for SCAR primers were optimized for
DNA amplification to obtain a good amplification
with clear bands at optimum annealing temperature.
SK 14
620
amplified at 620 bp in resistant parent
IC-525236 and no amplification was observed in
IIHRPB-7 (susceptible). In F
2
population from the
cross IIHRPB-7 X IC-525236, out of 100 plants
screened, 71 plants showed the resistant banding
pattern at 620 bp indicating the presence of target
gene. These resistant lines were selected for further
advancing. This resistant marker was also used for
screening F
1
population, wherein all the lines showed
the presence of the above marker associated with rust
resistance. Similarly, Vikas
et al.
(2012) incorporated
blast resistance genes in rice
viz.,
Piz-5 and Pi54, from
the donor lines C101A51 and Tetep into the genetic
background of PRR78 through marker assisted
foreground selection. Foreground selection for these
genes was done using tightly linked molecular
markers, AP5930 and RM206. But even though
IIHRPB-1 and IIHRPB-2 were highly susceptible to
rust showed amplification as like in resistant parent
IC-525236 at 620 bp. This showed that resistance to
individual rust races in bean line IC-525236 (
Ur-3)
is
conditioned by single dominant gene linked in
coupling that appear to be inherited as a complex
Figure 2 PCR amplification of DNA from F
2
individuals of
IIHRPB-7×IC-525236 using SCAR marker SK 14
Table 2
χ
2
estimates for rust resistance in several crosses of F
2
population
Observed frequency
Expected frequency
No. F
2
genotypes
Resistant
Susceptible
Resistant
Susceptible
Total
2
value P value
1 IIHRPB-7 X IC-525236 71
29
75
25
100 0.21
0.71
2
(p=0.05)
3.841