10 - CMB-2014v4n11页

Computational Molecular Biology 2014, Vol. 4, No. 12, 1-8
http://cmb.biopublisher.ca
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Optimum temperature of amplification was found to
be 570C and 580C as amplified band found to be
deeper as compared to other annealing temperature.
This indicated that the primer pair Dep1-1 (Promoter)
for Dep1 gene would be useful for introgressing genes
from high grain varieties into low grain varieties in
order to improve yield potential through molecular
breeding approach.
The primer pair Dep1-2 (Exon1) for Dep1 gene
amplified band of 600bp in all the six temperatures
from both high and grain varieties, HGN1 and Heera,
respectively, though variation was found in strength of
amplified bands. Optimum temperature of
amplification was found to be 570C and 580C as
amplified band found to be deeper as compared to
other annealing temperature (Figure 8).
Figure 8 Gradient PCR with Dep1 gene specific primer,
Dep1-2 (Exon1) 1 - HGN1 (high grain), 2- Heera( low grain)
Note: Annealing Temperatures are given on the top of gel
This indicated that the primer pair Dep1-2 (Promoter)
for Dep1 gene would be not be useful for
introgressing genes from high grain varieties into low
grain rice varieties as there is no difference in length
polymorphism. However, further investigation is
necessary to find out whether there is difference in
restriction site resulting in fragment size variation.
The primer pair Dep1-3 (Exon2) for Dep1 gene
amplified band of 600bp in all the six temperatures
from both high and grain varieties, HGN1 and Heera,
respectively, though variation was found in strength of
amplified bands in HGN1 and Heera. Optimum
temperature of amplification was found to be 560C,
570C and 580C as amplified band found to be deeper
as compared to other annealing temperature (Figure
9).
This indicated that the primer pair Dep1-3 (Promoter)
for Dep1 gene would be not be useful for
introgressing genes from high grain varieties into low
Figure 9 Gradient PCR with Dep1 gene specific primer, Dep1-3
(Exon2) 1-HGN1 (high grain), 2- Heera( low grain) Annealing
Temperatures are given on the top of gel
grain rice varieties as there is no difference in length
polymorphism. However, further investigation is
necessary to find out whether there is difference in
restriction site resulting in fragment size variation.
The primer pair Ghd-7 (Exon1) for Ghd1 gene
amplified band of 650bp in all the six temperatures
from high grain variety, HGN1 while fail to amplify
from Heera (Figure 10).
Figure 10 Gradient PCR with Ghd7 gene specific primer,
Ghd7-2 (Exon1) 1 - HGN1 (high grain), 2- Heera( low grain)
Annealing Temperatures are given on the top of gel
All the six temperature of amplification were found to
be optimum as amplified band found to be medium to
deeper at all six annealing temperature. This indicated
that the primer pair Ghd-7 (Exon1) for Ghd1 gene
would be not be useful for introgressing genes from
high grain varieties into low grain rice varieties as
there is no difference in length polymorphism.
The primer pair Apo1-1 (promoter) for Apo1 gene did
not amplify any band at the six temperatures from
either high or low grain varieties, HGN1 and Heera,
respectively indicating that the primer pair Apo1-1
(promoter) for Apo1 gene would be not be useful for
introgressing genes from high grain varieties into low
grain rice varieties (Figure 11) (Table 2 and 3).