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Molecular Plant Breeding Provisional Publishing
Molecular Plant Breeding 2012, Vol.3, No.5, 50
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56
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52
Figure 2 Calli response on regeneration selection medium (RSM) at various levels of basta
Note: A: 1 mg/L; B: 3 mg/L; C: 5mg/L; D: 7mg/L; E: 10 mg/L
1.2 Selection of optimal basta dose for genotype
S-2003-us-127 to be used in regeneration selection
media for transformation experiment
Calli of genotype S-2003-us-127 were induced on
CIM2 (Table 1) and 28 days old healthy and
proliferated calli were transferred on regeneration
selection media (RSM) in which five levels of basta
(1 mg/L, 3 mg/L, 5 mg/L, 7 mg/L, 10 mg/L) were
used (Table 2) followed by incubation at (26±1)
℃
under 16/8 hrs light dark period. Proliferated calli
were survived (100%) for one day only on all basta
level. After one day of shifting on basta selection
pressure, basta showed effect. It was observed that at
1 mg/L and 3 mg/L calli survived (80%) while at 5 mg/L,
7 mg/L and 10 mg/L calli survival response was 50%,
40% and 30% respectively. On fifth day, survival
frequency of embryo was 5% and 3% on 5 mg/L, 7 mg/L,
and 10 mg/L basta level respectively. While complete
death of embryo was observed at 5 mg/L, 7 mg/L, and
10 mg/L basta level on 7
th
day (Figure 3 and Figure 4).
Figure 3 Optimization of basta dose for selection of transgenic
regenerants in genotype S-2003-us-127
Figure 4 Calli response on regeneration selection medium (RSM) at various levels of basta
Note: A: 1 mg/L; B: 3 mg/L; C: 5mg/L; D: 7mg/L; E: 10 mg/L
But 50% and 45 % embryo were survived at 1 mg/L and 3
mg/L basta respectively. On 1 mg/L basta regeneration
was starting after 12 days (Figure 5).
After the selection of basta doses for genotype
S-2003-us-359 and S-2003-us-127, affectivity or
efficiency of these optimal doses was determined by
integrating the bar gene in to these genotypes using
Figure 5 Regeneration on RSM having 1 mg/L basta