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Molecular Plant Breeding 2011, Vol.2, No.5, 30
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Table 1 Effect of Kan on the root growth of adventitious buds
Rooting status of adventitious buds
Kanamycin dose (mg/L)
Non-transformed adventitious buds
Transgenic adventitious buds
0
Normal rooting and robust growth
All were normal in rooting, grew robustly, roots
were long and more
50
No roots, normal color in branches and leaves
Grew roots partially, with 5~8 roots
75
No roots, stunted plants
Grew roots partially, with 3~4 roots
Table 2 Rooting rate of transformed adventitious buds
Selection
No. of adventitious buds inoculated
No. of adventitious buds rooted
Rooting rate
First selection
38
29
76.32%
Second selection
29
25
86.21%
Figure 1 Effect of different Kan concentrations on the root
growth of adventitious buds
1.3 PCR detection
The PCR positive rates of the transformant lines and
plants were 93.75% and 61.15% respectively (Table 3).
Results of PCR electrophoresis of partial transformant
plants are shown in Figure 4. Both positive plants and
positive
Agrobacterium
could be amplified with a
band of approximately 720 bp, while the
non-transformed plants had no target band (Figure 2).
1.4 Southern Blot analysis
The lines and plants detected positive by PCR assay
were selected for Southern Blot assay. Of the 13 plants
detected, seven showed positive response, yielding a
rate of 53.85% (Figure 3), which indicates that the
target gene had been successfully transformed into the
potato genome.
Figure 2 PCR analysis of individual
hrap-
transgenic plants of
potatoes
Note: 1~7: transgenic plants; 8: Negative control (non-transgenic
plant); 9: Positive control, PCR of bacterium liquid; Marker:
D2000
Table 3 PCR analysis of transgenic lines and plants
No. of lines
tested
No. of positive
lines
Rate of positive
lines
No. of plants
tested
No. of positive
plants
Rate of positive
plants
16
15
93.75%
157
96
61.15%