Basic HTML Version
Cotton Genomics and Genetics 2012, Vol.3, No.1, 1
-
7
http://cgg.sophiapublisher.com
3
Table 2 Effect of infection time on the growth status of shoot apex
Infection time (min) Variety
Average No. of
transformed plants
Average No. of
survival plants
Transformation rate (%)
10
Zhong35
132
74
56.1
Junmian No.1
108
67
62.0
20
Zhong35
120
92
76.7
Junmian No.1
118
88
74.6
30
Zhong35
122
59
48.4
Junmian No.1
98
51
52.0
Table 3 Effects of co-culture time on the growth status of shoot apex
Co-culture time (d) Variety
Average No. of
transformed plants
Average No. of
survival plants
Transformation rate (%)
1
Zhong35
96
55
57.3
Junmian No.1
86
50
58.1
2
Zhong35
136
90
66.2
Junmian No.1
121
87
71.9
3
Zhong35
132
61
46.2
Junmian No.1
124
60
48.4
4
Zhong35
105
47
44.8
Junmian No.1
112
49
43.8
1.4 Obtaining transgenic
SNC1
cotton
The transgenic cotton seedlings being rooting were
planted in the flower pots that filled with the steriled
potting loam and vermiculite (mixed by 1:1) and
irrigated nutrient solution prior to bagging, and placed
in the incubator to continuing foster. Then after 10
days culture the robust transgenic cotton seedlings
transplanted into the greenhouse (Figure 1) from out
of the pots. The results showed that the 45 individuals
of putative transgenic resistant plants were obtained,
and the 16 individuals were proved to be transgenic
SNC1 gene positive cotton plants by PCR assays
(Figure 2). Transgenic cotton seeds were harvested
respectively according to the varieties.
1.5 PCR detection of transgenic
SNC1
cotton
plants in T
1
generation
Let all T
0
generation seeds of transgenic plant seeds
continue to grow, when the cotton grew up in the
five-leaf stage, genomic DNA of T
1
generation cotton
plant was extracted form leaves, PCR detection for the
target gene and selectable marker Bar gene were
carried out.
The PCR product of the target gene
SNC1
by the
electrophoresis showed that 281 plants of the detected
860 cotton plants had a single clear band with 790 bp
in length that located in the same position as the
plasmid amplification products; while there was no band
Figure 1 Transgenic cotton at different growth stages
Note: A: Shoot apex; B: Regenerated plantlets; C: Rooted
plantlet; D: Transgenic plantlets planted in soil