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Cotton Genomics and Genetics 2012, Vol.3, No.1, 1
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Table 4 The incidence of
Fusarium oxysporum
f. sp
vasinfectum
in transgenic and non-transgenic plants after inoculation
Variety
Total The 1
st
day
The 6
th
day
The 11
th
day
The 16
th
day
The 21
th
day
Diseased
plants
Incidenc
e (%)
Diseased
plants
Incidenc
e (%)
Diseased
plants
Incidenc
e (%)
Diseased
plants
Incidence
(%)
Diseased
plants
Incidence
(%)
Transgenic
“Junmian No.1”
128 0
0
16
12.5
28
21.9
32
25.0
48
37.5
Transgenic
“Zhong35”
153 0
0
17
11.1
23
15.0
34
22.2
34
22.2
Non-transgenic
“Junmian No.1”
150 0
0
33
22.0
84
56.0
100
66.7
100
66.7
Non-transgenic
“Zhong35”
150 0
0
15
10.0
45
30.0
60
40.0
75
50.0
Figure 5 Symptom of transgenic and non-transgenic plants after
inoculation in 21days
Note: A: Transgenic “Zhong35”; B: Transgenic “Junmian 1
hao”; C: Non-transgenic “Zhong35”; D: Non-transgenic
“Junmian 1 hao”
regeneration derived from the apical meristem (Sun et
al., 2009; Zhao 2009; Weng et al., 2009; Zhou et al.,
2009; Lv et al., 2004; Balasubrmani ea al., 2003).
In this study, we adopted Agrobacterium-mediated
approach to transform cotton shoot apex explant by
modifying the growth period, the infection time,
pre-culture time, which greatly improved the
efficiency of genetic transformation.
PCR detection for the target gene and herbicide
marker gene and RT-PCR technique for the target gene
verified that the exogenous target gene has been
integrated into the cotton genome in the level of
integration and expression.
In this study, the strain of Xinjiang cotton Fusarium
wilt is a strong virulence strain, transgenic
SNC1
cotton varieties were significantly lower incidence
than that of non-transgenic controls, indicating that
transgenic
SNC1
cotton acquired cotton Fusarium wilt
resistance. However, this was the indoor results of T
1
generation plants, yet remaining to verify the
advanced generation of transgenic cotton.
3 Materials and Methods
3.1 Receptor materials
The receptor materials were the South Xinjiang main
cultivated upland varieties, Zhong35 and Juanmian 1
hao, which provided by Key Lab of Crops Cell
Engineering in the Institute of Nuclear Biotech of
Xinjiang Academy of Agricultural Sciences.
3.2 Plasmids and
Agrobacterium
strains
Agrobacterium
strain GV3101 containing
SNC1
gene
was provided by Dr. Li Xin from University of
British Columbia, Canada. Strain was selected with
kanamycin 50 mg/L and rifampicin 50 mg/L, and the
herbicide as selection marker used for the resistance of
regeneration plants.
3.3 Reagents
Conventional reagents were domestic analytical grade.
Reverse transcriptases were purchased from Promega
Corporation. 10×Bufffer, dNTPs and
Taq
enzymes
purchased from Shanghai Biological Engineering
Technology Co., Ltd..; the SNC1 primers for PCR
detection were synthesized by the Beijing Huada