Basic HTML Version
盛德鹏等
, 2011,
蛋白激发子
PeaT1
转化水稻恢复系多系一号的研究
,
分子植物育种
Vol.9 No.61 (doi: 10.5376/mpb.cn.2011.09.0061)
1455
根据
peaT1
基因序列设计的特异引物
4 (
表
2)
，反应
条件为
94
℃预变性
5 min
，
94
℃
30 s
，
59
℃
30 s
，
72
℃
1 min 30 s
，
35
个循环，最后
72
℃延伸
10 min
。
DNA
分子杂交按照罗氏地高辛检测试剂盒
(
Ⅱ
)
说
明书的方法。
3.5.3
转基因植株的
Western blot
检测
选取
Southern
检测阳性的
T
2
代转基因植株叶片
提取总蛋白，参照
Coca
等的方法进行
Western blot
杂交鉴定
(Coca et a1., 2006)
。抗体以本实验室从极
细链格孢菌中纯化的蛋白激发子
PeaT1
为抗原制备。
作者贡献
盛德鹏、玉山江、买买提和郭丽华是本研究的实验设
计和实验研究的执行人；盛德鹏、郭丽华和曾红梅完成数
据分析，论文初稿的写作；曾红梅、杨秀芬和袁京京参与
实验设计，试验结果分析；邱德文是项目的构思者及负责
人，指导实验设计，论文写作和修改。全体作者都阅读并
同意最终的文本。
致谢
本 研 究 由 转 基 因 生 物 新 品 种 培 育 重 大 专 项
(2009ZX08001
-
018B)
资助。感谢中国水稻研究所黄世文副
研究员赠送水稻种子和在实验中提供的帮助。
参考文献
Chan M.T., Lee. T.M., and Chang H.H., 1992, Transformation
of Indica rice (
Oryza sativa
L.) mediated by
Agrobacterium
tumefaciens
, Plant Cell Physiology, 33(5): 577-583
Coca M., Peñas G., Gómez J., Campo S., Bortolotti C.,
Messeguer J., and Segundo B.S., 2006, Enhanced
resistance to the rice blast fungus Magnaporthe grisea
conferred by expression of a cecropin A gene in transgenic
rice, Planta, 223(3): 392-406
Hiei Y., Komari T., Kubo T., and Tomoaki K., 1997,
Transformation of rice mediated by
Agrobacterium
tumefaciens
, Plant molecular Biology, 35(1-2): 205-218
Hiei Y., Ohta S., Komari T., and Kumashiro T., 1994, Efficient
transformation of rice (
Oryza sativa
L.) mediated by
Agrobacterium and sequence analysis of the boundaries of
the T-DNA, Plant Journal., 6(2): 271-282
Kenjirou Ozawa., 2010, Establishment of a high efficiency
Agrobacterium
-mediated transformation system of rice
(
Oryza sativa
L.), Plant Science, 176(4): 522-527
Liu S., Cheng H., Wang F., and Zhu Y., 2002, DNA
polymorphism of main restorer lines of hybrid rice in
China, Zhongguo Shuidao Kexue (Chinese Journal of Rice
Science), 16(1): 1-5 (
刘殊
,
程慧
,
王飞
,
朱英国
, 2002,
我国杂交水稻主要恢复系的
DNA
多态性研究
,
中国水
稻科学
, 16(1): 1-5
Mao J., Liu Q., Yang X., Long C., Zhao M., Zeng H., Liu H.,
Yuan J., and Qiu D., 2010, Purification and expression of a
protein elicitor from Alternaria tenuissima and
elicitor-mediated defense responses in tobacco, Annals of
Applied Biology, 156(3): 411-420
Qiu D., Mao J., Yang X., and Zeng H., 2009, Expression of an
elicitor-encoding gene from Magnaporthe grisea enhances
resistance against blast disease in transgenic rice, Plant
Cell Reports, 28(6): 925-933
Saharan V., Yadav R.C., Yadav N.R., and Ram K., 2004, Studies
on improved
Agrobacterium
-mediated transformation in two
indica rice (
Oryza sativa
L.), African Journal of
Biotechnology, 3(11): 572-575
Shao M., and Wang J.S., 2004, Transformation of rice with
hrfAxoo
gene and resistance of the transgenic plant to
bacterial leaf blight, Nanjing Nongye Daxue Xuebao
(Journal of Nanjing Agricultural University), 27(4): 36-40
(
邵敏
,
王金生
, 2004,
转
HrfAXoo
基因水稻对白叶枯病
的抗性
,
南京农业大学学报
, 27(4): 36-40)
Toki S., Hara N., Ono K., Onodera H., Tagiri A., Oka S., and
Tanaka H., 2006, Early infection of scutellum tissue with
Agrobacterium allows high-speed transformation of rice,
The Plant Journal., 47(6): 969-976
Upadhyaya N.M., Surin B., Ramm K., Gaudron J., Schünmann
P.H.D., Taylor W., Waterhouse P.M., and Wang M.B., 2000,
Agrobacterium
-mediated transformation of Australian rice
cultivars Jarrah and Amaroo using modified promoters and
selectable markers, Australian Journal of Plant Physiology,
27(3): 201-210
Yang X.F., Qiu D.W., Zeng H.M., Yuan J.J., Mao J.J, 2009a,
Purification and characterization of a glycoprotein elicitor
from
Alternaria tenuissima
, World Journal of Microbiology
and Biotechnology, 25(11): 2035-2042
Yimian M., Luo L., Chengguang Z., Changhui S., Bo X., Jun F.,
Jiuyou T., Anding L., Shouyun C., Gupo L., Qian Q.,
Yongbiao X., and Chengcai C., 2009, Molecular analysis
of rice plants harboring a multi-functional T-DNA tagging
system, Journal of Genetics and Genomics, 36(5): 267-276