Development of insect-resistant transgenic cauliflower plants expressing the trypsin inhibitor gene isolated from local sweet potato

被引:50
作者
Ding, LC
Hu, CY [1 ]
Yeh, KW
Wang, PJ
机构
[1] William Paterson Univ, Ctr Appl Sci, Wayne, NJ 07470 USA
[2] Natl Chung Hsing Univ, Grad Inst Agr Biotechnol, Taichung 40227, Taiwan
[3] Natl Taiwan Univ, Dept Bot, Taipei 10764, Taiwan
关键词
cauliflower; Brassica oleracea var. botrytis L; trypsin inhibitor; insect resistance; Agrobacterium mediated transformation;
D O I
10.1007/s002990050497
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Agrobacterium-mediated transformation was used to introduce a trypsin inhibitor gene into Taiwan cauliflower (Brassica oleracea var, botrytis L.) cultivars. The TI gene was isolated from a well-adapted Taiwan sweet potato cultivar and was expected to be especially effective in combating local pests. In vitro regeneration studies indicated that 4-day-old cauliflower seedling hypocotyl segments, pretreated with 2,4-dichlorophenoxyacetic acid for 3 days and incubated on a silver-ion-containing shoot induction medium, gave regeneration rates greater than 95%. Optimum transformation conditions were determined. G418 selection at 15 mg/l was initiated 1 week after cocultivation, and the dose was doubled 1 week later. Over 100 putative transgenic plants were produced. Transgenic status was confirmed by in vitro TI activity, and Southern and Western hybridization assays. The transgenic plants demonstrated in planta resistance to local insects to which the control plants were vulnerable.
引用
收藏
页码:854 / 860
页数:7
相关论文
共 21 条
[1]  
BURNETT L, 1994, PLANT CELL TISS ORG, V37, P253
[2]  
CHI GL, 1990, PLANT CELL REP, V9, P195, DOI 10.1007/BF00232178
[3]   GENETIC-TRANSFORMATION OF CAULIFLOWER (BRASSICA-OLERACEA L VAR BOTRYTIS) BY AGROBACTERIUM-RHIZOGENES [J].
DAVID, C ;
TEMPE, J .
PLANT CELL REPORTS, 1988, 7 (02) :88-91
[4]   TRANSFORMATION OF BRASSICA-NAPUS AND BRASSICA-OLERACEA USING AGROBACTERIUM-TUMEFACIENS AND THE EXPRESSION OF THE BAR AND NEO GENES IN THE TRANSGENIC PLANTS [J].
DEBLOCK, M ;
DEBROUWER, D ;
TENNING, P .
PLANT PHYSIOLOGY, 1989, 91 (02) :694-701
[5]  
Dellaporta S. L., 1985, Molecular Biology of Plants, P36
[6]  
DUAN X, 1996, NAT BIOTECHNOL, V14, P484
[7]   TRANSFORMATION OF CAULIFLOWER (BRASSICA-OLERACEA L VAR BOTRYTIS) - AN EXPERIMENTAL SURVEY [J].
EIMERT, K ;
SIEGEMUND, F .
PLANT MOLECULAR BIOLOGY, 1992, 19 (03) :485-490
[8]   A TECHNIQUE FOR RADIOLABELING DNA RESTRICTION ENDONUCLEASE FRAGMENTS TO HIGH SPECIFIC ACTIVITY [J].
FEINBERG, AP ;
VOGELSTEIN, B .
ANALYTICAL BIOCHEMISTRY, 1983, 132 (01) :6-13
[9]   NUTRIENT REQUIREMENTS OF SUSPENSION CULTURES OF SOYBEAN ROOT CELLS [J].
GAMBORG, OL ;
MILLER, RA ;
OJIMA, K .
EXPERIMENTAL CELL RESEARCH, 1968, 50 (01) :151-+
[10]  
Geiger R., 1986, Methods of Enzymatic Analysis, P119