Alternative selectable markers for potato transformation using minimal T-DNA vectors

被引:25
作者
Barrell, PJ
Shang, YJ
Cooper, PA
Conner, AJ
机构
[1] New Zealand Inst Crop & Food Res Ltd, Christchurch, New Zealand
[2] Lincoln Univ, Soil Plant & Ecol Sci Div, Canterbury, New Zealand
关键词
binary vectors; plant transformation; potato; selectable markers; Solanum tuberosum; T-DNA vectors;
D O I
10.1023/A:1016013426923
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Alternative selection systems for plant transformation are especially valuable in clonal crops, such as potato (Solanum tuberosum L.), to pyramid transgenes into the same cultivar by successive transformation events. We have modified the pGPTV series of binary vectors to construct pMOA1 to pMOA5, resulting in a series of essentially identical binary vectors except for the presence of different selectable marker genes. These selectable marker genes are tightly inserted between the left and right T-DNA borders and confer resistance to kanamycin (nptII), hygromycin (hpt), methotrexate (dhfr), phosphinothricin (bar), or phleomycin (ble). The T-DNA of all the vectors is based on the minimal features necessary for plant transformation, with no extraneous DNA segments that may be unacceptable to regulatory authorities for general release of transgenic plants. A series of unique restriction sites exists between the right border and each selectable marker gene for subsequent insertion of useful genes. We have also developed improved culture procedures for potato transformation and used the pMOA1 to pMOA5 binary vectors to define stringent selection conditions for each marker gene. Combining these advances improved the frequency of recovering transformed potato plants while maintaining a low frequency of escapes. The relative efficiency of recovering transgenic potato lines with each selectable marker gene can be summarised as: kanamycin resistance>hygromycin resistance>phosphinothricin resistance>phleomycin resistance>methotrexate resistance.
引用
收藏
页码:61 / 68
页数:8
相关论文
共 23 条
[1]   NEW PLANT BINARY VECTORS WITH SELECTABLE MARKERS LOCATED PROXIMAL TO THE LEFT T-DNA BORDER [J].
BECKER, D ;
KEMPER, E ;
SCHELL, J ;
MASTERSON, R .
PLANT MOLECULAR BIOLOGY, 1992, 20 (06) :1195-1197
[2]   BINARY AGROBACTERIUM VECTORS FOR PLANT TRANSFORMATION [J].
BEVAN, M .
NUCLEIC ACIDS RESEARCH, 1984, 12 (22) :8711-8721
[3]  
CONNER A J, 1986, New Zealand Journal of Technology, V2, P83
[4]   Genetic engineering of crops as potential source of genetic hazard in the human diet [J].
Conner, AJ ;
Jacobs, JME .
MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS, 1999, 443 (1-2) :223-234
[5]  
CONNER AJ, 1991, NEW ZEAL J CROP HORT, V19, P1
[6]   PLANT-BREEDING AND SEED MARKETING OPTIONS FOR THE INTRODUCTION OF TRANSGENIC INSECT-RESISTANT CROPS [J].
CONNER, AJ ;
CHRISTEY, MC .
BIOCONTROL SCIENCE AND TECHNOLOGY, 1994, 4 (04) :463-473
[7]  
CONNER AJ, 1997, COMMERCIALISATION TR, P23
[8]   Localization of Ds-transposon containing T-DNA inserts in the diploid transgenic potato: Linkage to the R1 resistance gene against Phytophthora infestans (Mont) de Bary [J].
ElKharbotly, A ;
Jacobs, JME ;
Hekkert, BTL ;
Jacobsen, E ;
Ramanna, MS ;
Stiekema, WJ ;
Pereira, A .
GENOME, 1996, 39 (02) :249-257
[9]   Positive selection: a plant selection principle based on xylose isomerase, an enzyme used in the food industry [J].
Haldrup, A ;
Petersen, SG ;
Okkels, FT .
PLANT CELL REPORTS, 1998, 18 (1-2) :76-81
[10]   The xylose isomerase gene from Thermoanaerobacterium thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent [J].
Haldrup, A ;
Petersen, SG ;
Okkels, FT .
PLANT MOLECULAR BIOLOGY, 1998, 37 (02) :287-296