Transformation of potato (Solanum tuberosum) using particle bombardment

被引:21
作者
Romano, A
Raemakers, K
Visser, R
Mooibroek, H
机构
[1] Univ Wageningen & Res Ctr, Agrotechnol Res Inst, Wageningen UR ATO, NL-6700 AA Wageningen, Netherlands
[2] Univ Wageningen & Res Ctr, Lab Plant Breeding, NL-6700 AY Wageningen, Netherlands
关键词
Solanum tuberosum; internodes; particle bombardment; regeneration; stable transformation;
D O I
10.1007/s002990000314
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Internodes, leaves and tuber slices from potato (Solanum tuberosum), genotype 1024-2, were subjected to particle bombardment. Transient expression was optimized using the uidA and the luc reporter genes that encode beta -glucuronidase (GUS) and luciferase, respectively. Stable transformation was achieved using the neomycin phosphotransferase (nptII) gene, which confers resistance to the antibiotic kanamycin. The influence of biological parameters (tissue type, growth period before bombardment, pre- and post-bombardment osmoticum treatment) and physical parameters (helium pressure, tissue distance) that are known to possibly affect stable transformation were investigated. Putative transgenic plants, which rooted in media containing kanamycin, were obtained from all of the tissues tested although there were large differences in the efficiency: internodes (0.77 plants per bombarded explant), microtuber slices (0.10 plants per bombarded explant) and leaves (0.02 plants per bombarded explant). Southern blot analysis of putative transgenic plants confirmed the integration of the transgenes into plant DNA. The results indicate that an efficient particle bombardment protocol is now available for both transient and stable transformation of potato internodal segments, thus contributing to an enhanced flexibility in the delivery of transgenes to this important food crop.
引用
收藏
页码:198 / 204
页数:7
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