Haymes K M, Davis T M
Department of Plant Biology, Rudman Hall, University of New Hampshire, Durham, NH 03824, USA Fax no.: +1-603-862-3784 E-mail:
Plant Cell Rep. 1998 Feb;17(4):279-283. doi: 10.1007/s002990050392.
Agrobacterium-mediated transformation was used to stably introduce β-glucuronidase (gus) and neomycin phosphotransferase (nptII) marker genes into Alpine' Fragaria vesca FRA 197, a diploid (2n = 2x = 14) strawberry. R0 generation transformants derived from a single clump of kanamycin-resistant callus were vegetatively propagated. The presence of the gus and nptII genes in five clonal R0 runner plants was confirmed by PCR. Southern analysis suggested two sites of nptII insertion. When R1 generation seedlings obtained via self-fertilization of R0 plants were tested by histochemical assay, 591 were GUS positive and 39 were GUS negative. The R1 segregation data fit a 15 : 1 ratio (0.5 > P > 0.25), consistent with the independent segregation of two transgene insertion loci. These results demonstrate the suitability of Alpine' F. vesca for transgene research in strawberry.
采用农杆菌介导的转化方法，将β-葡萄糖醛酸酶（gus）和新霉素磷酸转移酶（nptII）标记基因稳定导入二倍体（2n = 2x = 14）草莓“高山”野草莓FRA 197中。从一丛卡那霉素抗性愈伤组织获得的R0代转化体进行了无性繁殖。通过PCR证实了五个克隆的R0代匍匐茎植株中存在gus和nptII基因。Southern分析表明nptII有两个插入位点。当通过R0代植株自交获得的R1代幼苗进行组织化学检测时，591株为GUS阳性，39株为GUS阴性。R1代的分离数据符合15:1的比例（0.5 > P > 0.25），这与两个转基因插入位点的独立分离一致。这些结果表明“高山”野草莓适用于草莓的转基因研究。
