Vasil Indra K, Vasil Vimla
Laboratory of Plant Cell and Molecular Biology, University of Florida, Gainesville, USA.
Methods Mol Biol. 2006;318:273-83. doi: 10.1385/1-59259-959-1:273.
The protocol described in this chapter was successfully used to produce the first transgenic plants of wheat. It involves high velocity bombardment of explants with DNA-coated microprojectiles (the biolistics procedure). It highlights the importance of selecting the right explants (immature embryos and/or embryogenic callus tissues), pre- and post-osmotic treatment, obtaining high levels of gene expression with the maize ubiquitin promoter, and stringent selection (with bar) during culture to obtain stably transformed and normal fertile plants. The protocol is used widely for genetic transformation of wheat.
本章所述的实验方案已成功用于培育出首批转基因小麦植株。它涉及用包被有DNA的微弹对外植体进行高速轰击(生物弹道技术)。该方案突出了选择合适外植体(未成熟胚和/或胚性愈伤组织)、渗透处理前后处理、使用玉米泛素启动子获得高水平基因表达以及在培养过程中进行严格筛选(使用bar基因)以获得稳定转化且正常可育植株的重要性。该实验方案被广泛用于小麦的遗传转化。