作者：Wu, H.， Acanda, Y.， Jia, H. G.， Wang, N. A.， Zale, J.
作者单位：[Wu, Hao; Acanda, Yosvanis; Jia, Hongge; Wang, Nian; Zale, Janice] Univ Florida, Inst Food & Agr Sci, Dept Plant Pathol, Citrus Res & Educ Ctr, Lake Alfred, FL 33850 USA. Wu, H; Zale, J (reprint author), Univ Florida, Inst Food & Agr Sci,
摘要：The development of transgenic citrus plants by the biolistic method. A protocol for the biolistic transformation of epicotyl explants and transgenic shoot regeneration of immature citrange rootstock, cv. Carrizo (Citrus sinensis Osb. x Poncirus trifoliata L. Raf.) and plant regeneration is described. Immature epicotyl explants were bombarded with a vector containing the nptII selectable marker and the gfp reporter. The number of independent, stably transformed tissues/total number of explants, recorded by monitoring GFP fluorescence 4 weeks after bombardment was substantial at 18.4 %, and some fluorescing tissues regenerated into shoots. Fluorescing GFP, putative transgenic shoots were micro-grafted onto immature Carrizo rootstocks in vitro, confirmed by PCR amplification of nptII and gfp coding regions, followed by secondary grafting onto older rootstocks grown in soil. Southern blot analysis indicated that all the fluorescing shoots were transgenic. Multiple and single copies of nptII integrations were confirmed in five regenerated transgenic lines. There is potential to develop a higher throughput biolistics transformation system by optimizing the tissue culture medium to improve shoot regeneration and narrowing the window for plant sampling. This system will be appropriate for transformation with minimal cassettes.
关键词：Citrus transgenics, Carrizo citrange, Biolistics, Particle bombardment, Genetic
文献注录：Wu, H.， Acanda, Y.， Jia, H. G.， Wang, N. A.， Zale, J.. Biolistic transformation of Carrizo citrange (Citrus sinensis Osb. x Poncirus trifoliata L. Raf.) [J]. Plant Cell Reports. 2016, 35(9): 1955-1962.
报/刊名：《Plant Cell Reports》，发表于2016 / 第 35(9) 期。
文献类型： [J]
页码：第 1955-1962 页 / 共8 页