Efficient and genotype independent maize transformation using pollen transfected by DNA-coated magnetic nanoparticles

文献类型： 外文期刊

第一作者： Wang, Zuo-Ping

作者： Wang, Zuo-Ping;Zhang, Zhong-Bao;Zheng, Deng-Yu;Zhang, Tong-Tong;Li, Xiang-Long;Zhang, Chun;Wei, Jian-Hua;Wu, Zhong-Yi;Zhang, Tong-Tong;Yu, Rong

作者机构：

关键词： exogenous genes; genotype independence; magnetic nanoparticles; maize inbred lines; pollen transfection; pollen aperture

期刊名称：JOURNAL OF INTEGRATIVE PLANT BIOLOGY （ 影响因子：9.106； 五年影响因子：8.241 ）

ISSN： 1672-9072

年卷期： 2022 年 64 卷 6 期

页码：

收录情况： SCI

摘要： Current gene delivery methods for maize are limited to specific genotypes and depend on time-consuming and labor-intensive tissue culture techniques. Here, we report a new method to transfect maize that is culture-free and genotype independent. To enhance efficiency of DNA entry and maintain high pollen viability of 32%-55%, transfection was performed at cool temperature using pollen pretreated to open the germination aperture (40%-55%). Magnetic nanoparticles (MNPs) coated with DNA encoding either red fluorescent protein (RFP), beta-glucuronidase gene (GUS), enhanced green fluorescent protein (EGFP) or bialaphos resistance (bar) was delivered into pollen grains, and female florets of maize inbred lines were pollinated. Red fluorescence was detected in 22% transfected pollen grains, and GUS stained 55% embryos at 18 d after pollination. Green fluorescence was detected in both silk filaments and immature kernels. The T1 generation of six inbred lines showed considerable EGFP or GUS transcripts (29%-74%) quantitated by polymerase chain reaction, and 5%-16% of the T1 seedlings showed immunologically active EGFP or GUS protein. Moreover, 1.41% of the bar transfected T1 plants were glufosinate resistant, and heritable bar gene was integrated into the maize genome effectively as verified by DNA hybridization. These results demonstrate that exogenous DNA could be delivered efficiently into elite maize inbred lines recalcitrant to tissue culture-mediated transformation and expressed normally through our genotype-independent pollen transfection system.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序