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

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

作者机构：Beijing Key Laboratory of Agricultural Genetic Resources and BiotechnologyInstitute of BiotechnologyBeijing Academy of Agriculture and Forestry SciencesBeijing 100097China College of Life SciencesCapital Normal UniversityBeijing 100048China 

出 版 物：《Journal of Integrative Plant Biology》 (植物学报（英文版）)

年 卷 期：2022年第64卷第6期

页      面：1145-1156页

核心收录：

学科分类：09[农学] 0901[农学-作物学] 

基　　金：funded by Innovation Program of Beijing Academy of Agriculture and Forestry Sciences (KJCX20200407, KJCX20200204, KJCX20200205) Beijing Postdoctoral Research Foundation (2018-ZZ-062) Beijing Academy of Agriculture and Forestry Sciences Postdoctoral Research Foundation (2018ZZ-004) the Program of Beijing Municipal Commission of Science and Technology (Z171100001517001) 

主　　题：exogenous genes genotype independence magnetic nanoparticles maize inbred lines pollen transfection pollen aperture 

摘      要：Current gene delivery methods for maize are limited to specific genotypes and depend on timeconsuming and labor-intensive tissue culture ***,we report a new method to transfect maize that is culture-free and genotype *** 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),β-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 *** fluorescence was detected in 22%transfected pollen grains,and GUS stained 55%embryos at 18 d after *** fluorescence was detected in both silk filaments and immature *** 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 ***,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 *** 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.