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An efficient system composed of maize protoplast transfection and HPLC-MS for studying the biosynthesis and regulation of maize benzoxazinoids
Gao, Lei1,2; Shen, Guojing1; Zhang, Lingdan1; Qi, Jinfeng1; Zhang, Cuiping1; Ma, Canrong1; Li, Jing1; Wang, Lei1; Ul Malook, Saif1; Wu, Jianqiang1
Corresponding AuthorWu, Jianqiang(wujianqiang@mail.kib.ac.cn)
2019-11-28
Source PublicationPLANT METHODS
Volume15Issue:1Pages:13
AbstractBackground Insect herbivory poses a major threat to maize. Benzoxazinoids are important anti-insect secondary metabolites in maize, whose biosynthetic pathway has been extensively studied. However, yet little is known about how benzoxazinoids are regulated in maize, partly due to lack of mutant resources and recalcitrance to genetic transformation. Transient systems based on mesophyll- or cultured cell-derived protoplasts have been exploited in several plant species and have become a powerful tool for rapid or high-throughput assays of gene functions. Nevertheless, these systems have not been exploited to study the regulation of secondary metabolites. Results A protocol for isolation of protoplasts from etiolated maize seedlings and efficient transfection was optimized. Furthermore, a 10-min-run-time and highly sensitive HPLC-MS method was established to rapidly detect and quantify maize benzoxazinoids. Coupling maize protoplast transfection and HPLC-MS, we screened a few genes potentially regulating benzoxazinoid biosynthesis using overexpression or silencing by artificial microRNA technology. Conclusions Combining the power of maize protoplast transfection and HPLC-MS analysis, this method allows rapid screening for the regulatory and biosynthetic genes of maize benzoxazinoids in protoplasts, before the candidates are selected for in planta functional analyses. This method can also be applied to study the biosynthesis and regulation of other secondary metabolites in maize and secondary metabolites in other plant species, including those not amenable to transformation.
KeywordMaize Benzoxazinoids Protoplasts Transient gene expression Transcription factor HPLC-MS
DOI10.1186/s13007-019-0529-2
Indexed BySCI ; SCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Plant Sciences
WOS SubjectBiochemical Research Methods ; Plant Sciences
WOS IDWOS:000499894000001
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Document Type期刊论文
Identifierhttp://ir.kib.ac.cn/handle/151853/70067
Collection资源植物与生物技术所级重点实验室
Corresponding AuthorWu, Jianqiang
Affiliation1.Chinese Acad Sci, Dept Econ Plants & Biotechnol, Yunnan Key Lab Wild Plant Resources, Kunming Inst Bot, Kunming 650201, Yunnan, Peoples R China
2.Yunnan Univ, Sch Biol Sci, Kunming 650091, Yunnan, Peoples R China
Recommended Citation
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Gao, Lei,Shen, Guojing,Zhang, Lingdan,et al. An efficient system composed of maize protoplast transfection and HPLC-MS for studying the biosynthesis and regulation of maize benzoxazinoids[J]. PLANT METHODS,2019,15(1):13.
APA Gao, Lei.,Shen, Guojing.,Zhang, Lingdan.,Qi, Jinfeng.,Zhang, Cuiping.,...&Wu, Jianqiang.(2019).An efficient system composed of maize protoplast transfection and HPLC-MS for studying the biosynthesis and regulation of maize benzoxazinoids.PLANT METHODS,15(1),13.
MLA Gao, Lei,et al."An efficient system composed of maize protoplast transfection and HPLC-MS for studying the biosynthesis and regulation of maize benzoxazinoids".PLANT METHODS 15.1(2019):13.
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