Agrobacterium-mediated floral dip transformation of the model polyploid species Arabidopsis kamchatica
Polyploidization has played an important role in the speciation and diversification of plant species. However, genetic analyses of polyploids are challenging because the vast majority of the model species are diploids. The allotetraploid Arabidopsis kamchatica, which originated through the hybridization of the diploid Arabidopsis halleri and Arabidopsis lyrata, is an emerging model system for studying various aspects of polyploidy. However, a transgenic method that allows the insertion of a gene of interest into A. kamchatica is still lacking. In this study, we investigated the early development of pistils in A. kamchatica and confirmed the formation of open pistils in young flower buds (stages 8–9), which is important for allowing Agrobacterium to access female reproductive tissues. We established a simple Agrobacterium-mediated floral dip transformation method to transform a gene of interest into A. kamchatica by dipping A. kamchatica inflorescences bearing many young flower buds into a 5% sucrose solution containing 0.05% Silwet L-77 and Agrobacterium harboring the gene of interest. We showed that a screenable marker comprising fluorescence-accumulating seed technology with green fluorescent protein was useful for screening the transgenic seeds of two accessions of A. kamchatica subsp. kamchatica and an accession of A. kamchatica subsp. kawasakiana.
KeywordsAgrobacterium-mediated floral dip Allopolyploid Arabidopsis kamchatica Tetraploid Transformation Transgenic plant
We would like to thank Misako Yamazaki, Reiko Akiyama, Yuanyuan Huang, and Rie Shimizu-Inatsugi for technical support and discussion. This research was supported by the Swiss National Science Foundation, SystemsX.ch, JST CREST Grant Number JPMJCR16O3, Japan, and a KAKENHI Grant (nos. 16H06469, 16H06464, 16K21727).
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Conflict of interest
The authors declare that they have no conflicts of interest.
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