Cut-dip-budding delivery system enables genetic modifications in plants without tissue culture

Of the more than 370000 species of higher plants in nature,fewer than 0.1%can be geneticallymodified due to limitations of the current gene delivery systems.Even for those that can be genetically modified,the modification involves a tedious and costly tissue culture process.Here,we describe an extremely simple cut-dip-budding(CDB)delivery system,which uses Agrobacterium rhizogene to inoculate explants,generating transformed roots that produce transformed buds due to root suckering.We have successfully used CDB to achieve the heritable transformation of plant species inmultiple plant families,including two herbaceous plants(Taraxacum kok-saghyz and Coronilla varia),a tuberous root plant(sweet potato),and three woody plant species(Ailanthus altissima,Aralia elata,and Clerodendrum chinense).These plants have previously been difficult or impossible to transform,but the CDB method enabled efficient transformation or gene editing in them using a very simple explant dipping protocol,under non-sterile conditions and without the need for tissue culture.Our work suggests that large numbers of plants could be amenable to genetic modifications using the CDB method.