An ancient cis‐element targeted by Ralstonia solanacearum TALE‐like effectors facilitates the development of a promoter trap that could confer broad‐spectrum wilt resistance

Abstract

SummaryRalstonia solanacearum, a species complex of bacterial plant pathogens that causes bacterial wilt, comprises four phylotypes that evolved when a founder population was split during the continental drift ~180 million years ago. Each phylotype contains strains with RipTAL proteins structurally related to transcription activator‐like (TAL) effectors from the bacterial pathogen Xanthomonas. RipTALs have evolved in geographically separated phylotypes and therefore differ in sequence and potentially functionality. Earlier work has shown that phylotype I RipTAL Brg11 targets a 17‐nucleotide effector binding element (EBE) and transcriptionally activates the downstream arginine decarboxylase (ADC) gene. The predicted DNA binding preferences of Brg11 and RipTALs from other phylotypes are similar, suggesting that most, if not all, RipTALs target the Brg11‐EBE motif and activate downstream ADC genes. Here we show that not only phylotype I RipTAL Brg11 but also RipTALs from other phylotypes activate host genes when preceded by the Brg11‐EBE motif. Furthermore, we show that Brg11 and RipTALs from other phylotypes induce the same quantitative changes of ADC‐dependent plant metabolites, suggesting that most, if not all, RipTALs induce functionally equivalent changes in host cells. Finally, we report transgenic tobacco lines in which the RipTAL‐binding motif Brg11‐EBE mediates RipTAL‐dependent transcription of the executor‐type resistance (R) gene Bs4C from pepper, thereby conferring resistance to RipTAL‐delivering R. solanacearum strains. Our results suggest that cell death‐inducing executor‐type R genes, preceded by the RipTAL‐binding motif Brg11‐EBE, could be used to genetically engineer broad‐spectrum bacterial wilt resistance in crop plants without any apparent fitness penalty.

Molecular Basis of Plant–Pathogen Interactions in the Agricultural Context

“…However, an improved understanding of the specificity of DNA binding to TALEs provides more insights into developing immunity against various Xanthomonas strains [ 130 , 131 ]. This approach was also employed to engineer RipTALs for resistance development against R. solanacearum in tomatoes [ 132 ].…”