Heterologous overexpression of StERF3 triggers cell death in Nicotiana benthamiana

Programmed cell death plays a crucial role in plant development and disease defense. Here, we report that the expression of StERF3, a potato EAR motif-containing transcription factor, promotes Phytophthora infestans colonization in Nicotiana benthamiana. Transient overexpression of StERF3 induces cell death in N. benthamiana leaves. The substitution of two key amino acids (14th and 19th) in its ERF domain (the DNA binding domain) dramatically altered its cell death-inducing ability. In addition, StERF3(Delta EAR) EAR motif-deletion or StERF3(AAA )mutation abolished the cell death-inducing ability. StERF3 interacted with the co-repressors Topless-related protein 1 (StTPL1) and Topless-related protein 3 (StTPL3) via the EAR motif. Moreover, cell death induced by StERF3 was facilitated by co-expression with StTPL1 or StTPL3. Virus-induced gene silencing (VIGS) of NbTPL1 and NbTPL3 in N. benthamiana compromised the cell death-inducing ability of StERF3. Furthermore, StERF3-induced cell death accompanied with ROS bursts and the upregulation of the respiratory burst oxidase homolog (Rboh) genes NbRbohA and NbRbohC. In addition, several cell death regulator genes, including NbCRTD, NbNCBP, and NbBCPL, and a hypersensitive cell death marker gene Hin1 were upregulated. StERF3 may positively regulate cell death through its EAR motif-mediated transcriptional repressor activity by inhibiting the expression of genes potentially coding the repressor of cell death (CD).

Automated summary

The potato transcription factor StERF3 promotes colonization of Phytophthora infestans in Nicotiana benthamiana and induces cell death. Its cell death-inducing ability is affected by two key amino acids in the ERF domain, and deletion or mutation of the EAR motif diminishes this ability. StERF3 interacts with co-repressors via the EAR motif and facilitates cell death, which is compromised by silencing of related genes in Nicotiana benthamiana. Moreover, StERF3-induced cell death is associated with ROS bursts and upregulation of cell death regulator genes.