The C‐terminal half of Phytophthora infestans RXLR effector AVR3a is sufficient to trigger R3a‐mediated hypersensitivity and suppress INF1‐induced cell death in Nicotiana benthamiana

Abstract

Summary: The RXLR cytoplasmic effector AVR3a of Phytophthora infestans confers avirulence on potato plants carrying the R3a gene. Two alleles of Avr3a encode secreted proteins that differ in only three amino acid residues, two of which are in the mature protein. Avirulent isolates carry the Avr3a allele, which encodes AVR3a KI (containing amino acids C 19, K 80 and I 103), whereas virulent isolates express only the virulence allele avr3a, encoding AVR3a EM (S 19, E 80 and M 103). Only the AVR3a KI protein is recognized inside the plant cytoplasm where it triggers R3a‐mediated hypersensitivity. Similar to other oomycete avirulence proteins, AVR3a KI carries a signal peptide followed by a conserved motif centered on the consensus RXLR sequence that is functionally similar to a host cell‐targeting signal of malaria parasites. The interaction between Avr3a and R3a can be reconstructed by their transient co‐expression in Nicotiana benthamiana. We exploited the N. benthamiana experimental system to further characterize the Avr3a–R3a interaction. R3a activation by AVR3a KI is dependent on the ubiquitin ligase‐associated protein SGT1 and heat‐shock protein HSP90. The AVR3a KI and AVR3a EM proteins are equally stable in planta, suggesting that the difference in R3a‐mediated death cannot be attributed to AVR3a EM protein instability. AVR3a KI is able to suppress cell death induced by the elicitin INF1 of P. infestans, suggesting a possible virulence function for this protein. Structure–function experiments indicated that the 75‐amino acid C‐terminal half of AVR3a KI, which excludes the RXLR region, is sufficient for avirulence and suppression functions, consistent with the view that the N‐terminal region of AVR3a KI and other RXLR effectors is involved in secretion and targeting but is not required for effector activity. We also found that both polymorphic amino acids, K 80 and I 103, of mature AVR3a contribute to the effector functions.