The mutant STAY-GREEN (Cssgr) in cucumber interacts with the CSEP30 protein to elicit a defense response against Podosphaera xanthii

Abstract

Disease-resistant plants activate immune responses by specifically recognition Candidate Secreted Effector Proteins (CSEPs) through resistance (R) proteins. In research on cucumber powdery mildew resistance breeding, several R genes and CSEPs have been identified; however, the specific interactions between R proteins and CSEPs are still largely unexplored. In this study, we used a luciferase reporter assay to identify six CSEPs from Podosphaera xanthii that potentially induce cell death in cucumber. Subsequent yeast two-hybrid analysis revealed that only the mature form of CSEP30 (CSEP30^∆SP) interacted with the cucumber mutant STAY-GREEN (Cssgr), a gene previously recognized for its broad-spectrum resistance in genetic studies. This interaction was confirmed using pull-down and co-immunoprecipitation assays. Additionally, to determine if the interaction leads to phenotypic changes, Cssgr and CSEP30^∆SP were transiently expressed in tobacco leaves. The infiltration of Cssgr in tobacco resulted in reduced chlorosis compared to the wild-type CsSGR. Co-infiltration of Cssgr with CSEP30^∆SP induced distinct dry necrotic lesions, contrasting the effects observed when Cssgr and CSEP30^∆SP were infiltrated separately. Additionally, after P. xanthii infection in moderately powdery mildew-resistant Gy14 cucumber, similar necrotic lesions and specific expression of Cssgr, as along with defense response-related genes (CsPR1 and CsLecRK6.1), were observed. This study suggests that the interaction between Cssgr and CSEP30^∆SP could trigger cell death and defense response, offering new insights into the molecular function of Cssgr in disease resistance in Gy14 cucumber.

Supplementary information: The online version contains supplementary material available at 10.1007/s11032-024-01504-6.

Keywords: Candidate secreted effector proteins; Cucumis sativus; Podosphaera xanthii; Powdery mildew; STAY-GREEN.