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“ Candidatus  Liberibacter asiaticus” (CLas), one of the causal agents of citrus Huanglongbing (HLB), secretes proteins with functions that are largely unknown. In this study, we demonstrated that CLIBASIA_00460, one of the CLas-encoded Sec-dependent presecretory proteins, might contribute to the phytopathogenicity of CLas.  CLIBASIA_00460  was conserved in CLas strains and expressed at a significantly higher level in citrus than in Asian citrus psyllid.  Agrobacteria -mediated transient expression in  Nicotiana benthamiana  epidermal cells showed that the mature CLIBASIA_00460 (m460) without the putative Sec-dependent signal peptide was localized in multiple cellular compartments including nucleus at 25°C, but that nuclear accumulation was greatly decreased as the temperature rose to 32°C. When overexpressed  via  a  Potato virus X  (PVX)-based expression vector in  N. benthamiana , m460 induced no local symptoms, but tiny necrotic spots were scattered on the systemic leaves. However, NLS-m460, which contains the SV40 nuclear localization sequence (NLS) at the N-terminus to promote nuclear import of m460, caused chlorosis and necrosis in the local leaves and severe necrosis in the systemic leaves. Taken together, these data suggest that CLIBASIA_00460 represented a novel virulence factor of CLas, and that nuclear localization of this protein was temperature dependent and positively correlated with its pathogenicity  in planta .

frontiers in microbiology

Nuclear Import of a Secreted “Candidatus Liberibacter asiaticus” Protein is Temperature Dependent and Contributes to Pathogenicity in Nicotiana benthamiana