Uriel Maor - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel; Institute of Biochemistry, Food Science and Nutrition, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel.
Omer Barda - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Sudharsan Sadhasivam - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Yang Bi - College of Food Science and Engineering, Gansu Agricultural University, Lanzhou, China.
Elena Levin - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Varda Zakin - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Dov B Prusky - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israe; College of Food Science and Engineering, Gansu Agricultural University, Lanzhou, China.
Edward Sionov - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Aspergillus carbonarius is the major producer of ochratoxin A (OTA) among Aspergillus species, but the contribution of this secondary metabolite to fungal virulence has not been assessed. We characterized the functions and addressed the roles of three factors in the regulation of OTA synthesis and pathogenicity in A. carbonarius: LaeA, a transcriptional factor regulating the production of secondary metabolites; polyketide synthase, required for OTA biosynthesis; and glucose oxidase (GOX), regulating gluconic acid (GLA) accumulation and acidification of the host tissue during fungal growth. Deletion of laeA in A. carbonarius resulted in significantly reduced OTA production in colonized nectarines and grapes. The ∆laeA mutant was unable to efficiently acidify the colonized tissue, as a direct result of diminished GLA production, leading to attenuated virulence in infected fruit compared to the wild type (WT). The designed Acpks-knockout mutant resulted in complete inhibition of OTA production in vitro and in colonized fruit. Interestingly, physiological analysis revealed that the colonization pattern of the ∆Acpks mutant was similar to that of the WT strain, with high production of GLA in the colonized tissue, suggesting that OTA accumulation does not contribute to A. carbonarius pathogenicity. Disruption of the Acgox gene inactivated GLA production in A. carbonarius, and this mutant showed attenuated virulence in infected fruit compared to the WT strain. These data identify the global regulator LaeA and GOX as critical factors modulating A. carbonarius pathogenicity by controlling transcription of genes important for fungal secondary metabolism and infection.
Uriel Maor - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel; Institute of Biochemistry, Food Science and Nutrition, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel.
Omer Barda - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Sudharsan Sadhasivam - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Yang Bi - College of Food Science and Engineering, Gansu Agricultural University, Lanzhou, China.
Elena Levin - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Varda Zakin - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Dov B Prusky - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israe; College of Food Science and Engineering, Gansu Agricultural University, Lanzhou, China.
Edward Sionov - Institute of Postharvest and Food Sciences, The Volcani Center, Agricultural Research Organization, Rishon LeZion, Israel.
Aspergillus carbonarius is the major producer of ochratoxin A (OTA) among Aspergillus species, but the contribution of this secondary metabolite to fungal virulence has not been assessed. We characterized the functions and addressed the roles of three factors in the regulation of OTA synthesis and pathogenicity in A. carbonarius: LaeA, a transcriptional factor regulating the production of secondary metabolites; polyketide synthase, required for OTA biosynthesis; and glucose oxidase (GOX), regulating gluconic acid (GLA) accumulation and acidification of the host tissue during fungal growth. Deletion of laeA in A. carbonarius resulted in significantly reduced OTA production in colonized nectarines and grapes. The ∆laeA mutant was unable to efficiently acidify the colonized tissue, as a direct result of diminished GLA production, leading to attenuated virulence in infected fruit compared to the wild type (WT). The designed Acpks-knockout mutant resulted in complete inhibition of OTA production in vitro and in colonized fruit. Interestingly, physiological analysis revealed that the colonization pattern of the ∆Acpks mutant was similar to that of the WT strain, with high production of GLA in the colonized tissue, suggesting that OTA accumulation does not contribute to A. carbonarius pathogenicity. Disruption of the Acgox gene inactivated GLA production in A. carbonarius, and this mutant showed attenuated virulence in infected fruit compared to the WT strain. These data identify the global regulator LaeA and GOX as critical factors modulating A. carbonarius pathogenicity by controlling transcription of genes important for fungal secondary metabolism and infection.