Calcineurin phosphatase and phospholipase C are required for developmental and pathological functions in the citrus fungal pathogen Alternaria alternata.
Excessive Ca(2+) or compounds interfering with phosphoinositide cycling have been found to inhibit the growth of the tangerine pathotype of Alternaria alternata, suggesting a crucial role of Ca(2+) homeostasis in this pathotype. The roles of PLC1, a phospholipase C-coding gene and CAL1, a calcineurin phosphatase-coding gene were investigated. Targeted gene disruption showed that both PLC1 and CAL1 were required for vegetative growth, conidial formation and pathogenesis in citrus. Fungal strains lacking PLC1 or CAL1 exhibited extremely slow growth and induced small lesions on calamondin leaves. Δplc1 mutants produced fewer conidia, which germinated at slower rates than wild-type. Δcal1 mutants produced abnormal hyphae and failed to produce any mature conidia, but instead produced highly melanized bulbous hyphae with distinct septae. Fluorescence microscopy using Fluo-3 dye as a Ca(2+) indicator revealed that the Δplc1 mutant hyphae emitted stronger cytosolic fluorescence, and the Δcal1 mutant hyphae emitted less cytosolic fluorescence, than those of wild-type. Infection assessed on detached calamondin leaves revealed that application of CaCl2 or neomycin 24 h prior to inoculation provided protection against Alt. alternata. These data indicate that a dynamic equilibrium of cellular Ca(2+) is critical for developmental and pathological processes of Alt. alternata.
Citrus Research and Education Center, Institute of Food and Agricultural Sciences (IFAS), University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.
Hsieh-Chin
HC
Tsai
Department of Plant Pathology, IFAS, University of Florida, Gainesville, FL 32611, USA.|Citrus Research and Education Center, Institute of Food and Agricultural Sciences (IFAS), University of Florida, 700 Experiment Station Road, Lake Alfred, FL 33850, USA.
Kuang-Ren
KR
Chung
D000528:Alternaria / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002957:Citrus / Q000382:microbiology*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D006706:Homeostasis; D006861:Hydrogen Peroxide / Q000378:metabolism; D025301:Hyphae; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics* / Q000378:metabolism; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000382:microbiology; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D013172:Spores, Fungal; D010738:Type C Phospholipases / Q000235:genetics* / Q000378:metabolism; D014774:Virulence
D005656:Fungal Proteins; D006861:Hydrogen Peroxide; C120817:calcineurin phosphatase; D010744:Phosphoric Monoester Hydrolases; D010738:Type C Phospholipases; D002118:Calcium