A putative phospholipase C is involved in Pichia fermentans dimorphic transition

• Published in: Biochimica et biophysica acta Vol. 1840; no. 1; pp. 344 - 349
• Main Authors: Sanna, Maria Lina, Zara, Giacomo, Zara, Severino, Migheli, Quirico, Budroni, Marilena, Mannazzu, Ilaria
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2014
• Subjects: 3′-5′-cyclic adenosine monophosphate (cAMP); antagonists; biological control; Cloning, Molecular; complementary DNA; cyclic AMP; Dimorphic shift; DNA, Fungal - genetics; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Gene Library; gene overexpression; genes; Hyphae - enzymology; Hyphae - genetics; Hyphae - growth & development; Hyphae - pathogenicity; Image Processing, Computer-Assisted; methionine; Monilinia; nucleotide sequences; pathogenicity; Phospholipase C; Pichia - enzymology; Pichia - genetics; Pichia - growth & development; Pichia - pathogenicity; Pichia fermentans; plant pathogens; Pseudohypha; Rapid subtraction hybridization (RaSH); Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; risk; RNA; RNA, Messenger - genetics; signal transduction; Subtraction Technique; suppression subtractive hybridization; Type C Phospholipases - genetics; Type C Phospholipases - metabolism; yeasts; Pseudohypha; Rapid subtraction hybridization (RaSH); Pichia fermentans; Dimorphic shift; 3′-5′-cyclic adenosine monophosphate (cAMP); Phospholipase C
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2013.09.030

Pichia fermentans DiSAABA 726 is a dimorphic yeast that reversibly shifts from yeast-like to pseudohyphal morphology. This yeast behaves as a promising antagonist of Monilia spp. in the yeast-like form, but becomes a destructive plant pathogen in the pseudohyphal form thus raising the problem of the biological risk associated with the use of dimorphic yeasts as microbial antagonists in the biocontrol of phytopathogenic fungi. Pichia fermentans DiSAABA 726 was grown in urea- and methionine-containing media in order to induce and separate yeast-like and pseudohyphal morphologies. Total RNA was extracted from yeast-like cells and pseudohyphae and retro-transcribed into cDNA. A rapid subtraction hybridization approach was utilized to obtain the cDNA sequences putatively over-expressed during growth on methionine-containing medium and involved in pseudohyphal transition. Five genes that are over-expressed during yeast-like/pseudohyphal dimorphic transition were isolated. One of these, encoding a putative phospholipase C, is involved in P. fermentans filamentation. In fact, while the inhibition of phospholipase C, by means of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphorylcholine (Et-18), is accompanied by a significant reduction of pseudohyphae formation in P. fermentans, the addition of exogenous cAMP fully restores pseudohyphal growth also in the presence of Et-18. Phospholipase C is part of a putative “methionine sensing machinery” that activates cAMP-PKA signal transduction pathway and controls P. fermentans yeast-like/pseudohyphal dimorphic transition. Phospholipase C is a promising molecular target for further investigations into the link between pseudohyphae formation and pathogenicity in P. fermentans. •DiSAABA 726 reversibly shifts from yeast-like to pseudohyphal morphologies.•Pathogenic behavior of DiSAABA 726 is accompanied by pseudohyphal morphology.•Inhibition of phospholipase C affects pseudohyphal transition.•Exogenous cAMP availability circumvents phospholipase C impairment.