20.500.12556/RUL-128210 Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion The vascular plant pathogen Verticillium nonalfalfae causes Verticillium wilt in several important crops. VnaSSP4.2 was recently discovered as a V. nonalfalfae virulence effector protein in the xylem sap of infected hop. Here, we expanded our search for candidate secreted effector proteins (CSEPs) in the V. nonalfalfae predicted secretome using a bioinformatic pipeline built on V. nonalfalfae genome data, RNA-Seq and proteomic studies of the interaction with hop. The secretome, rich in carbohydrate active enzymes, proteases, redox proteins and proteins involved in secondary metabolism, cellular processing and signaling, includes 263 CSEPs. Several homologs of known fungal effectors (LysM, NLPs, Hce2, Cerato-platanins, Cyanovirin-N lectins, hydrophobins and CFEM domain containing proteins) and avirulence determinants in the PHI database (Avr-Pita1 and MgSM1) were found. The majority of CSEPs were non-annotated and were narrowed down to 44 top priority candidates based on their likelihood of being effectors. These were examined by spatio-temporal gene expression profiling of infected hop. Among the highest in planta expressed CSEPs, five deletion mutants were tested in pathogenicity assays. A deletion mutant of VnaUn.279, a lethal pathotype specific gene with sequence similarity to SAM-dependent methyltransferase (LaeA), had lower infectivity and showed highly reduced virulence, but no changes in morphology, fungal growth or conidiation were observed. Several putative secreted effector proteins that probably contribute to V. nonalfalfae colonization of hop were identified in this study. Among them, LaeA gene homolog was found to act as a potential novel virulence effector of V. nonalfalfae. The combined results will serve for future characterization of V. nonalfalfae effectors, which will advance our understanding of Verticillium wilt disease. plant fungal pathogens protein domains Verticillium fungal genomics fungal genetics proteases plant pathogens gene expression Verticillium nonalfalfae hmelj genetika true false true Angleški jezik Slovenski jezik Članek v reviji 2021-07-06 15:00:35 2021-07-06 15:00:36 2022-09-07 04:13:42 0000-00-00 00:00:00 2018 0 0 28 str. iss. 6, art. e0198971 Vol. 13 2018 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 1932-6203 10.1371/journal.pone.0198971 8940665 RAZ_Marton_Kristina_2018.pdf RAZ_Marton_Kristina_2018.pdf 1 C02D1291A059D4BE28B2264DEC7C494C 1799556811d7f60bf3b117ff8c30ba6bd252550dbe602b59a3ba3cc97df25535 a09a1497-de5d-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=144876 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0198971 0 95a43d2b-de5d-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=144875 Biotehniška fakulteta 0 0 0