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Int. J. Mol. Sci. 2017, 18(11), 2489; https://doi.org/10.3390/ijms18112489

Identification and Initial Characterization of the Effectors of an Anther Smut Fungus and Potential Host Target Proteins

1
Department of Biology, Program on Disease Evolution, University of Louisville, Louisville, KY 40208, USA
2
Department of Molecular Biology and University of South Florida Health Byrd Alzheimer’s Research Institute, Morsani College of Medicine, University of South Florida, Tampa, FL 33612, USA
3
Laboratory of New Methods in Biology, Institute for Biological Instrumentation, Russian Academy of Sciences, Institutskaya Str., 7, Pushchino, Moscow Region 142290, Russia
Current Address: Defence Medical and Environmental Research Institute, DSO National Laboratories, Singapore, Singapore.
*
Author to whom correspondence should be addressed.
Received: 24 October 2017 / Revised: 5 November 2017 / Accepted: 16 November 2017 / Published: 22 November 2017
(This article belongs to the Special Issue Plant Microbe Interaction 2017)
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Abstract

(1) Background: Plant pathogenic fungi often display high levels of host specificity and biotrophic fungi; in particular, they must manipulate their hosts to avoid detection and to complete their obligate pathogenic lifecycles. One important strategy of such fungi is the secretion of small proteins that serve as effectors in this process. Microbotryum violaceum is a species complex whose members infect members of the Caryophyllaceae; M. lychnidis-dioicae, a parasite on Silene latifolia, is one of the best studied interactions. We are interested in identifying and characterizing effectors of the fungus and possible corresponding host targets; (2) Methods: In silico analysis of the M. lychnidis-dioicae genome and transcriptomes allowed us to predict a pool of small secreted proteins (SSPs) with the hallmarks of effectors, including a lack of conserved protein family (PFAM) domains and also localized regions of disorder. Putative SSPs were tested for secretion using a yeast secretion trap method. We then used yeast two-hybrid analyses for candidate-secreted effectors to probe a cDNA library from a range of growth conditions of the fungus, including infected plants; (3) Results: Roughly 50 SSPs were identified by in silico analysis. Of these, 4 were studied further and shown to be secreted, as well as examined for potential host interactors. One of the putative effectors, MVLG_01732, was found to interact with Arabidopsis thaliana calcium-dependent lipid binding protein (AtCLB) and with cellulose synthase interactive protein 1 orthologues; and (4) Conclusions: The identification of a pool of putative effectors provides a resource for functional characterization of fungal proteins that mediate the delicate interaction between pathogen and host. The candidate targets of effectors, e.g., AtCLB, involved in pollen germination suggest tantalizing insights that could drive future studies. View Full-Text
Keywords: biotrophic pathogen; anther smut; fungal effectors; Microbotryum violaceum biotrophic pathogen; anther smut; fungal effectors; Microbotryum violaceum
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Kuppireddy, V.S.; Uversky, V.N.; Toh, S.S.; Tsai, M.-C.; Beckerson, W.C.; Cahill, C.; Carman, B.; Perlin, M.H. Identification and Initial Characterization of the Effectors of an Anther Smut Fungus and Potential Host Target Proteins. Int. J. Mol. Sci. 2017, 18, 2489.

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