Expression Profiling of Botrytis cinerea Genes Identifies Three Patterns of Up-regulation in Planta and an FKBP12 Protein Affecting Pathogenicity

Gioti, Αnastasia; Simon, Adeline; Pêcheur, Pascal Le; Giraud, C.; Pradier, Jean‐Marc; Viaud, Muriel; Levis, Caroline

doi:10.1016/j.jmb.2006.01.076

Cited by 44 publications

(46 citation statements)

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“…The gene BcPIE3 was a good candidate for a B. cinerea virulence determinant, due to its expression profile in planta, revealed by previous results from both macroarray and qRT-PCR experiments (18) and by in vitro appressorium formation in qRT-PCR experiments in this study. The virulence of the ⌬36 mutant was severely reduced compared to that of the wild-type strain T4 (Fig.…”

Section: Bcpie3 Is a Virulence Factor Required For Lesion Expansionsupporting

confidence: 52%

“…In a previous study, the BcPIE3 gene was shown to be up-regulated in the early stage of Arabidopsis thaliana leaf infections with the B. cinerea T4 strain (18). Here, we undertook qRT-PCR analysis of cDNAs synthesized from RNA extracted from appressoria of strain T4.…”

Section: Resultsmentioning

confidence: 99%

“…Conditions tested (columns) were as follows: stage of appressorium formation on Teflon filters (see Materials and Methods), three consecutive stages of Botrytis in planta growth on Arabidopsis leaves (early stage, outset of colonization, and complete-colonization stage), and one stage of in vitro growth on a simple medium. These stages, the RNA preparation method, and identities of constitutive genes have been detailed by Gioti et al (18). Apart from BcPIE3, additional genes (rows) presented here as controls code for a putative methionine synthase (meth_synthase; AL14398), down-regulated in planta; a fungal tetraspanin named BcPLS1, specifically expressed in appressoria; and an FKBP12 protein, up-regulated in planta at the complete-colonization stage (18).…”

Section: Resultsmentioning

confidence: 99%

“…It is noteworthy that the BcPIE3 gene shows maximal expression at the early stage of infection, that is, 24 to 29 h postinoculation (18), whereas the mutant's phenotype is apparent during the expansion of necrotic lesions. This stage, previously defined as the outset of colonization, occurs at 33 to 48 h postinoculation (18).…”

Section: Bcpie3 Is a Virulence Factor Required For Lesion Expansionmentioning

confidence: 99%

“…In order to identify novel pathogenicity determinants, we recently studied the expression profiles of B. cinerea genes during the infection of Arabidopsis thaliana leaves. Among the genes shown to be induced in planta, one called B. cinerea PIE3 (BcPIE3) was found to be highly expressed 24 to 29 h after the inoculation of the fungus onto the plant leaves (18). To gain insight into early infection events, this gene was further studied.…”

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ABotrytis cinereaEmopamil Binding Domain Protein, Required for Full Virulence, Belongs to a Eukaryotic Superfamily Which Has Expanded in Euascomycetes

et al. 2008

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A previous transcriptomic analysis of 3,032 fungal genes identified the Botrytis cinerea PIE3 (BcPIE3) gene to be up-regulated early in planta (A. Gioti, A. Simon, P. Le Pêcheur, C. Giraud, J. M. Pradier, M. Viaud, and C. Levis, J. Mol. Biol. 358:372-386, 2006). In the present study, BcPIE3 was disrupted in order to determine its implication in pathogenicity. BcPIE3 was shown to be a virulence factor, since the ⌬BcPIE3 mutant was blocked during the colonization of tomato and bean leaves, giving lesions reduced in size by at least 74%. Within the emopamil binding domain (EBD), BcPIE3 shows significant structural similarities to mammalian emopamil binding proteins (EBPs). Mammalian EBPs function as sterol isomerases, but an analysis of the sterol content and the results of growth inhibition experiments with the ⌬BcPIE3 strain indicated that BcPIE3 is dispensable for ergosterol biosynthesis. The systematic identification of EBD-containing proteins included in public databases showed that these proteins constitute a protein superfamily present only in eukaryotes. Phylogenetic analysis showed that the ancestral EBD-encoding gene was duplicated in the common ancestor of animals and fungi after the split from plants. Finally, we present evidence that the EBP phylogenetic clade of this superfamily has further expanded exclusively in euascomycetes, especially in B. cinerea, which contains three copies of the EBP gene.Botrytis cinerea is a ubiquitous fungus capable of infecting a wide range of plants, fruits, and ornamentals at any stage of their development (15,30). In order to identify novel pathogenicity determinants, we recently studied the expression profiles of B. cinerea genes during the infection of Arabidopsis thaliana leaves. Among the genes shown to be induced in planta, one called B. cinerea PIE3 (BcPIE3) was found to be highly expressed 24 to 29 h after the inoculation of the fungus onto the plant leaves (18). To gain insight into early infection events, this gene was further studied.BcPIE3 showed strong similarity to mammalian emopamil binding proteins (EBPs). EBPs are integral, nonglycosylated proteins of the endoplasmic reticulum membrane, characterized by the presence of four transmembrane (TM) segments forming a binding domain for the drug emopamil, a phenylalkylamine Ca 2ϩ antagonist. This domain is called the emopamil binding domain (EBD). Mammalian EBPs (43, 25) have received increasing attention due to their binding properties. Indeed, the binding of sigma ligands but also of numerous, structurally diverse drugs and fungicides on the EBD (36) exerts anti-ischemic effects in animal models (43). Mammalian EBPs have been further studied because of a number of syndromes caused by EBP gene mutations, such as the genetic disorder X-linked dominant chondrodysplasia punctata, Conradi-Hünermann syndrome (5, 26), CHILD (congenital hemidysplasia with ichthyosiform erythroderma and limb defects) syndrome (21), and the "tattered" mouse phenotype (13).According to the above-mentioned studies, the molecular patholo...

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Section: Bcpie3 Is a Virulence Factor Required For Lesion Expansionsupporting

confidence: 52%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Bcpie3 Is a Virulence Factor Required For Lesion Expansionmentioning

confidence: 99%

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confidence: 99%

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