scholarly journals Analysis of the Molecular Dialogue Between Gray Mold (Botrytis cinerea) and Grapevine (Vitis vinifera) Reveals a Clear Shift in Defense Mechanisms During Berry Ripening

2015 ◽  
Vol 28 (11) ◽  
pp. 1167-1180 ◽  
Author(s):  
Jani Kelloniemi ◽  
Sophie Trouvelot ◽  
Marie-Claire Héloir ◽  
Adeline Simon ◽  
Bérengère Dalmais ◽  
...  
Keyword(s):  
Cell Wall ◽  
Botrytis Cinerea ◽  
Defense Mechanisms ◽  
Plant Cell Wall ◽  
Quantitative Polymerase Chain Reaction ◽  
Gray Mold ◽  
Ros Generation ◽  
Genome Wide ◽  
Mold Fungus ◽  
Berry Ripening

Mature grapevine berries at the harvesting stage (MB) are very susceptible to the gray mold fungus Botrytis cinerea, while veraison berries (VB) are not. We conducted simultaneous microscopic and transcriptomic analyses of the pathogen and the host to investigate the infection process developed by B. cinerea on MB versus VB, and the plant defense mechanisms deployed to stop the fungus spreading. On the pathogen side, our genome-wide transcriptomic data revealed that B. cinerea genes upregulated during infection of MB are enriched in functional categories related to necrotrophy, such as degradation of the plant cell wall, proteolysis, membrane transport, reactive oxygen species (ROS) generation, and detoxification. Quantitative-polymerase chain reaction on a set of representative genes related to virulence and microscopic observations further demonstrated that the infection is also initiated on VB but is stopped at the penetration stage. On the plant side, genome-wide transcriptomic analysis and metabolic data revealed a defense pathway switch during berry ripening. In response to B. cinerea inoculation, VB activated a burst of ROS, the salicylate-dependent defense pathway, the synthesis of the resveratrol phytoalexin, and cell-wall strengthening. On the contrary, in infected MB, the jasmonate-dependent pathway was activated, which did not stop the fungal necrotrophic process.

scholarly journals The VELVET Complex in the Gray Mold Fungus Botrytis cinerea: Impact of BcLAE1 on Differentiation, Secondary Metabolism, and Virulence

2015 ◽  
Vol 28 (6) ◽  
pp. 659-674 ◽  
Author(s):  
Julia Schumacher ◽  
Adeline Simon ◽  
Kim C. Cohrs ◽  
Stefanie Traeger ◽  
Antoine Porquier ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Natural Variation ◽  
Target Genes ◽  
Gray Mold ◽  
Carbohydrate Active Enzymes ◽  
Genome Wide ◽  
Mold Fungus ◽  
Velvet Complex ◽  
Genome Wide Expression ◽  
Insight Into

Botrytis cinerea, the gray mold fungus, is an important plant pathogen. Field populations are characterized by variability with regard to morphology, the mode of reproduction (conidiation or sclerotia formation), the spectrum of secondary metabolites (SM), and virulence. Natural variation in bcvel1 encoding the ortholog of Aspergillus nidulans VeA, a member of the VELVET complex, was previously shown to affect light-dependent differentiation, the formation of oxalic acid (OA), and virulence. To gain broader insight into the B. cinerea VELVET complex, an ortholog of A. nidulans LaeA, BcLAE1, a putative interaction partner of BcVEL1, was studied. BcVEL1 but not its truncated versions interacts with BcLAE1 and BcVEL2 (VelB ortholog). In accordance with the expected common as well as specific functions of BcVEL1 and BcLAE1, the deletions of both genes result in similar though not identical phenotypes. Both mutants lost the ability to produce OA, to colonize the host tissue, and to form sclerotia. However, mutants differ with regard to aerial hyphae and conidia formation. Genome-wide expression analyses revealed that BcVEL1 and BcLAE1 have common and distinct target genes. Some of the genes that are underexpressed in both mutants, e.g., those encoding SM-related enzymes, proteases, and carbohydrate-active enzymes, may account for their reduced virulence.

scholarly journals Cyclophilin BcCyp2 Regulates Infection-Related Development to Facilitate Virulence of the Gray Mold Fungus Botrytis cinerea

2021 ◽  
Vol 22 (4) ◽  
pp. 1694
Author(s):  
Jiao Sun ◽  
Chen-Hao Sun ◽  
Hao-Wu Chang ◽  
Song Yang ◽  
Yue Liu ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Gray Mold ◽  
Gene Encoding ◽  
Histone 3 ◽  
Mold Fungus ◽  
Related Development ◽  
Hyphal Morphology ◽  
Cellular Components

Cyclophilin (Cyp) and Ca2+/calcineurin proteins are cellular components related to fungal morphogenesis and virulence; however, their roles in mediating the pathogenesis of Botrytis cinerea, the causative agent of gray mold on over 1000 plant species, remain largely unexplored. Here, we show that disruption of cyclophilin gene BcCYP2 did not impair the pathogen mycelial growth, osmotic and oxidative stress adaptation as well as cell wall integrity, but delayed conidial germination and germling development, altered conidial and sclerotial morphology, reduced infection cushion (IC) formation, sclerotial production and virulence. Exogenous cyclic adenosine monophosphate (cAMP) rescued the deficiency of IC formation of the ∆Bccyp2 mutants, and exogenous cyclosporine A (CsA), an inhibitor targeting cyclophilins, altered hyphal morphology and prevented host-cell penetration in the BcCYP2 harboring strains. Moreover, calcineurin-dependent (CND) genes are differentially expressed in strains losing BcCYP2 in the presence of CsA, suggesting that BcCyp2 functions in the upstream of cAMP- and Ca2+/calcineurin-dependent signaling pathways. Interestingly, during IC formation, expression of BcCYP2 is downregulated in a mutant losing BcJAR1, a gene encoding histone 3 lysine 4 (H3K4) demethylase that regulates fungal development and pathogenesis, in B. cinerea, implying that BcCyp2 functions under the control of BcJar1. Collectively, our findings provide new insights into cyclophilins mediating the pathogenesis of B. cinerea and potential targets for drug intervention for fungal diseases.

scholarly journals The Xylanase Inhibitor TAXI-I Increases Plant Resistance to Botrytis cinerea by Inhibiting the BcXyn11a Xylanase Necrotizing Activity

Plants ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 601
Author(s):  
Silvio Tundo ◽  
Maria Chiara Paccanaro ◽  
Ibrahim Elmaghraby ◽  
Ilaria Moscetti ◽  
Renato D’Ovidio ◽  
...  
Keyword(s):  
Cell Wall ◽  
Botrytis Cinerea ◽  
Plant Cell Wall ◽  
Wild Type ◽  
Cell Wall Degrading Enzymes ◽  
Targeted Deletion ◽  
Plant Infection ◽  
Transgenic Lines ◽  
Xylanase Inhibitor ◽  
Main Component

During host plant infection, pathogens produce a wide array of cell wall degrading enzymes (CWDEs) to break the plant cell wall. Among CWDEs, xylanases are key enzymes in the degradation of xylan, the main component of hemicellulose. Targeted deletion experiments support the direct involvement of the xylanase BcXyn11a in the pathogenesis of Botrytis cinerea. Since the Triticum aestivum xylanase inhibitor-I (TAXI-I) has been shown to inhibit BcXyn11a, we verified if TAXI-I could be exploited to counteract B. cinerea infections. With this aim, we first produced Nicotiana tabacum plants transiently expressing TAXI-I, observing increased resistance to B. cinerea. Subsequently, we transformed Arabidopsis thaliana to express TAXI-I constitutively, and we obtained three transgenic lines exhibiting a variable amount of TAXI-I. The line with the higher level of TAXI-I showed increased resistance to B. cinerea and the absence of necrotic lesions when infiltrated with BcXyn11a. Finally, in a droplet application experiment on wild-type Arabidopsis leaves, TAXI-I prevented the necrotizing activity of BcXyn11a. These results would confirm that the contribution of BcXyn11a to virulence is due to its necrotizing rather than enzymatic activity. In conclusion, our experiments highlight the ability of the TAXI-I xylanase inhibitor to counteract B. cinerea infection presumably by preventing the necrotizing activity of BcXyn11a.

scholarly journals The BMP1 Gene Is Essential for Pathogenicity in the Gray Mold Fungus Botrytis cinerea

2000 ◽  
Vol 13 (7) ◽  
pp. 724-732 ◽  
Author(s):  
Li Zheng ◽  
Mathew Campbell ◽  
Jennifer Murphy ◽  
Stephen Lam ◽  
Jin-Rong Xu
Keyword(s):  
Growth Rate ◽  
Map Kinase ◽  
Botrytis Cinerea ◽  
Magnaporthe Grisea ◽  
Pathogenic Fungi ◽  
Gray Mold ◽  
Necrotrophic Pathogen ◽  
Kinase Gene ◽  
Plant Infection ◽  
Mold Fungus

In Magnaporthe grisea, a well-conserved mitogen-activated protein (MAP) kinase gene, PMK1, is essential for fungal pathogenesis. In this study, we tested whether the same MAP kinase is essential for plant infection in the gray mold fungus Botrytis cinerea, a necrotrophic pathogen that employs infection mechanisms different from those of M. grisea. We used a polymerase chain reaction-based approach to isolate MAP kinase homologues from B. cinerea. The Botrytis MAP kinase required for pathogenesis (BMP) MAP kinase gene is highly homologous to the M. grisea PMK1. BMP1 is a single-copy gene. bmp1 gene replacement mutants produced normal conidia and mycelia but were reduced in growth rate on nutrient-rich medium. bmp1 mutants were nonpathogenic on carnation flowers and tomato leaves. Re-introduction of the wild-type BMP1 allele into the bmp1 mutant restored both normal growth rate and pathogenicity. Further studies indicated that conidia from bmp1 mutants germinated on plant surfaces but failed to penetrate and macerate plant tissues. bmp1 mutants also appeared to be defective in infecting through wounds. These results indicated that BMP1 is essential for plant infection in B. cinerea, and this MAP kinase pathway may be widely conserved in pathogenic fungi for regulating infection processes.

scholarly journals Identification of Pathogenesis-Associated Genes by T-DNA–Mediated Insertional Mutagenesis in Botrytis cinerea: A Type 2A Phosphoprotein Phosphatase and an SPT3 Transcription Factor Have Significant Impact on Virulence

2012 ◽  
Vol 25 (4) ◽  
pp. 481-495 ◽  
Author(s):  
S. Giesbert ◽  
J. Schumacher ◽  
V. Kupas ◽  
J. Espino ◽  
N. Segmüller ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Protein Phosphatase ◽  
Insertional Mutagenesis ◽  
Single Copy ◽  
Gene Replacement ◽  
Gray Mold ◽  
Dna Integration ◽  
Bean Plants ◽  
Mold Fungus ◽  
Border Sequence

Agrobacterium tumefaciens–mediated transformation (ATMT) was used to generate an insertional mutant library of the gray mold fungus Botrytis cinerea. From a total of 2,367 transformants, 68 mutants showing significant reduction in virulence on tomato and bean plants were analyzed in detail. As reported for other fungal ATMT libraries, integrations were mostly single copy, occurred preferentially in noncoding (regulatory) regions, and were frequently accompanied by small deletions of the target sequences and loss of parts of the border sequence. Two T-DNA integration events that were found to be linked to virulence were characterized in more detail: a catalytic subunit of a PP2A serine/threonine protein phosphatase (BcPP2Ac) and the SPT3 subunit of a Spt-Ada-Gcn5-acetyltransferase (SAGA-like) transcriptional regulator complex. Gene replacement and silencing approaches revealed that both Bcpp2Ac and SPT3 are crucial for virulence, growth, and differentiation as well as for resistance to H2O2 in B. cinerea.

scholarly journals Involvement of the Autophagy Protein Atg6 in Development and Virulence in the Gray Mold Fungus Botrytis cinerea

2021 ◽  
Vol 12 ◽  
Author(s):  
Na Liu ◽  
Shanyue Zhou ◽  
Baohua Li ◽  
Weichao Ren
Keyword(s):  
Botrytis Cinerea ◽  
Biological Function ◽  
Gray Mold ◽  
Economic Losses ◽  
Related Protein ◽  
Targeted Deletion ◽  
Cellular Processes ◽  
Mold Fungus ◽  
Evolutionarily Conserved ◽  
Sclerotial Formation

Gray mold caused by Botrytis cinerea is a devastating disease that leads to huge economic losses worldwide. Autophagy is an evolutionarily conserved process that maintains intracellular homeostasis through self-eating. In this study, we identified and characterized the biological function of the autophagy-related protein Atg6 in B. cinerea. Targeted deletion of the BcATG6 gene showed block of autophagy and several phenotypic defects in aspects of mycelial growth, conidiation, sclerotial formation and virulence. All of the phenotypic defects were restored by targeted gene complementation. Taken together, these results suggest that BcAtg6 plays important roles in the regulation of various cellular processes in B. cinerea.

scholarly journals Genome-wide and enzymatic analysis reveals efficient D-galacturonic acid metabolism in the basidiomycete yeastRhodosporidium toruloides

2019 ◽  
Author(s):  
Ryan J. Protzko ◽  
Christina A. Hach ◽  
Samuel T. Coradetti ◽  
Magdalena A. Hackhofer ◽  
Sonja Magosch ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Galacturonic Acid ◽  
Plant Cell Wall ◽  
Rhodosporidium Toruloides ◽  
Cell Wall Polysaccharide ◽  
Aromatic Molecules ◽  
Renewable Feedstocks ◽  
Genome Wide ◽  
A Genome

AbstractBiorefining of renewable feedstocks is one of the most promising routes to replace fossil-based products. Since many common fermentation hosts, such asSaccharomyces cerevisiae, are naturally unable to convert many component plant cell wall polysaccharides, the identification of organisms with broad catabolism capabilities represents an opportunity to expand the range of substrates used in fermentation biorefinery approaches. The red basidiomycete yeastRhodosporidium toruloidesis a promising and robust host for lipid and terpene derived chemicals. Previous studies demonstrated assimilation of a range of substrates, from C5/C6-sugars to aromatic molecules similar to lignin monomers. In the current study, we analyzedR. toruloidespotential to assimilate D-galacturonic acid, a major sugar in many pectin-rich agricultural waste streams, including sugar beet pulp and citrus peels. D-galacturonic acid is not a preferred substrate for many fungi, but its metabolism was found to be on par with D-glucose and D-xylose inR. toruloides. A genome-wide analysis by combined RNAseq/RB-TDNAseq revealed those genes with high relevance for fitness on D-galacturonic acid. WhileR. toruloideswas found to utilize the same non-phosphorylative catabolic pathway known from ascomycetes, the maximal velocities of several enzymes exceeded those previously reported. In addition, an efficient downstream glycerol catabolism and a novel transcription factor were found to be important for D-galacturonic acid utilization. These results set the basis for use ofR. toruloidesas a potential host for pectin-rich waste conversions and demonstrate its suitability as a model for metabolic studies in basidiomycetes.ImportanceThe switch from the traditional fossil-based industry to a green and sustainable bio-economy demands the complete utilization of renewable feedstocks. Many currently used bio-conversion hosts are unable to utilize major components of plant biomass, warranting the identification of microorganisms with broader catabolic capacity and characterization of their unique biochemical pathways. D-galacturonic acid is a plant component of bio-conversion interest and is the major backbone sugar of pectin, a plant cell wall polysaccharide abundant in soft and young plant tissues. The red basidiomycete and oleaginous yeastRhodosporidium toruloideshas been previously shown to utilize a range of sugars and aromatic molecules. Using state-of-the-art functional genomic methods, physiological and biochemical assays, we elucidated the molecular basis underlying the efficient metabolism of D-galacturonic acid. This study identifies an efficient pathway for uronic acid conversion to guide future engineering efforts, and represents the first detailed metabolic analysis of pectin metabolism in a basidiomycete fungus.

scholarly journals Expression of Putative Defense Responses in Cannabis Primed by Pseudomonas and/or Bacillus Strains and Infected by Botrytis cinerea

2020 ◽  
Vol 11 ◽  
Author(s):  
Carole Balthazar ◽  
Gabrielle Cantin ◽  
Amy Novinscak ◽  
David L. Joly ◽  
Martin Filion
Keyword(s):  
Botrytis Cinerea ◽  
Cannabis Sativa ◽  
Molecular Level ◽  
Quantitative Polymerase Chain Reaction ◽  
Defense Responses ◽  
Gray Mold ◽  
Control Measures ◽  
Effective Control ◽  
Defense Genes ◽  
Polymerase Chain

Cannabis (Cannabis sativa L.) offers many industrial, agricultural, and medicinal applications, but is commonly threatened by the gray mold disease caused by the fungus Botrytis cinerea. With few effective control measures currently available, the use of beneficial rhizobacteria represents a promising biocontrol avenue for cannabis. To counter disease development, plants rely on a complex network of inducible defense pathways, allowing them to respond locally and systemically to pathogens attacks. In this study, we present the first attempt to control gray mold in cannabis using beneficial rhizobacteria, and the first investigation of cannabis defense responses at the molecular level. Four promising Pseudomonas (LBUM223 and WCS417r) and Bacillus strains (LBUM279 and LBUM979) were applied as single or combined root treatments to cannabis seedlings, which were subsequently infected by B. cinerea. Symptoms were recorded and the expression of eight putative defense genes was monitored in leaves by reverse transcription quantitative polymerase chain reaction. The rhizobacteria did not significantly control gray mold and all infected leaves were necrotic after a week, regardless of the treatment. Similarly, no systemic activation of putative cannabis defense genes was reported, neither triggered by the pathogen nor by the rhizobacteria. However, this work identified five putative defense genes (ERF1, HEL, PAL, PR1, and PR2) that were strongly and sustainably induced locally at B. cinerea’s infection sites, as well as two stably expressed reference genes (TIP41 and APT1) in cannabis. These markers will be useful in future researches exploring cannabis defense pathways.

Identification of SSR markers associated with saccharification yield using pool-based genome-wide association mapping in sorghum

Genome ◽  
2011 ◽  
Vol 54 (11) ◽  
pp. 883-889 ◽  
Author(s):  
Yi-Hong Wang ◽  
Durga D. Poudel ◽  
Karl H. Hasenstein
Keyword(s):  
Cell Wall ◽  
Association Mapping ◽  
Ssr Markers ◽  
Plant Cell Wall ◽  
Linear Models ◽  
Plant Biomass ◽  
Genome Wide Association ◽  
Whole Genome Sequence ◽  
Genome Wide ◽  
Saccharification Yield

Saccharification describes the conversion of plant biomass by cellulase into glucose. Because plants have never been selected for high saccharification yield, cellulosic ethanol production faces a significant bottleneck. To improve saccharification yield, it is critical to identify the genes that affect this process. In this study, we used pool-based genome-wide association mapping to identify simple sequence repeat (SSR) markers associated with saccharification yield. Screening of 703 SSR markers against the low and high saccharification pools identified two markers on the sorghum chromosomes 2 (23-1062) and 4 (74-508c) associated with saccharification yield. The association was significant at 1% using either general or mixed linear models. Localization of these markers based on the whole genome sequence indicates that 23-1062 is 223 kb from a β–glucanase (Bg) gene and 74-508c is 81 kb from a steroid-binding protein (Sbp) gene. Bg is critical for cell wall assembly and degradation, but Sbp can suppress the expression of Bg as demonstrated in Arabidopsis (Yang et al. 2005). These markers are found physically close to genes encoding plant cell wall synthesis enzymes such as xyloglucan fucosyltransferase (149 kb from 74-508c) and UDP-d-glucose 4-epimerase (46 kb from 23-1062). Genetic transformation of selected candidate genes is in progress to examine their effect on saccharification yield in plants.

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