scholarly journals Actin Is Required for Cellular Development and Virulence of Botrytis cinerea via the Mediation of Secretory Proteins

mSystems ◽  
2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Hua Li ◽  
Zhanquan Zhang ◽  
Guozheng Qin ◽  
Chang He ◽  
Boqiang Li ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Virulence Factors ◽  
Proteomic Analysis ◽  
Crucial Role ◽  
Pathogenic Fungi ◽  
Extracellular Proteins ◽  
Secretory Proteins ◽  
Wild Type ◽  
Cell Wall Degrading Enzymes ◽  
Vital Component

ABSTRACT Actin is a vital component of the cytoskeleton of living cells and is involved in several complex processes. However, its functions in plant-pathogenic fungi are largely unknown. In this paper, we found that deletion of the Botrytis cinerea actin gene bcactA reduced growth and sporulation of B. cinerea and lowered virulence. Based on iTRAQ (isobaric tags for relative and absolute quantification)-based proteomic analysis, we compared changes of the secretome in ΔbcactA and wild-type strains. A total of 40 proteins exhibited significant differences in abundance in ΔbcactA mutants compared with the wild type. These proteins included 11 down-accumulated cell wall-degrading enzymes (CWDEs). Among them, two CWDEs, cellobiohydrolase (BcCBH) and β-endoglucanase (BcEG), were found to contribute to the virulence of B. cinerea, indicating that bcactA plays a crucial role in regulating the secretion of extracellular virulence factors. These findings unveil previously unknown functions of BcactA to mediate the virulence of B. cinerea and provide new mechanistic insights into the role of BcactA in the complex pathogenesis of B. cinerea. IMPORTANCE The cytoskeleton is an important network that exists in cells of all domains of life. In eukaryotic cells, actin is a vital component of the cytoskeleton. Here, we report that BcactA, an actin protein in B. cinerea, can affect the growth, sporulation, and virulence of B. cinerea. Furthermore, iTRAQ-based proteomic analysis showed that BcactA affects the abundance of 40 extracellular proteins, including 11 down-accumulated CWDEs. Among them, two CWDEs, cellobiohydrolase (BcCBH) and β-endoglucanase (BcEG), contributed to the virulence of B. cinerea, indicating that bcactA plays a crucial role in regulating extracellular virulence factors. These findings unveil previously unknown functions of BcactA in mediating growth, sporulation, and virulence of B. cinerea.

scholarly journals Comparative Proteomics Reveals the Potential Targets of BcNoxR, a Putative Regulatory Subunit of NADPH Oxidase of Botrytis cinerea

2016 ◽  
Vol 29 (12) ◽  
pp. 990-1003 ◽  
Author(s):  
Hua Li ◽  
Zhanquan Zhang ◽  
Chang He ◽  
Guozheng Qin ◽  
Shiping Tian
Keyword(s):  
Nadph Oxidase ◽  
Botrytis Cinerea ◽  
Proteomic Analysis ◽  
Fungal Pathogens ◽  
Molecular Mechanisms ◽  
Quantitative Polymerase Chain Reaction ◽  
Tomato Fruit ◽  
Polymerase Chain Reaction Analysis ◽  
Regulatory Subunit ◽  
Wild Type

The NADPH oxidase (NOX) complex has been shown to play a crucial role in stress response and in the virulence of various fungal pathogens. The underlying molecular mechanisms of NOX, however, remain largely unknown. In the present study, a comparative proteomic analysis compared changes in protein abundance in wild-type Botrytis cinerea and ΔbcnoxR mutants in which the regulatory subunit of NOX was deleted. The ΔbcnoxR mutants exhibited reduced growth, sporulation, and impaired virulence. A total of 60 proteins, representing 49 individual genes, were identified in ΔbcnoxR mutants that exhibited significant differences in abundance relative to wild-type. Reverse transcription-quantitative polymerase chain reaction analysis demonstrated that the differences in transcript levels for 36 of the genes encoding the identified proteins were in agreement with the proteomic analysis, while the remainder exhibited reverse levels. Functional analysis of four proteins that decreased abundance in the ΔbcnoxR mutants indicated that 6-phosphogluconate dehydrogenase (BcPGD) played a role in the growth and sporulation of B. cinerea. The Δbcpgd mutants also displayed impaired virulence on various hosts, such as apple, strawberry, and tomato fruit. These results suggest that NOX can influence the expression of BcPGD, which has an impact on growth, sporulation, and virulence of B. cinerea.

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 Identification of Botrytis cinerea Genes Up-Regulated During Infection and Controlled by the Gα Subunit BCG1 Using Suppression Subtractive Hybridization (SSH)

2004 ◽  
Vol 17 (5) ◽  
pp. 537-546 ◽  
Author(s):  
Christian Schulze Gronover ◽  
Corinna Schorn ◽  
Bettina Tudzynski
Keyword(s):  
Botrytis Cinerea ◽  
Differentially Expressed Genes ◽  
Suppression Subtractive Hybridization ◽  
Target Genes ◽  
Subtractive Hybridization ◽  
Differentially Expressed ◽  
Wild Type ◽  
In Planta ◽  
Cell Wall Degrading Enzymes ◽  
Fungal Genes

The Gα subunit BCG1 plays an important role during the infection of host plants by Botrytis cinerea. Δbcg1 Mutants are able to conidiate, penetrate host leaves, and produce small primary lesions. However, in contrast to the wild type, the mutants completely stop invasion of plant tissue at this stage; secondary lesions have never been observed. Suppression subtractive hybridization (SSH) was used to identify fungal genes whose expression on the host plant is specifically affected in bcg1 mutants. Among the 22 differentially expressed genes, we found those which were predicted to encode proteases, enzymes involved in secondary metabolism, and others encoding cell wall-degrading enzymes. All these genes are highly expressed during infection in the wild type but not in the mutant. However, the genes are expressed in both the wild type and the mutant under certain conditions in vitro. Most of the BCG1-controlled genes are still expressed in adenylate cyclase (bac) mutants in planta, suggesting that BCG1 is involved in at least one additional signaling cascade in addition to the cAMP-depending pathway. In a second SSH approach, 1,500 clones were screened for those that are specifically induced by the wild type during the infection of bean leaves. Of the 22 BCG1-controlled genes, 11 also were found in the in planta SSH library. Therefore, SSH technology can be successfully applied to identify target genes of signaling pathways and differentially expressed genes in planta.

scholarly journals Knocking Out Bcsas1 in Botrytis cinerea Impacts Growth, Development, and Secretion of Extracellular Proteins, Which Decreases Virulence

2014 ◽  
Vol 27 (6) ◽  
pp. 590-600 ◽  
Author(s):  
Zhanquan Zhang ◽  
Guozheng Qin ◽  
Boqiang Li ◽  
Shiping Tian
Keyword(s):  
Botrytis Cinerea ◽  
Secretory Pathway ◽  
Pathogenic Fungi ◽  
Extracellular Proteins ◽  
Rab Gtpase ◽  
Rab Gtpases ◽  
Extracellular Medium ◽  
Expression Of Genes ◽  
Transport Vesicles ◽  
Genes Encoding

Pathogenic fungi usually secrete a series of virulence factors to the extracellular environment to facilitate infection. Rab GTPases play a central role in the secretory pathway. To explore the function of Rab/GTPase in filamentous fungi, we knocked out a Rab/GTPase family gene, Bcsas1, in Botrytis cinerea, an aggressive fungal pathogen that infects more than 200 plant species. A detailed analysis was conducted on the virulence and the secretory capability of the mutants. The results indicated that knockout of Bcsas1 inhibited hyphal development and reduced sporulation of B. cinerea on potato dextrose agar plates resulting in reduced virulence on various fruit hosts. Knocking out the Bcsas1 gene led to an accumulation of transport vesicles at the hyphal tip, significantly reduced extracellular protein content, and lowered the activity of polygalacturonase and xylanase in the extracellular medium. However, mutation of Bcsas1 did not affect the expression of genes encoding polygalacturonase and xylanase, suggesting the secretion of these two family enzymes was suppressed in the mutant. Moreover, a comparative analysis of the secretome provided further evidence that the disruption of Bcsas1 in mutant strains significantly depressed the secretion of polysaccharide hydrolases and proteases. The results indicate that Bcsas1, the Rab8/SEC4-like gene, plays a crucial role in development, protein secretion, and virulence of B. cinerea.

scholarly journals Comparative Proteomic Analysis of Extracellular Proteins of Edwardsiella tarda

2002 ◽  
Vol 70 (11) ◽  
pp. 6475-6480 ◽  
Author(s):  
Y. P. Tan ◽  
Q. Lin ◽  
X. H. Wang ◽  
S. Joshi ◽  
C. L. Hew ◽  
...  
Keyword(s):  
Dna Sequencing ◽  
Virulence Factors ◽  
Proteomic Analysis ◽  
Virulent Strain ◽  
Pcr Amplification ◽  
Extracellular Proteins ◽  
Edwardsiella Tarda ◽  
Comparative Proteomic Analysis ◽  
Proteomic Approach ◽  
Specific Proteins

ABSTRACT A comparison of extracellular proteins of virulent and avirulent Edwardsiella tarda strains revealed several major, virulent-strain-specific proteins. Proteomic analysis identified two of the proteins in the virulent strain PPD130/91 as flagellin and SseB, which are virulence factors in bacterial pathogens. PCR amplification and DNA sequencing confirmed the presence of the genes that encode these proteins. Our results clearly demonstrated the potency of the proteomic approach in identifying virulence factors.

scholarly journals Clathrin Is Important for Virulence Factors Delivery in the Necrotrophic Fungus Botrytis cinerea

2021 ◽  
Vol 12 ◽  
Author(s):  
Eytham Souibgui ◽  
Christophe Bruel ◽  
Mathias Choquer ◽  
Amélie de Vallée ◽  
Cindy Dieryckx ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Virulence Factors ◽  
Essential Role ◽  
Plant Pathogens ◽  
Pathogenic Fungus ◽  
Hydrolytic Enzymes ◽  
Pathogenic Fungi ◽  
Dominant Negative ◽  
Intracellular Vesicles

Fungi are the most prevalent plant pathogens, causing annually important damages. To infect and colonize their hosts, they secrete effectors including hydrolytic enzymes able to kill and macerate plant tissues. These secreted proteins are transported from the Endoplasmic Reticulum and the Golgi apparatus to the extracellular space through intracellular vesicles. In pathogenic fungi, intracellular vesicles were described but their biogenesis and their role in virulence remain unclear. In this study, we report the essential role of clathrin heavy chain (CHC) in the pathogenicity of Botrytis cinerea, the agent of gray mold disease. To investigate the importance of this protein involved in coat vesicles formation in eukaryotic cells, a T-DNA insertional mutant reduced in the expression of the CHC-encoding gene, and a mutant expressing a dominant-negative form of CHC were studied. Both mutants were strongly affected in pathogenicity. Characterization of the mutants revealed altered infection cushions and an important defect in protein secretion. This study demonstrates the essential role of clathrin in the infectious process of a plant pathogenic fungus and more particularly its role in virulence factors delivery.

scholarly journals Proteomic analysis of mycelium and secretome of different Botrytis cinerea wild-type strains

2014 ◽  
Vol 97 ◽  
pp. 195-221 ◽  
Author(s):  
Raquel González-Fernández ◽  
Kerman Aloria ◽  
José Valero-Galván ◽  
Inmaculada Redondo ◽  
Jesús M Arizmendi ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
Proteomic Analysis ◽  
Wild Type ◽  
Type Strains

Proteomic Analysis of Leaves of the Chlorophyll-Deficient Wheat Mutant Mt6172 and Its Wild-Type through 2D-Difference Gel Electrophoresis

2013 ◽  
Vol 38 (9) ◽  
pp. 1592-1606 ◽  
Author(s):  
Su-Jie SONG ◽  
Jia-Yu GU ◽  
Hui-Jun GUO ◽  
Lin-Shu ZHAO ◽  
Shi-Rong ZHAO ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Gel Electrophoresis ◽  
Wild Type ◽  
Difference Gel Electrophoresis ◽  
Wheat Mutant

A Proteomic Analysis of the Virulence Factors of Three Common Bacterial Species Involved in Periodontitis and Consequent Possible Atherosclerosis: A Narrative Review

2018 ◽  
Vol 19 (11) ◽  
pp. 1124-1130 ◽  
Author(s):  
Reza Mirnejad ◽  
Iman Razeghian-Jahromi ◽  
Masood Sepehrimanesh ◽  
Mohammad Javad Zibaeenezhad ◽  
Pia Lopez-Jornet
Keyword(s):  
Virulence Factors ◽  
Proteomic Analysis ◽  
Bacterial Species ◽  
Narrative Review

An ER–Golgi Tethering Factor SLOH4/MIP3 Is Involved in Long-term Heat Tolerance of Arabidopsis

2020 ◽  
Author(s):  
Kazuho Isono ◽  
Ryo Tsukimoto ◽  
Satoshi Iuchi ◽  
Akihisa Shinozawa ◽  
Izumi Yotsui ◽  
...  
Keyword(s):  
Heat Stress ◽  
Heat Tolerance ◽  
Secretory Proteins ◽  
Wild Type ◽  
Additional Function ◽  
Transcript Levels ◽  
Unfolded Protein ◽  
In The Wild ◽  
Protein Response

Abstract Plants are often exposed not only to short-term (S-) heat stress but also to diurnal long-term (L-) heat stress over several consecutive days. To reveal the mechanisms underlying L-heat stress tolerance, we here used a forward genetic screening for sensitive to long-term heat (sloh) mutants and isolated sloh4. The mutant was hypersensitive to L- but not S-heat stress. The causal gene of sloh4 was identical to MIP3 encoding a member of the MAIGO2 (MAG2) tethering complex, which is composed of the MAG2, MIP1, MIP2, and MIP3 subunits and is localized at the endoplasmic reticulum (ER) membrane. Although sloh4/mip3 was hypersensitive to L-heat stress, the sensitivity of the mag2-3 and mip1–1 mutants was similar to that of the wild type. Under L-heat stress, the ER stress and the following unfolded protein response (UPR) were more pronounced in sloh4 than in the wild type. Transcript levels of bZIP60-regulated UPR genes were strongly increased in sloh4 under L-heat stress. Two processes known to be mediated by INOSITOL REQUIRING ENZYME1 (IRE1)—accumulation of the spliced bZIP60 transcript and a decrease in the transcript levels of PR4 and PRX34, encoding secretory proteins—were observed in sloh4 in response to L-heat stress. These findings suggest that misfolded proteins generated in sloh4 under L-heat stress may be recognized by IRE1 but not bZIP28, resulting in initiation of the UPR via activated bZIP60. Therefore, it would be possible that only MIP3 in MAG2 complex has an additional function in L-heat tolerance, which is not related to the ER–Golgi vesicle tethering.

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