Unraveling the Molecular Mechanisms of Tomatoes’ Defense against Botrytis cinerea: Insights from Transcriptome Analysis of Micro-Tom and Regular Tomato Varieties

Botrytis cinerea is a devastating fungal pathogen that causes severe economic losses in global tomato cultivation. Understanding the molecular mechanisms driving tomatoes’ response to this pathogen is crucial for developing effective strategies to counter it. Although the Micro-Tom (MT) cultivar has been used as a model, its stage-specific response to B. cinerea remains poorly understood. In this study, we examined the response of the MT and Ailsa Craig (AC) cultivars to B. cinerea at different time points (12–48 h post-infection (hpi)). Our results indicated that MT exhibited a stronger resistant phenotype at 18–24 hpi but became more susceptible to B. cinerea later (26–48 hpi) compared to AC. Transcriptome analysis revealed differential gene expression between MT at 24 hpi and AC at 22 hpi, with MT showing a greater number of differentially expressed genes (DEGs). Pathway and functional annotation analysis revealed significant differential gene expression in processes related to metabolism, biological regulation, detoxification, photosynthesis, and carbon metabolism, as well as some immune system-related genes. MT demonstrated an increased reliance on Ca2+ pathway-related proteins, such as CNGCs, CDPKs, and CaMCMLs, to resist B. cinerea invasion. B. cinerea infection induced the activation of PTI, ETI, and SA signaling pathways, involving the modulation of various genes such as FLS2, BAK1, CERK1, RPM, SGT1, and EDS1. Furthermore, transcription factors such as WRKY, MYB, NAC, and AUX/IAA families played crucial regulatory roles in tomatoes’ defense against B. cinerea. These findings provide valuable insights into the molecular mechanisms underlying tomatoes’ defense against B. cinerea and offer potential strategies to enhance plant resistance.

[1]  Xuewen Wang,et al.  Transcriptomic and proteomic analysis reveals (E)-2-hexenal modulates tomato resistance against Botrytis cinerea by regulating plant defense mechanism , 2023, Plant Molecular Biology.

[2]  W. Wang,et al.  Identification of Mycoparasitism-Related Genes against the Phytopathogen Botrytis cinerea via Transcriptome Analysis of Trichoderma harzianum T4 , 2023, Journal of fungi.

[3]  L. Pizarro,et al.  The LeEIX locus determines pathogen resistance in tomato. , 2022, Phytopathology.

[4]  Shaojun Dai,et al.  Pto Interaction Proteins: Critical Regulators in Plant Development and Stress Response , 2022, Frontiers in Plant Science.

[5]  C. Varanda,et al.  Defense Strategies: The Role of Transcription Factors in Tomato–Pathogen Interaction , 2022, Biology.

[6]  Peiguo Yuan,et al.  Calcium/Calmodulin-Mediated Defense Signaling: What Is Looming on the Horizon for AtSR1/CAMTA3-Mediated Signaling in Plant Immunity , 2022, Frontiers in Plant Science.

[7]  G. Pandey,et al.  Calcium signaling and transport machinery: Potential for development of stress tolerance in plants , 2022, Current Plant Biology.

[8]  G. Tena,et al.  PTI and ETI are one , 2021, Nature Plants.

[9]  Ping Wang,et al.  The Glutamate Receptor Plays a Role in Defense against Botrytis cinerea through Electrical Signaling in Tomato , 2021, Applied Sciences.

[10]  T. Odintsova,et al.  Transcriptomic Analysis of Genes Involved in Plant Defense Response to the Cucumber Green Mottle Mosaic Virus Infection , 2021, Life.

[11]  Guo‐Liang Wang,et al.  Ca2+ sensor-mediated ROS scavenging suppresses rice immunity and is exploited by a fungal effector , 2021, Cell.

[12]  K. Harter,et al.  The EDS1–PAD4–ADR1 node mediates Arabidopsis pattern-triggered immunity , 2021, Nature.

[13]  Kunzhi Li,et al.  14-3-3 proteins: regulators of plant metabolism and stress responses. , 2021, Plant biology.

[14]  Xiu-Fang Xin,et al.  PTI-ETI crosstalk: an integrative view of plant immunity. , 2021, Current opinion in plant biology.

[15]  Yuelin Zhang,et al.  Transcriptome analysis reveals that SlNPR1 mediates tomato fruit resistance against Botrytis cinerea by modulating phenylpropanoid metabolism and balancing ROS homeostasis , 2021 .

[16]  J. Parker,et al.  A mis-regulated cyclic nucleotide-gated channel mediates cytosolic calcium elevation and activates immunity in Arabidopsis. , 2021, The New phytologist.

[17]  Y. Liu,et al.  Overexpression of Pti4, Pti5, and Pti6 in tomato promote plant defense and fruit ripening. , 2021, Plant science : an international journal of experimental plant biology.

[18]  I. A. Seman,et al.  CONTROL OF GRAY MOLD DISEASE OF TOMATO CAUSED BY Botrytis cinerea USING BACTERIAL SECONDARY METABOLITES , 2020, Malaysian Applied Biology.

[19]  Diqiu Yu,et al.  The transcription factor WRKY75 positively regulates jasmonate-mediated plant defense to necrotrophic fungal pathogens , 2020, Journal of experimental botany.

[20]  W. Liang,et al.  Systematic Analysis of Lysine Lactylation in the Plant Fungal Pathogen Botrytis cinerea , 2020, Frontiers in Microbiology.

[21]  Guo‐Liang Wang,et al.  Fine-Tuning of RBOH-Mediated ROS Signaling in Plant Immunity. , 2020, Trends in plant science.

[22]  L. Pizarro,et al.  Cytokinin response induces immunity and fungal pathogen resistance, and modulates trafficking of the PRR LeEIX2 in tomato , 2020, Molecular plant pathology.

[23]  A. Sanz,et al.  The Botrytis cinerea Crh1 transglycosylase is a cytoplasmic effector triggering plant cell death and defense response , 2020, Nature Communications.

[24]  Min Ren,et al.  RcMYB84 and RcMYB123 mediate jasmonate-induced defense responses against Botrytis cinerea in rose (Rosa chinensis). , 2020, The Plant journal : for cell and molecular biology.

[25]  Jie Wang,et al.  A full-length transcriptome dataset of normal and Nosema ceranae-challenged midgut tissues of eastern honeybee workers , 2020, bioRxiv.

[26]  Y. Tsang,et al.  Effect of Phenolic Acids Derived from Rice Straw on Botrytis cinerea and Infection on Tomato , 2020, Waste and Biomass Valorization.

[27]  Dayong Li,et al.  NAC transcription factors in plant immunity , 2019, Phytopathology Research.

[28]  Jian Li,et al.  SlMYC2 are required for methyl jasmonate-induced tomato fruit resistance to Botrytis cinerea. , 2019, Food chemistry.

[29]  Zhengbin Zhang,et al.  Transcription Factors Associated with Abiotic and Biotic Stress Tolerance and Their Potential for Crops Improvement , 2019, Genes.

[30]  S. Steinkellner,et al.  Serendipita Species Trigger Cultivar-Specific Responses to Fusarium Wilt in Tomato , 2019, Agronomy.

[31]  A. Si-Ammour,et al.  'Candidatus Phytoplasma mali' genome encodes a protein that functions as a E3 Ubiquitin Ligase and could inhibit plant basal defense. , 2019, Molecular plant-microbe interactions : MPMI.

[32]  Xinghong Yang,et al.  Genetic engineering of the biosynthesis of glycinebetaine enhances the fruit development and size of tomato. , 2019, Plant science : an international journal of experimental plant biology.

[33]  W. Moeder,et al.  Ca2+ to the rescue - Ca2+channels and signaling in plant immunity. , 2019, Plant science : an international journal of experimental plant biology.

[34]  D. W. Ng,et al.  Regulating the Regulators: The Control of Transcription Factors in Plant Defense Signaling , 2018, International journal of molecular sciences.

[35]  J. Ragoussis,et al.  Transcriptome landscape of the developing olive fruit fly embryo delineated by Oxford Nanopore long-read RNA-Seq , 2018, bioRxiv.

[36]  Wenqing Yu,et al.  SlERF2 Is Associated with Methyl Jasmonate-Mediated Defense Response against Botrytis cinerea in Tomato Fruit. , 2018, Journal of agricultural and food chemistry.

[37]  J. V. van Kan,et al.  Many Shades of Grey in Botrytis-Host Plant Interactions. , 2018, Trends in plant science.

[38]  Suk-Young Hong,et al.  Genome-wide transcriptomic analysis of BR-deficient Micro-Tom reveals correlations between drought stress tolerance and brassinosteroid signaling in tomato. , 2018, Plant physiology and biochemistry : PPB.

[39]  A. Fisher,et al.  The Receptor-like Cytoplasmic Kinase BIK1 Localizes to the Nucleus and Regulates Defense Hormone Expression during Plant Innate Immunity. , 2018, Cell host & microbe.

[40]  Xuefeng Ma,et al.  A Verticillium dahliae Extracellular Cutinase Modulates Plant Immune Responses. , 2018, Molecular plant-microbe interactions : MPMI.

[41]  I. Nookaew,et al.  Complete genomic and transcriptional landscape analysis using third-generation sequencing: a case study of Saccharomyces cerevisiae CEN.PK113-7D , 2018, Nucleic acids research.

[42]  C. Gaillard,et al.  The molecular dialogue between Arabidopsis thaliana and the necrotrophic fungus Botrytis cinerea leads to major changes in host carbon metabolism , 2017, Scientific Reports.

[43]  O. Rupp,et al.  Pathogen recognition in compatible plant-microbe interactions , 2017, Scientific Reports.

[44]  Itai Sharon,et al.  BcXYG1, a Secreted Xyloglucanase from Botrytis cinerea, Triggers Both Cell Death and Plant Immune Responses1 , 2017, Plant Physiology.

[45]  W. Guo,et al.  Verticillium dahliae manipulates plant immunity by glycoside hydrolase 12 proteins in conjunction with carbohydrate‐binding module 1 , 2017, Environmental microbiology.

[46]  João P. Bezerra Neto,et al.  Transcription Factors Involved in Plant Resistance to Pathogens. , 2017, Current protein & peptide science.

[47]  Sanwen Huang,et al.  A chemical genetic roadmap to improved tomato flavor , 2017, Science.

[48]  Synan F. AbuQamar,et al.  Mechanisms and strategies of plant defense against Botrytis cinerea , 2017, Critical reviews in biotechnology.

[49]  G. Martin,et al.  iTAK: A Program for Genome-wide Prediction and Classification of Plant Transcription Factors, Transcriptional Regulators, and Protein Kinases. , 2016, Molecular plant.

[50]  Xin-Zhong Cai,et al.  Calcium-dependent protein kinase (CDPK) and CDPK-related kinase (CRK) gene families in tomato: genome-wide identification and functional analyses in disease resistance , 2016, Molecular Genetics and Genomics.

[51]  Doughari Jh,et al.  An Overview of Plant Immunity , 2015 .

[52]  Tomás C. Moyano,et al.  Transcriptome analysis reveals regulatory networks underlying differential susceptibility to Botrytis cinerea in response to nitrogen availability in Solanum lycopersicum , 2015, Front. Plant Sci..

[53]  Xin-Zhong Cai,et al.  Calcium-dependent protein kinase (CDPK) and CDPK-related kinase (CRK) gene families in tomato: genome-wide identification and functional analyses in disease resistance , 2015, Molecular Genetics and Genomics.

[54]  Yang Wang,et al.  A Phytophthora sojae Glycoside Hydrolase 12 Protein Is a Major Virulence Factor during Soybean Infection and Is Recognized as a PAMP[OPEN] , 2015, Plant Cell.

[55]  K. Ohnishi,et al.  Molecular chaperons and co-chaperons, Hsp90, RAR1, and SGT1 negatively regulate bacterial wilt disease caused by Ralstonia solanacearum in Nicotiana benthamiana , 2015, Plant signaling & behavior.

[56]  Jie Zhou,et al.  Characterization of the promoter and extended C-terminal domain of Arabidopsis WRKY33 and functional analysis of tomato WRKY33 homologues in plant stress responses , 2015, Journal of experimental botany.

[57]  Xin-Zhong Cai,et al.  Cyclic nucleotide gated channel gene family in tomato: genome-wide identification and functional analyses in disease resistance , 2015, Front. Plant Sci..

[58]  Imre E Somssich,et al.  Transcriptional networks in plant immunity. , 2015, The New phytologist.

[59]  D. Rav-David,et al.  Induced systemic resistance in tomato (Solanum lycopersicum) against Botrytis cinerea by biochar amendment involves jasmonic acid signaling , 2015, Plant and Soil.

[60]  Dayong Li,et al.  Tomato NAC Transcription Factor SlSRN1 Positively Regulates Defense Response against Biotic Stress but Negatively Regulates Abiotic Stress Response , 2014, PloS one.

[61]  T. Mengiste,et al.  Resistance to Botrytis cinerea in Solanum lycopersicoides involves widespread transcriptional reprogramming , 2014, BMC Genomics.

[62]  L. G. Fietto,et al.  Transcription Factor Functional Protein-Protein Interactions in Plant Defense Responses , 2014, Proteomes.

[63]  D. Rav-David,et al.  Systemic resistance to gray mold induced in tomato by benzothiadiazole and Trichoderma harzianum T39. , 2014, Phytopathology.

[64]  Mark D. Robinson,et al.  Robustly detecting differential expression in RNA sequencing data using observation weights , 2013, Nucleic acids research.

[65]  Damian Szklarczyk,et al.  eggNOG v4.0: nested orthology inference across 3686 organisms , 2013, Nucleic Acids Res..

[66]  C. Clément,et al.  Grapevine NAC1 transcription factor as a convergent node in developmental processes, abiotic stresses, and necrotrophic/biotrophic pathogen tolerance. , 2013, Journal of experimental botany.

[67]  A. Powell,et al.  Tomato transcriptome and mutant analyses suggest a role for plant stress hormones in the interaction between fruit and Botrytis cinerea , 2013, Front. Plant Sci..

[68]  S. Eddy,et al.  Challenges in homology search: HMMER3 and convergent evolution of coiled-coil regions , 2013, Nucleic acids research.

[69]  B. Fox,et al.  Mutations in FLS2 Ser-938 Dissect Signaling Activation in FLS2-Mediated Arabidopsis Immunity , 2013, PLoS pathogens.

[70]  Karsten M. Borgwardt,et al.  Arabidopsis Defense against Botrytis cinerea: Chronology and Regulation Deciphered by High-Resolution Temporal Transcriptomic Analysis[C][W][OA] , 2012, Plant Cell.

[71]  C. Beuzón,et al.  CML9, an Arabidopsis calmodulin-like protein, contributes to plant innate immunity through a flagellin-dependent signalling pathway. , 2012, The Plant journal : for cell and molecular biology.

[72]  C. Zipfel,et al.  Plant pattern recognition receptor complexes at the plasma membrane. , 2012, Current opinion in plant biology.

[73]  A. Powell,et al.  Proteomic analysis of ripening tomato fruit infected by Botrytis cinerea. , 2012, Journal of proteome research.

[74]  Rainer P Birkenbihl,et al.  Arabidopsis WRKY33 Is a Key Transcriptional Regulator of Hormonal and Metabolic Responses toward Botrytis cinerea Infection1[W] , 2012, Plant Physiology.

[75]  Hui Liu,et al.  AnimalTFDB: a comprehensive animal transcription factor database , 2011, Nucleic Acids Res..

[76]  F. Daayf,et al.  Botrytis cinerea Manipulates the Antagonistic Effects between Immune Pathways to Promote Disease Development in Tomato[C][W][OA] , 2011, Plant Cell.

[77]  W. Huber,et al.  which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. MAnorm: a robust model for quantitative comparison of ChIP-Seq data sets , 2011 .

[78]  Vladimir B Bajic,et al.  Transcriptional regulatory network triggered by oxidative signals configures the early response mechanisms of japonica rice to chilling stress , 2010, BMC Plant Biology.

[79]  V. Flors,et al.  Hexanoic acid-induced resistance against Botrytis cinerea in tomato plants. , 2009, Molecular plant-microbe interactions : MPMI.

[80]  Fengming Song,et al.  The Arabidopsis ATAF1, a NAC transcription factor, is a negative regulator of defense responses against necrotrophic fungal and bacterial pathogens. , 2009, Molecular plant-microbe interactions : MPMI.

[81]  I. Somssich,et al.  The Role of WRKY Transcription Factors in Plant Immunity[W] , 2009, Plant Physiology.

[82]  M. Bolton Primary metabolism and plant defense--fuel for the fire. , 2009, Molecular plant-microbe interactions : MPMI.

[83]  B. Thomma,et al.  Genetic Dissection of Verticillium Wilt Resistance Mediated by Tomato Ve11[C][W][OA] , 2009, Plant Physiology.

[84]  Synan F. AbuQamar,et al.  Tomato Protein Kinase 1b Mediates Signaling of Plant Responses to Necrotrophic Fungi and Insect Herbivory[W] , 2008, The Plant Cell Online.

[85]  M. Rep,et al.  Suppression of Plant Resistance Gene-Based Immunity by a Fungal Effector , 2008, PLoS pathogens.

[86]  C. Zipfel Pattern-recognition receptors in plant innate immunity. , 2008, Current opinion in immunology.

[87]  E. Birney,et al.  Pfam: the protein families database , 2013, Nucleic Acids Res..

[88]  Brian Williamson,et al.  Botrytis cinerea: the cause of grey mould disease. , 2007, Molecular plant pathology.

[89]  Peter van Baarlen,et al.  Molecular mechanisms of pathogenicity: how do pathogenic microorganisms develop cross-kingdom host jumps? , 2007, FEMS microbiology reviews.

[90]  Jonathan D. G. Jones,et al.  The plant immune system , 2006, Nature.

[91]  U. Sonnewald,et al.  Plant-microbe interactions to probe regulation of plant carbon metabolism. , 2006, Journal of plant physiology.

[92]  Xiaoyan Tang,et al.  Flagellin induces innate immunity in nonhost interactions that is suppressed by Pseudomonas syringae effectors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[93]  D. Shibata,et al.  Catalog of Micro-Tom tomato responses to common fungal, bacterial, and viral pathogens , 2005, Journal of General Plant Pathology.

[94]  Jodie J. Yin,et al.  A comprehensive evolutionary classification of proteins encoded in complete eukaryotic genomes , 2004, Genome Biology.

[95]  S. He,et al.  A Pseudomonas syringae type III effector suppresses cell wall-based extracellular defense in susceptible Arabidopsis plants , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[96]  David H. Parker,et al.  Ethylene Production by Botrytis cinerea In Vitro and in Tomatoes , 2002, Applied and Environmental Microbiology.

[97]  Karam B. Singh,et al.  Transcription factors in plant defense and stress responses. , 2002, Current opinion in plant biology.

[98]  M. Höfte,et al.  Abscisic Acid Determines Basal Susceptibility of Tomato toBotrytis cinerea and Suppresses Salicylic Acid-Dependent Signaling Mechanisms1 , 2002, Plant Physiology.

[99]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[100]  Lihuang Zhu,et al.  Overexpression of Pti5 in tomato potentiates pathogen-induced defense gene expression and enhances disease resistance to Pseudomonas syringae pv. tomato. , 2001, Molecular plant-microbe interactions : MPMI.

[101]  G. Martin,et al.  Pti4 Is Induced by Ethylene and Salicylic Acid, and Its Product Is Phosphorylated by the Pto Kinase , 2000, Plant Cell.

[102]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.

[103]  M. Farman,et al.  Chromosome walking to the AVR1-CO39 avirulence gene of Magnaporthe grisea: discrepancy between the physical and genetic maps. , 1998, Genetics.

[104]  R. Hedrich,et al.  Receptor-mediated activation of a plant Ca2+-permeable ion channel involved in pathogen defense. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[105]  L. M. Pérez,et al.  Calcium ions promote the response of Citrus limon against fungal elicitors or wounding , 1996 .

[106]  M. Causse,et al.  Trait discovery and editing in tomato , 2018, The Plant journal : for cell and molecular biology.

[107]  J. Grosser,et al.  Potential use of the DREB/ERF, MYB, NAC and WRKY transcription factors to improve abiotic and biotic stress in transgenic plants , 2017, Plant Cell, Tissue and Organ Culture (PCTOC).

[108]  Linlin Li,et al.  Induction of disease resistance by salicylic acid and calcium ion against Botrytis cinerea in tomato (Lycopersicon esculentum) , 2017 .

[109]  H. Ezura,et al.  Micro-Tom Tomato as an Alternative Plant Model System: Mutant Collection and Efficient Transformation. , 2016, Methods in molecular biology.

[110]  A. E. Hadrami,et al.  Botrytis cinereaManipulates the Antagonistic Effects between Immune Pathways to Promote Disease Development in Tomato , 2011 .

[111]  He Fuchu,et al.  Integrated nr Database in Protein Annotation System and Its Localization , 2006 .

[112]  Susumu Goto,et al.  The KEGG resource for deciphering the genome , 2004, Nucleic Acids Res..

[113]  Michael Y. Galperin,et al.  The COG database: a tool for genome-scale analysis of protein functions and evolution , 2000, Nucleic Acids Res..

[114]  Thomas D. Schmittgen,et al.  Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method , 2022 .