cc72e83733cea5ddbbf65baae5b17aaa 26TV01_MolPlantPathol_SColl_Valls_pub.pdf fefb1100dfdc90f658be3f507101ee4e0600a95d 26TV01_MolPlantPathol_SColl_Valls_pub.pdf 620dd00713edaf03d9d9ed9758ce8967fc234f0f6142d76f981e22b0e00396c4 26TV01_MolPlantPathol_SColl_Valls_pub.pdf Title: The Effector RipAW Enhances Ralstonia solanacearum Invasion in Arabidopsis via CBP60g/SARD1‐Dependent and ‐Independent Pathways Subject: Molecular Plant Pathology 2026.27:e70207 Keywords: Keywords: CBP60g; plant susceptibility; Ralstonia solanacearum; RipAW; SARD1 Author: Huijuan Wang,Shouyang Fu, Tao Cao, Yang Niu, Shengyang Cheng, Qichang Gong, Hui Ma, Xiang Wang, Jinxue Hu, Min Chen, Dongdong Wang, Yong Zhang, Nuria S. Coll, Marc Valls, Qin Chen, Cuizhu Zhao, Yue Chen, Haibin Lu Creator: Adobe InDesign 19.5 (Windows) Producer: Adobe PDF Library 17.0; modified using iTextSharp 5.5.13.4 ©2000-2024 iText Group NV (AGPL-version) CreationDate: Thu Jan 22 05:05:49 2026 CET ModDate: Thu Jan 22 09:52:30 2026 CET Custom 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Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 0 FreeObjects: 2 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: true NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Language: en-US Outlines: Item: Title: The Effector RipAW Enhances Ralstonia solanacearum Invasion in Arabidopsis via CBP60g/SARD1-Dependent and -Independent Pathways Children: Item: Title: ABSTRACT Item: Title: 1   |   Introduction Item: Title: 2   |   Results Children: Item: Title: 2.1   |   Type III Effector RipAW From R. solanacearum GMI1000 Increases Virulence in Arabidopsis Item: Title: 2.2   |   Deletion of RipAW Reduces R. solanacearum Virulence in Arabidopsis Item: Title: 2.3   |   E3 Ligase Activity Is Essential for the RipAW Virulence Function in Plants Item: Title: 2.4   |   Transcriptome Profiling Reveals That CBP60g and SARD1 Upregulation in Est::RipAW Transgenic Plants Does Not Activate the SA Signalling Pathway Item: Title: 2.5   |   RipAW Associates With and Destabilises CBP60g in E3 Ligase Activity-Dependent Way Item: Title: 2.6   |   CBP60g and SARD1 Are Involved in Plant Resistance to RipAW-Mediated R. solanacearum Multiplication in Arabidopsis, but Not in RipAW-Mediated Root Architecture Changes Item: Title: 3   |   Discussion Item: Title: 4   |   Experimental Procedures Children: Item: Title: 4.1   |   Plant Materials and Microbial Strains Item: Title: 4.2   |   Plasmid and Transgenic Plants Generation Item: Title: 4.3   |   Generation of the R. solanacearum Complemented Strain (CΔripAW) Item: Title: 4.4   |   Ralstonia solanacearum Infection Assays Item: Title: 4.5   |   Sample Preparation for RNA Seq and Data Analysis Item: Title: 4.6   |   GO Analysis Item: Title: 4.7   |   RT-qPCR Item: Title: 4.8   |   SA Measurement Item: Title: 4.9   |   Agrobacterium-Mediated Transient Expression on N. benthamiana Item: Title: 4.10   |   Subcellular Localisation, Split-Luciferase Assay and BiFC Assay Item: Title: 4.11   |   Co-IP Assay Item: Title: 4.12   |   Statistical Analysis Item: Title: Author Contributions Item: Title: Acknowledgements Item: Title: Funding Item: Title: Conflicts of Interest Item: Title: Data Availability Statement Item: Title: References Filters: FilterPipeline: FlateDecode FilterPipeline: CCITTFaxDecode FilterPipeline: DCTDecode Images: Image: NisoImageMetadata: FormatName: image/png ImageWidth: 228 ImageHeight: 45 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Image: NisoImageMetadata: FormatName: image/png ImageWidth: 126 ImageHeight: 126 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Filter: FlateDecode Intent: RelativeColorimetric Name: X Image: NisoImageMetadata: FormatName: image/tiff 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Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 101, 561, 104, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 166, 561, 169, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 212, 561, 215, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 262, 561, 265, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 346, 561, 349, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 416, 561, 419, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 460, 561, 463, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 521, 561, 525, 548 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 85, 550, 88, 537 ActionDest: 297 Annotation: Subtype: Link Contents: College of Resources and EnvironmentSouthwest UniversityChongqingChina Rect: 138, 550, 146, 537 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 222, 550, 225, 537 ActionDest: 297 Annotation: Subtype: Link Contents: College of Resources and EnvironmentSouthwest UniversityChongqingChina Rect: 283, 550, 291, 537 ActionDest: 297 Annotation: Subtype: Link Contents: Centre for Research in Agricultural Genomics (CSICIRTAUABUB)BellaterraCataloniaSpain Rect: 352, 550, 360, 537 ActionDest: 297 Annotation: Subtype: Link Contents: Centre for Research in Agricultural Genomics (CSICIRTAUABUB)BellaterraCataloniaSpain Rect: 413, 550, 421, 537 ActionDest: 297 Annotation: Subtype: Link Contents: College of Food Science and EngineeringNorthwest A&F UniversityYanglingChina Rect: 470, 550, 473, 537 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 534, 550, 537, 537 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 82, 539, 86, 526 ActionDest: 297 Annotation: Subtype: Link Contents: State Key Laboratory for Crop Stress Resistance and HighEfficiency Production, College of AgronomyNorthwest A&F UniversityYanglingChina Rect: 136, 539, 139, 526 ActionDest: 297 Annotation: Subtype: Link Contents: Please visit https://orcid.org/0000-0002-0613-4697 for your more information Rect: 139, 540, 152, 528 Annotation: Subtype: Link Contents: Please visit mailto:xnchenyue@nwafu.edu.cn for your more information Rect: 143, 453, 229, 442 Annotation: Subtype: Link Contents: Please visit mailto:luhaibin011@hotmail.com for your more information Rect: 279, 453, 365, 442 Annotation: Subtype: Link Contents: Coll, N. 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X.Feng, etal. 2023. “The Ralstonia solanacearum Type III Effector RipAW Targets the Immune Receptor Complex to Suppress PAMPTriggered Immunity.” International Journal of Molecular Sciences25: 183. Rect: 487, 96, 505, 83 ActionDest: 288 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 265, 498, 283, 486 ActionDest: 288 Annotation: Subtype: Link Contents: Figure S1: Plants constitutiveexpressing RipAW exhibit defects in growth. (a) The growth phenotypes of 10weekold Col0 plants and 10weekold 35S::RipAW T1 transgenic lines. Lines 1# and 2# showed sever dwarf phenotype while line 3# is less dwarf. All of them did not produce seeds. (b) Identification of RipAW expression in Est::RipAW transgenic lines. Sixdayold seedlings were transferred on MS2 containing 5M estradiol. At 48hpe, total proteins from seedlings were extracted and performed immunoblot with antiflag antibody for detecting RipAW protein. Rect: 207, 429, 220, 417 ActionDest: 290 Annotation: Subtype: Link Contents: Figure S1: Plants constitutiveexpressing RipAW exhibit defects in growth. (a) The growth phenotypes of 10weekold Col0 plants and 10weekold 35S::RipAW T1 transgenic lines. Lines 1# and 2# showed sever dwarf phenotype while line 3# is less dwarf. All of them did not produce seeds. (b) Identification of RipAW expression in Est::RipAW transgenic lines. Sixdayold seedlings were transferred on MS2 containing 5M estradiol. At 48hpe, total proteins from seedlings were extracted and performed immunoblot with antiflag antibody for detecting RipAW protein. Rect: 77, 372, 90, 359 ActionDest: 290 Annotation: Subtype: Link Contents: Lu, H., A. S.Lema, M.PlanasMarques, A.AlonsoDiaz, M.Valls, and N. S.Coll. 2018. “Type III SecretionDependent and Independent Phenotypes Caused by Ralstonia solanacearum in Arabidopsis Roots.” Molecular Plant Microbe Interactions31: 175 184. Rect: 253, 360, 270, 348 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 1a,b" for your more reference Rect: 268, 314, 283, 302 ActionDest: 129 Annotation: Subtype: Link Contents: Please see "figure 1a,c" for your more reference Rect: 269, 291, 283, 279 ActionDest: 129 Annotation: Subtype: Link Contents: Lu, H., A. S.Lema, M.PlanasMarques, A.AlonsoDiaz, M.Valls, and N. S.Coll. 2018. “Type III SecretionDependent and Independent Phenotypes Caused by Ralstonia solanacearum in Arabidopsis Roots.” Molecular Plant Microbe Interactions31: 175 184. Rect: 85, 268, 103, 256 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 1d" for your more reference Rect: 274, 257, 283, 244 ActionDest: 129 Annotation: Subtype: Link Contents: Lu, H., A. S.Lema, M.PlanasMarques, A.AlonsoDiaz, M.Valls, and N. S.Coll. 2018. “Type III SecretionDependent and Independent Phenotypes Caused by Ralstonia solanacearum in Arabidopsis Roots.” Molecular Plant Microbe Interactions31: 175 184. Rect: 200, 176, 218, 164 ActionDest: 288 Annotation: Subtype: Link Contents: Zhao, C., H.Wang, Y.Lu, etal. 2019. “Deep Sequencing Reveals Early Reprogramming of Arabidopsis Root Transcriptomes Upon Ralstonia solanacearum Infection.” Molecular Plant Microbe Interactions32: 813 827. Rect: 266, 176, 283, 164 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 1e" for your more reference Rect: 398, 751, 406, 739 ActionDest: 129 Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 528, 636, 546, 623 ActionDest: 288 Annotation: Subtype: Link Contents: Figure S2: RipAW plays an important role in R.solanacearum invasion on adult plants. (a) Characterisation of the complemented strain CRipAW genotype by PCR. (b) Detection of RipAW expression in CRipAW strain by qPCR. Gene expression was quantified and normalised to 16S rRNA using the 2−Ct method. (c) The growth of different R.solanacearum strains in rich medium. (d) RipAW is required for R.solanacearum colonisation using a hydroponic infection system (n=6). A 2cm tip fragment of roots from 5weekold plants grown in Arabidopsis nutrient solution (ANS) solution was cut off. Plants were then transferred into suspensions of R.solanacearum of the indicated strains (OD600=0.1). Bacterial populations were measured in the aerial parts of the plants at 5 dpi. Statistical analysis was done with oneway ANOVA Tukey's test (*p<0.05). The experiment was performed twice with similar results. Rect: 337, 625, 359, 612 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 2a c" for your more reference Rect: 469, 579, 486, 566 ActionDest: 146 Annotation: Subtype: Link Contents: Please see "figure 2d" for your more reference Rect: 475, 544, 485, 531 ActionDest: 146 Annotation: Subtype: Link Contents: Please see "figure 2e" for your more reference Rect: 411, 498, 419, 485 ActionDest: 146 Annotation: Subtype: Link Contents: Figure S2: RipAW plays an important role in R.solanacearum invasion on adult plants. (a) Characterisation of the complemented strain CRipAW genotype by PCR. (b) Detection of RipAW expression in CRipAW strain by qPCR. Gene expression was quantified and normalised to 16S rRNA using the 2−Ct method. (c) The growth of different R.solanacearum strains in rich medium. (d) RipAW is required for R.solanacearum colonisation using a hydroponic infection system (n=6). A 2cm tip fragment of roots from 5weekold plants grown in Arabidopsis nutrient solution (ANS) solution was cut off. Plants were then transferred into suspensions of R.solanacearum of the indicated strains (OD600=0.1). Bacterial populations were measured in the aerial parts of the plants at 5 dpi. Statistical analysis was done with oneway ANOVA Tukey's test (*p<0.05). The experiment was performed twice with similar results. Rect: 530, 475, 544, 462 ActionDest: 290 Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 367, 360, 385, 347 ActionDest: 288 Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 483, 348, 501, 335 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 3a" for your more reference Rect: 419, 291, 427, 278 ActionDest: 162 Annotation: Subtype: Link Contents: Please see "figure 3b" for your more reference Rect: 492, 279, 501, 266 ActionDest: 162 Annotation: Subtype: Link Contents: Please see "figure 3c" for your more reference Rect: 388, 199, 396, 186 ActionDest: 162 Page: Sequence: 4 Annotations: Annotation: Subtype: Link Contents: Please see "figure 1" for your more reference Rect: 246, 222, 250, 209 ActionDest: 129 Annotation: Subtype: Link Contents: Please see "figure 4a" for your more reference Rect: 218, 164, 227, 151 ActionDest: 176 Annotation: Subtype: Link Contents: Please see "figure 4a" for your more reference Rect: 180, 141, 189, 128 ActionDest: 176 Annotation: Subtype: Link Contents: Data S1: Differentially expressed genes in roots of RipAW transgenic plants and WT plants. Rect: 214, 141, 223, 128 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 4a" for your more reference Rect: 508, 222, 516, 209 ActionDest: 176 Annotation: Subtype: Link Contents: Please see "figure 4a" for your more reference Rect: 337, 153, 345, 140 ActionDest: 176 Annotation: Subtype: Link Contents: Sun, T., Y.Zhang, Y.Li, Q.Zhang, and Y.Ding. 2015. “ChIPSeq Reveals Broad Roles of SARD1 and CBP60g in Regulating Plant Immunity.” Nature Communications6: 10159. Rect: 527, 118, 544, 105 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "table 1" for your more reference Rect: 332, 95, 336, 82 ActionDest: 180 Annotation: Subtype: Link Contents: TableS2: The expression of SA biosynthesis and signalling pathway genes in RNAseq data. Rect: 379, 95, 388, 82 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 4b" for your more reference Rect: 380, 61, 389, 48 ActionDest: 176 Annotation: Subtype: Link Contents: Figure S3: Identification the expressions of CBP60g and SARD1 in Est::RipAW transgenic line 16#. Genes expressions of CBP60g and SARD1 in Est::RipAW transgenic 16# seedlings were measured by RTqPCR at 48hpe. Gene expression was quantified and normalised to AtActin2 using the 2−Ct method. The experiment was performed three with similar results. Statistical analysis was done with oneway ANOVA Sidka's test (**p<0.01, ***p<0.001). Rect: 436, 61, 445, 48 ActionDest: 290 Page: Sequence: 5 Annotations: Annotation: Subtype: Link Contents: Zhang, Y., S.Xu, P.Ding, etal. 2010. “Control of Salicylic Acid Synthesis and Systemic Acquired Resistance by Two Members of a PlantSpecific Family of Transcription Factors.” Proceedings of the National Academy of Sciences of the United States of America107: 18220 18225. Rect: 266, 360, 283, 347 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "table 1" for your more reference Rect: 113, 279, 117, 266 ActionDest: 180 Annotation: Subtype: Link Contents: TableS2: The expression of SA biosynthesis and signalling pathway genes in RNAseq data. Rect: 159, 279, 168, 266 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 4b" for your more reference Rect: 208, 233, 216, 220 ActionDest: 176 Annotation: Subtype: Link Contents: Please see "figure 4c" for your more reference Rect: 148, 199, 156, 186 ActionDest: 176 Annotation: Subtype: Link Contents: Please see "figure 5a,b" for your more reference Rect: 438, 302, 454, 289 ActionDest: 194 Annotation: Subtype: Link Contents: Figure S4: RipAW specifically affects CBP60g stablility. (a) RipAW promoted CBP60g degradation when they were coexpressed in N.benthamiana. (b) RipAW could not trigger GFPHAnLUC degradation when they were coexpressed in N.benthamiana. The indicated genes were transiently expressed in N.benthamiana by agrobacteriummediated transformation. At 1 dpi, the agroinfiltrated leaves were treated with CHX for 4h. The samples were collected and the indicated proteins were tested by western blot. Rect: 499, 302, 519, 289 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 5c,d" for your more reference Rect: 371, 245, 387, 232 ActionDest: 194 Annotation: Subtype: Link Contents: Please see "figure 5c,d" for your more reference Rect: 336, 210, 351, 197 ActionDest: 194 Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 395, 141, 414, 128 ActionDest: 288 Annotation: Subtype: Link Contents: Ouyang, X., J.Chen, Z.Sun, etal. 2023. “Ubiquitin E3 Ligase Activity of Ralstonia solanacearum Effector RipAW Is Not Essential for Induction of Plant Defense in Nicotiana benthamiana.” Frontiers in Microbiology14: 1201444. Rect: 477, 141, 495, 128 ActionDest: 288 Annotation: Subtype: Link Contents: Figure S5: Subcellular localisation of RipAW (C177S). RipAW (C177S)GFP was transiently expressed on N.benthamiana by agroinfiltration. At 2dpi, subcellular localisation of RipAW (C177S)GFP were digitally photographed by a laserscanning confocal microscope (Zeiss LSM880), mCherry carrying with NLS was used to visualise nucleus, bar=50m. The proteins were detected by western blot with antiGFP antibody. Rect: 478, 72, 487, 59 ActionDest: 290 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Contents: Sun, Z. M., Q.Zhang, Y. X.Feng, etal. 2023. “The Ralstonia solanacearum Type III Effector RipAW Targets the Immune Receptor Complex to Suppress PAMPTriggered Immunity.” International Journal of Molecular Sciences25: 183. Rect: 393, 559, 411, 547 ActionDest: 288 Annotation: Subtype: Link Contents: Figure S5: Subcellular localisation of RipAW (C177S). RipAW (C177S)GFP was transiently expressed on N.benthamiana by agroinfiltration. At 2dpi, subcellular localisation of RipAW (C177S)GFP were digitally photographed by a laserscanning confocal microscope (Zeiss LSM880), mCherry carrying with NLS was used to visualise nucleus, bar=50m. The proteins were detected by western blot with antiGFP antibody. Rect: 449, 559, 458, 547 ActionDest: 290 Annotation: Subtype: Link Contents: Figure S5: Subcellular localisation of RipAW (C177S). RipAW (C177S)GFP was transiently expressed on N.benthamiana by agroinfiltration. At 2dpi, subcellular localisation of RipAW (C177S)GFP were digitally photographed by a laserscanning confocal microscope (Zeiss LSM880), mCherry carrying with NLS was used to visualise nucleus, bar=50m. The proteins were detected by western blot with antiGFP antibody. Rect: 510, 536, 519, 524 ActionDest: 290 Annotation: Subtype: Link Contents: Celenza, J. L., J. A.Quiel, G. A.Smolen, etal. 2005. “The Arabidopsis ATR1 Myb Transcription Factor Controls Indolic Glucosinolate Homeostasis.” Plant Physiology137: 253 262. Rect: 329, 410, 348, 397 ActionDest: 284 Annotation: Subtype: Link Contents: Wang, Y., A.Zhao, R. J. L.Morcillo, etal. 2021. “A Bacterial Effector Protein Uncovers a Plant Metabolic Pathway Involved in Tolerance to Bacterial Wilt Disease.” Molecular Plant14: 1281 1296. Rect: 402, 410, 420, 397 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 5e" for your more reference Rect: 392, 375, 401, 363 ActionDest: 194 Annotation: Subtype: Link Contents: Please see "figure 5f" for your more reference Rect: 367, 352, 375, 340 ActionDest: 194 Annotation: Subtype: Link Contents: Please see "figure 5g" for your more reference Rect: 363, 329, 372, 317 ActionDest: 194 Annotation: Subtype: Link Contents: Please see "figure 6a d" for your more reference Rect: 437, 135, 456, 122 ActionDest: 213 Page: Sequence: 7 Page: Sequence: 8 Annotations: Annotation: Subtype: Link Contents: Please see "figure 6e" for your more reference Rect: 145, 245, 153, 232 ActionDest: 213 Annotation: Subtype: Link Contents: Please see "figure 6f h" for your more reference Rect: 212, 222, 228, 209 ActionDest: 213 Annotation: Subtype: Link Contents: Please see "figure 6e" for your more reference Rect: 178, 95, 187, 82 ActionDest: 213 Annotation: Subtype: Link Contents: Please see "figure 7a c" for your more reference Rect: 363, 176, 380, 163 ActionDest: 222 Annotation: Subtype: Link Contents: Please see "figure 7d" for your more reference Rect: 363, 118, 372, 105 ActionDest: 222 Annotation: Subtype: Link Contents: Please refer footnote number a Rect: 86, 721, 89, 708 ActionDest: 180 Annotation: Subtype: Link Contents: Please refer footnote number b Rect: 521, 721, 525, 708 ActionDest: 180 Page: Sequence: 9 Page: Sequence: 10 Annotations: Annotation: Subtype: Link Contents: BundalovicTorma, C., F.Lonjon, D.Desveaux, and D. S.Guttman. 2022. “Diversity, Evolution, and Function of Pseudomonas syringae Effectoromes.” Annual Review of Phytopathology60: 211 236. Rect: 187, 417, 206, 404 ActionDest: 284 Annotation: Subtype: Link Contents: BundalovicTorma, C., F.Lonjon, D.Desveaux, and D. S.Guttman. 2022. “Diversity, Evolution, and Function of Pseudomonas syringae Effectoromes.” Annual Review of Phytopathology60: 211 236. Rect: 265, 383, 283, 370 ActionDest: 284 Annotation: Subtype: Link Contents: Peng, Y., J.Yang, X.Li, and Y.Zhang. 2021. “Salicylic Acid: Biosynthesis and Signaling.” Annual Review of Plant Biology72: 761 791. Rect: 128, 337, 147, 324 ActionDest: 288 Annotation: Subtype: Link Contents: Sun, T., Y.Zhang, Y.Li, Q.Zhang, and Y.Ding. 2015. “ChIPSeq Reveals Broad Roles of SARD1 and CBP60g in Regulating Plant Immunity.” Nature Communications6: 10159. Rect: 194, 337, 212, 324 ActionDest: 288 Annotation: Subtype: Link Contents: Qin, J., K.Wang, L.Sun, etal. 2018. “The PlantSpecific Transcription Factors CBP60g and SARD1 Are Targeted by a Verticillium Secretory Protein VdSCP41 to Modulate Immunity.” eLife7: e34902. Rect: 265, 118, 283, 105 ActionDest: 288 Annotation: Subtype: Link Contents: Qin, J., K.Wang, L.Sun, etal. 2018. “The PlantSpecific Transcription Factors CBP60g and SARD1 Are Targeted by a Verticillium Secretory Protein VdSCP41 to Modulate Immunity.” eLife7: e34902. Rect: 265, 72, 283, 59 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 1" for your more reference Rect: 400, 452, 404, 439 ActionDest: 129 Annotation: Subtype: Link Contents: Lu, H., A. S.Lema, M.PlanasMarques, A.AlonsoDiaz, M.Valls, and N. S.Coll. 2018. “Type III SecretionDependent and Independent Phenotypes Caused by Ralstonia solanacearum in Arabidopsis Roots.” Molecular Plant Microbe Interactions31: 175 184. Rect: 370, 429, 388, 416 ActionDest: 288 Annotation: Subtype: Link Contents: Zhao, C., H.Wang, Y.Lu, etal. 2019. “Deep Sequencing Reveals Early Reprogramming of Arabidopsis Root Transcriptomes Upon Ralstonia solanacearum Infection.” Molecular Plant Microbe Interactions32: 813 827. Rect: 438, 429, 455, 416 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "figure 2" for your more reference Rect: 414, 395, 418, 382 ActionDest: 146 Annotation: Subtype: Link Contents: Please see "figure 3" for your more reference Rect: 540, 383, 544, 370 ActionDest: 162 Annotation: Subtype: Link Contents: Cao, P., J.Chen, R.Wang, etal. 2022. “A Conserved Type III Effector RipB Is Recognized in Tobacco and Contributes to Ralstonia solanacearum Virulence in Susceptible Host Plants.” Biochemical and Biophysical Research Communications631: 18 24. Rect: 328, 280, 346, 267 ActionDest: 284 Annotation: Subtype: Link Contents: Yu, G., L.Zhang, H.Xue, etal. 2024. “Cell WallMediated Root Development Is Targeted by a SoilBorne Bacterial Pathogen to Promote Infection.” Cell Reports43: 114179. Rect: 388, 280, 406, 267 ActionDest: 288 Annotation: Subtype: Link Contents: Vasse, J., P.Frey, and A.Trigalet. 1995. “Microscopic Studies of Intercellular Infection and Protoxylem Invasion of Tomato Roots by Pseudomonassolanacearum.” Molecular Plant Microbe Interactions8: 241 251. Rect: 327, 257, 345, 244 ActionDest: 288 Annotation: Subtype: Link Contents: Yu, G., L.Zhang, H.Xue, etal. 2024. “Cell WallMediated Root Development Is Targeted by a SoilBorne Bacterial Pathogen to Promote Infection.” Cell Reports43: 114179. Rect: 385, 257, 403, 244 ActionDest: 288 Annotation: Subtype: Link Contents: Kong, X., C.Zhang, H.Zheng, etal. 2020. “Antagonistic Interaction Between Auxin and SA Signaling Pathways Regulates Bacterial Infection Through Lateral Root in Arabidopsis.” Cell Reports32: 108060. Rect: 408, 199, 427, 186 ActionDest: 288 Annotation: Subtype: Link Contents: Zhao, C., H.Wang, Y.Lu, etal. 2019. “Deep Sequencing Reveals Early Reprogramming of Arabidopsis Root Transcriptomes Upon Ralstonia solanacearum Infection.” Molecular Plant Microbe Interactions32: 813 827. Rect: 472, 119, 490, 106 ActionDest: 290 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Contents: Please see "figure 1" for your more reference Rect: 342, 107, 346, 94 ActionDest: 129 Annotation: Subtype: Link Contents: Please see "figure 2e" for your more reference Rect: 352, 107, 361, 94 ActionDest: 146 Annotation: Subtype: Link Contents: Please see "figure 3c" for your more reference Rect: 383, 107, 391, 94 ActionDest: 162 Annotation: Subtype: Link Contents: Please see "figure 6" for your more reference Rect: 383, 72, 387, 59 ActionDest: 213 Page: Sequence: 12 Annotations: Annotation: Subtype: Link Contents: Please see "figure 6e" for your more reference Rect: 175, 141, 183, 128 ActionDest: 213 Annotation: Subtype: Link Contents: Please see "figure 7d" for your more reference Rect: 123, 95, 131, 82 ActionDest: 222 Annotation: Subtype: Link Contents: Please see "figure 6" for your more reference Rect: 411, 486, 416, 473 ActionDest: 213 Annotation: Subtype: Link Contents: Please see "figure 7" for your more reference Rect: 512, 463, 517, 450 ActionDest: 222 Annotation: Subtype: Link Contents: BundalovicTorma, C., F.Lonjon, D.Desveaux, and D. S.Guttman. 2022. “Diversity, Evolution, and Function of Pseudomonas syringae Effectoromes.” Annual Review of Phytopathology60: 211 236. Rect: 467, 383, 485, 370 ActionDest: 284 Annotation: Subtype: Link Contents: Chen, H., J.Chen, M.Li, etal. 2017. “A Bacterial Type III Effector Targets the Master Regulator of Salicylic Acid Signaling, NPR1, to Subvert Plant Immunity.” Cell Host & Microbe22: 777 788. Rect: 387, 348, 405, 335 ActionDest: 284 Annotation: Subtype: Link Contents: Cheng, W., K. R.Munkvold, H.Gao, etal. 2011. “Structural Analysis of Pseudomonas syringae AvrPtoB Bound to Host BAK1 Reveals Two Similar KinaseInteracting Domains in a Type III Effector.” Cell Host & Microbe10: 616 626. Rect: 462, 348, 480, 335 ActionDest: 284 Annotation: Subtype: Link Contents: GimenezIbanez, S., D. R.Hann, V.Ntoukakis, E.Petutschnig, V.Lipka, and J. P.Rathjen. 2009. “AvrPtoB Targets the LysM Receptor Kinase CERK1 to Promote Bacterial Virulence on Plants.” Current Biology19: 423 429. Rect: 328, 337, 347, 324 ActionDest: 288 Annotation: Subtype: Link Contents: Gohre, V., T.Spallek, H.Haweker, etal. 2008. “Plant PatternRecognition Receptor FLS2 Is Directed for Degradation by the Bacterial Ubiquitin Ligase AvrPtoB.” Current Biology18: 1824 1832. Rect: 402, 337, 421, 324 ActionDest: 288 Annotation: Subtype: Link Contents: Wang, M. Y., J. B.Chen, R.Wu, etal. 2023. “The Plant Immune Receptor SNC1 Monitors Helper NLRs Targeted by a Bacterial Effector.” Cell Host & Microbe31: 1792 1803. Rect: 474, 337, 492, 324 ActionDest: 288 Annotation: Subtype: Link Contents: Wang, W., N.Liu, C.Gao, L.Rui, and D.Tang. 2019. “The Pseudomonas syringae Effector AvrPtoB Associates With and Ubiquitinates Arabidopsis Exocyst Subunit EXO70B1.” Frontiers in Plant Science10: 1027. Rect: 497, 337, 515, 324 ActionDest: 288 Annotation: Subtype: Link Contents: Yu, G., M.Derkacheva, J. S.Rufian, etal. 2022. “The Arabidopsis E3 Ubiquitin Ligase PUB4 Regulates BIK1 and Is Targeted by a Bacterial TypeIII Effector.” EMBO Journal41: e107257. Rect: 330, 279, 349, 266 ActionDest: 288 Annotation: Subtype: Link Contents: Yu, G., L.Xian, H.Xue, etal. 2020. “A Bacterial Effector Protein Prevents MAPKMediated Phosphorylation of SGT1 to Suppress Plant Immunity.” PLoS Pathogens16: e1008933. Rect: 355, 279, 374, 266 ActionDest: 288 Annotation: Subtype: Link Contents: Yu, G., L.Zhang, H.Xue, etal. 2024. “Cell WallMediated Root Development Is Targeted by a SoilBorne Bacterial Pathogen to Promote Infection.” Cell Reports43: 114179. Rect: 331, 245, 349, 232 ActionDest: 288 Annotation: Subtype: Link Contents: Sun, Z. M., Q.Zhang, Y. X.Feng, etal. 2023. “The Ralstonia solanacearum Type III Effector RipAW Targets the Immune Receptor Complex to Suppress PAMPTriggered Immunity.” International Journal of Molecular Sciences25: 183. Rect: 383, 222, 401, 209 ActionDest: 288 Annotation: Subtype: Link Contents: GomezGomez, L., and T.Boller. 2000. “FLS2: An LRR ReceptorLike Kinase Involved in the Perception of the Bacterial Elicitor Flagellin in Arabidopsis.” Molecular Cell5: 1003 1011. Rect: 488, 164, 507, 151 ActionDest: 288 Annotation: Subtype: Link Contents: Zhang, J., W.Li, T.Xiang, etal. 2010. “ReceptorLike Cytoplasmic Kinases Integrate Signaling From Multiple Plant Immune Receptors and Are Targeted by a Pseudomonas syringae Effector.” Cell Host & Microbe7: 290 301. Rect: 529, 95, 547, 82 ActionDest: 290 Annotation: Subtype: Link Contents: Zhang, Y., S.Xu, P.Ding, etal. 2010. “Control of Salicylic Acid Synthesis and Systemic Acquired Resistance by Two Members of a PlantSpecific Family of Transcription Factors.” Proceedings of the National Academy of Sciences of the United States of America107: 18220 18225. Rect: 378, 84, 396, 71 ActionDest: 290 Annotation: Subtype: Link Contents: Sun, T., Y.Zhang, Y.Li, Q.Zhang, and Y.Ding. 2015. “ChIPSeq Reveals Broad Roles of SARD1 and CBP60g in Regulating Plant Immunity.” Nature Communications6: 10159. Rect: 444, 84, 463, 71 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 4" for your more reference Rect: 543, 61, 547, 48 ActionDest: 176 Page: Sequence: 13 Annotations: Annotation: Subtype: Link Contents: Figure S3: Identification the expressions of CBP60g and SARD1 in Est::RipAW transgenic line 16#. Genes expressions of CBP60g and SARD1 in Est::RipAW transgenic 16# seedlings were measured by RTqPCR at 48hpe. Gene expression was quantified and normalised to AtActin2 using the 2−Ct method. The experiment was performed three with similar results. Statistical analysis was done with oneway ANOVA Sidka's test (**p<0.01, ***p<0.001). Rect: 74, 751, 83, 739 ActionDest: 290 Annotation: Subtype: Link Contents: Please see "table 1" for your more reference Rect: 131, 751, 135, 739 ActionDest: 180 Annotation: Subtype: Link Contents: Please see "figure 5" for your more reference Rect: 77, 728, 82, 716 ActionDest: 194 Annotation: Subtype: Link Contents: Hu, J., X.Barlet, L.Deslandes, etal. 2008. “Transcriptional Responses of Arabidopsis thaliana During Wilt Disease Caused by the SoilBorne Phytopathogenic Bacterium, Ralstonia solanacearum.” PLoS One3: e2589. Rect: 245, 671, 264, 658 ActionDest: 288 Annotation: Subtype: Link Contents: Zhao, C., H.Wang, Y.Lu, etal. 2019. “Deep Sequencing Reveals Early Reprogramming of Arabidopsis Root Transcriptomes Upon Ralstonia solanacearum Infection.” Molecular Plant Microbe Interactions32: 813 827. Rect: 69, 659, 87, 647 ActionDest: 290 Annotation: Subtype: Link Contents: Qi, P., M.Huang, X.Hu, etal. 2022. “A Ralstonia solanacearum Effector Targets TGA Transcription Factors to Subvert Salicylic Acid Signaling.” Plant Cell34: 1666 1683. Rect: 82, 613, 100, 601 ActionDest: 288 Annotation: Subtype: Link Contents: Nakano, M., and T.Mukaihara. 2018. “Ralstonia solanacearum Type III Effector RipAL Targets Chloroplasts and Induces Jasmonic Acid Production to Suppress Salicylic AcidMediated Defense Responses in Plants.” Plant & Cell Physiology59: 2576 2589. Rect: 93, 590, 110, 578 ActionDest: 288 Annotation: Subtype: Link Contents: Sun, Z. M., Q.Zhang, Y. X.Feng, etal. 2023. “The Ralstonia solanacearum Type III Effector RipAW Targets the Immune Receptor Complex to Suppress PAMPTriggered Immunity.” International Journal of Molecular Sciences25: 183. Rect: 66, 544, 84, 532 ActionDest: 288 Annotation: Subtype: Link Contents: Please see "figure 6e" for your more reference Rect: 113, 521, 121, 509 ActionDest: 213 Annotation: Subtype: Link Contents: Wang, L., K.Tsuda, M.Sato, J. D.Cohen, F.Katagiri, and J.Glazebrook. 2009. “Arabidopsis CaM Binding Protein CBP60g Contributes to MAMPInduced SA Accumulation and Is Involved in Disease Resistance Against Pseudomonas syringae.” PLoS Pathogens5: e1000301. Rect: 243, 314, 262, 302 ActionDest: 288 Annotation: Subtype: Link Contents: Wang, L., K.Tsuda, W.Truman, etal. 2011. “CBP60g and SARD1 Play Partially Redundant Critical Roles in Salicylic Acid Signaling.” Plant Journal67: 1029 1041. Rect: 268, 314, 286, 302 ActionDest: 288 Annotation: Subtype: Link Contents: Zhang, Y., S.Xu, P.Ding, etal. 2010. “Control of Salicylic Acid Synthesis and Systemic Acquired Resistance by Two Members of a PlantSpecific Family of Transcription Factors.” Proceedings of the National Academy of Sciences of the United States of America107: 18220 18225. Rect: 111, 303, 128, 290 ActionDest: 290 Annotation: Subtype: Link Contents: Hu, J., X.Barlet, L.Deslandes, etal. 2008. “Transcriptional Responses of Arabidopsis thaliana During Wilt Disease Caused by the SoilBorne Phytopathogenic Bacterium, Ralstonia solanacearum.” PLoS One3: e2589. Rect: 140, 268, 159, 256 ActionDest: 288 Annotation: Subtype: Link Contents: Zhao, C., H.Wang, Y.Lu, etal. 2019. “Deep Sequencing Reveals Early Reprogramming of Arabidopsis Root Transcriptomes Upon Ralstonia solanacearum Infection.” Molecular Plant Microbe Interactions32: 813 827. Rect: 215, 268, 232, 256 ActionDest: 290 Annotation: Subtype: Link Contents: Sun, T., Y.Zhang, Y.Li, Q.Zhang, and Y.Ding. 2015. “ChIPSeq Reveals Broad Roles of SARD1 and CBP60g in Regulating Plant Immunity.” Nature Communications6: 10159. Rect: 86, 199, 104, 187 ActionDest: 288 Annotation: Subtype: Link Contents: TableS3: Primers used in this study. Rect: 467, 487, 476, 474 ActionDest: 290 Annotation: Subtype: Link Contents: Zhou, Z., G.Bi, and J. M.Zhou. 2018. “Luciferase Complementation Assay for Protein Protein Interactions in Plants.” Current Protocols in Plant Biology3: 42 50. Rect: 353, 430, 371, 417 ActionDest: 290 Annotation: Subtype: Link Contents: TableS3: Primers used in this study. Rect: 514, 407, 522, 394 ActionDest: 290 Annotation: Subtype: Link Contents: Zhou, Z., G.Bi, and J. M.Zhou. 2018. “Luciferase Complementation Assay for Protein Protein Interactions in Plants.” Current Protocols in Plant Biology3: 42 50. Rect: 352, 361, 370, 348 ActionDest: 290 Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 479, 338, 497, 325 ActionDest: 288 Annotation: Subtype: Link Contents: Niu, Y., S.Fu, G.Chen, etal. 2022. “Different Epitopes of Ralstonia solanacearum Effector RipAW Are Recognized by Two Nicotiana Species and Trigger Immune Responses.” Molecular Plant Pathology23: 188 203. Rect: 528, 165, 546, 152 ActionDest: 288 Annotation: Subtype: Link Contents: Choi, K. H., J. B.Gaynor, K. G.White, etal. 2005. “A Tn7Based BroadRange Bacterial Cloning and Expression System.” Nature Methods2: 443 448. Rect: 457, 142, 476, 129 ActionDest: 284 Annotation: Subtype: Link Contents: Zhang, Y., Y.Cao, L.Zhang, Y.Hikichi, K.Ohnishi, and J.Li. 2021. “The Tn7Based Genomic Integration is Dependent on an attTn7 Box in the Glms Gene and is SiteSpecific With Monocopy in Ralstonia solanacearum Species Complex.” Molecular Plant Microbe Interactions34: 720 725. Rect: 526, 142, 544, 129 ActionDest: 290 Annotation: Subtype: Link Contents: TableS3: Primers used in this study. Rect: 457, 73, 466, 60 ActionDest: 290 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Contents: TableS3: Primers used in this study. Rect: 189, 717, 197, 704 ActionDest: 290 Annotation: Subtype: Link Contents: Zhang, Y., Y.Cao, L.Zhang, Y.Hikichi, K.Ohnishi, and J.Li. 2021. “The Tn7Based Genomic Integration is Dependent on an attTn7 Box in the Glms Gene and is SiteSpecific With Monocopy in Ralstonia solanacearum Species Complex.” Molecular Plant Microbe Interactions34: 720 725. Rect: 254, 717, 271, 704 ActionDest: 290 Annotation: Subtype: Link Contents: Teixeira, P., N. R.Colaianni, T. F.Law, etal. 2021. “Specific Modulation of the Root Immune System by a Community of Commensal Bacteria.” Proceedings of the National Academy of Sciences of the United States of America118: e2100678118. Rect: 106, 441, 124, 428 ActionDest: 288 Annotation: Subtype: Link Contents: Wang, X., Q.Gong, S.Cheng, etal. 2024. “Cytokinin Plays a Multifaceted Role in Ralstonia solanacearumTriggered Plant Disease Development.” Molecular Plant Pathology25: e70045. Rect: 179, 441, 198, 428 ActionDest: 288 Annotation: Subtype: Link Contents: Liu, J., S.Fu, L.Yang, etal. 2016. “Vacuolar SPXMFS Transporters Are Essential for Phosphate Adaptation in Plants.” Plant Signaling & Behavior11: e1213474. Rect: 234, 303, 251, 290 ActionDest: 288 Annotation: Subtype: Link Contents: Lu, H., A. S.Lema, M.PlanasMarques, A.AlonsoDiaz, M.Valls, and N. S.Coll. 2018. “Type III SecretionDependent and Independent Phenotypes Caused by Ralstonia solanacearum in Arabidopsis Roots.” Molecular Plant Microbe Interactions31: 175 184. Rect: 66, 268, 83, 256 ActionDest: 288 Annotation: Subtype: Link Contents: Yu, G., L.Xian, H.Xue, etal. 2020. “A Bacterial Effector Protein Prevents MAPKMediated Phosphorylation of SGT1 to Suppress Plant Immunity.” PLoS Pathogens16: e1008933. Rect: 84, 222, 102, 210 ActionDest: 288 Annotation: Subtype: Link Contents: TableS1: Overview of RNAseq data quality. Rect: 365, 705, 374, 693 ActionDest: 290 Annotation: Subtype: Link Contents: Tian, T., Y.Liu, H.Yan, etal. 2017. “agriGO v2.0: A GO Analysis Toolkit for the Agricultural Community, 2017 Update.” Nucleic Acids Research45: W122 W129. Rect: 381, 475, 398, 463 ActionDest: 288 Annotation: Subtype: Link Contents: TableS3: Primers used in this study. Rect: 531, 280, 540, 267 ActionDest: 290 Page: Sequence: 15 Annotations: Annotation: Subtype: Link Contents: Zhou, Z., G.Bi, and J. M.Zhou. 2018. “Luciferase Complementation Assay for Protein Protein Interactions in Plants.” Current Protocols in Plant Biology3: 42 50. Rect: 66, 498, 84, 486 ActionDest: 290 Annotation: Subtype: Link Contents: Chen, H., Y.Zou, Y.Shang, etal. 2008. “Firefly Luciferase Complementation Imaging Assay for ProteinProtein Interactions in Plants.” Plant Physiology146: 368 376. Rect: 131, 498, 150, 486 ActionDest: 284 Page: Sequence: 16 Page: Sequence: 17 Checksum: bdba4bb7 Type: CRC32 Checksum: cc72e83733cea5ddbbf65baae5b17aaa Type: MD5 Checksum: fefb1100dfdc90f658be3f507101ee4e0600a95d Type: SHA-1 Checksum: 620dd00713edaf03d9d9ed9758ce8967fc234f0f6142d76f981e22b0e00396c4 Type: SHA-256