Table S3.
Plasmids generated in this study
| Use | Construct | Plasmid name | Insert or PCR product | Primers | Template | Plasmid backbone | Cloning method | Source |
| Entry clones for N. benthamiana assays | MLA101–160 | pSC260 | MLA101–160 (without stop) | oCS195/197 | Plasmid (cDNA) from Paul Schulze-Lefert* (13) | pDONR207 | BP Gateway (Life Technologies) | – |
| MLA101–120 | pSC323 | MLA101–120 (without stop) | oCS195/211 | Plasmid (cDNA) from Paul Schulze-Lefert (13) | pDONR207 | BP Gateway | – | |
| MLA101–225 | pSC324 | MLA101–225 (without stop) | oCS195/214 | Plasmid (cDNA) from Paul Schulze-Lefert (13) | pDONR207 | BP Gateway | – | |
| Sr331–160 | pSC298 | Sr331–160 (without stop) | oCS204/206 | Plasmid (cDNA) from S.P. (10) | pDONR207 | BP Gateway | – | |
| Sr331–120 | pSC325 | Sr331–120 (without stop) | oCS204/212 | Plasmid (cDNA) from S.P. (10) | pDONR207 | BP Gateway | – | |
| Sr331–225 | pSC326 | Sr331–225 (without stop) | oCS204/215 | Plasmid (cDNA) from S.P. (10) | pDONR207 | BP Gateway | – | |
| Sr331–160 A122E/I123E | pSC437 | Sr331–160 A122E/I123E (without stop) | oCS317/318 | pSC298 | pDONR207 | QuikChange Lightning (Agilent Technologies) | – | |
| Sr331–160 I126E | pSC438 | Sr331–160 I126E (without stop) | oCS320/321 | pSC298 | pDONR207 | QuikChange Lightning | – | |
| Sr331–160 L130E | pSC439 | Sr331–160 L130E (without stop) | oCS322/323 | pSC298 | pDONR207 | QuikChange Lightning | – | |
| Sr331–160 V133E/A134E | pSC440 | Sr331–160 V133E/A134E (without stop) | oCS324/325 | pSC298 | pDONR207 | QuikChange Lightning | – | |
| Sr331–160 RRDRNK/AAAAAAA | pSC441 | Sr331–160 RRDRNK/AAAAAAA (without stop) | oCS326/327 | pSC298 | pDONR207 | QuikChange Lightning | – | |
| Sr331–160 F142E | pSC442 | Sr331–160 F142E (without stop) | oCS328/329 | pSC298 | pDONR207 | QuikChange Lightning | – | |
| Sr33 full-length | Sr33 full length entry (C.C.) | Sr33 full-length (without stop) | Sr33 attB1-F/Sr33 attB2-R | pGBCT-Sr33 (cDNA) from S.P. (10) | pDONR207 | BP Gateway | – | |
| Sr33 full-length A122E/I123E | pSC443 | Sr33 full-length A122E/I123E (without stop) | oCS317/318 | C.C. Sr33 full-length entry | pDONR207 | QuikChange Lightning | – | |
| Sr33 full-length I126E | pSC444 | Sr33 full-length I126E (without stop) | oCS320/321 | C.C. Sr33 full-length entry | pDONR207 | QuikChange Lightning | – | |
| Sr33 full-length L130E | pSC445 | Sr33 full-length L130E (without stop) | oCS322/323 | C.C. Sr33 full-length entry | pDONR207 | QuikChange Lightning | – | |
| Sr33 full-length V133E/A134E | pSC446 | Sr33 full-length V133E/A134E (without stop) | oCS324/325 | C.C. Sr33 full-length entry | pDONR207 | QuikChange Lightning | – | |
| Sr33 full-length RRDRNK/AAAAAAA | pSC447 | Sr33 full-length RRDRNK/AAAAAAA (without stop) | oCS326/327 | C.C. Sr33 full-length entry | pDONR207 | QuikChange Lightning | – | |
| Sr33 full-length F142E | pSC448 | Sr33 full-length F142E (without stop) | oCS328/329 | C.C. Sr33 full-length entry | pDONR207 | QuikChange Lightning | – | |
| Sr501–163 | pSC262 | Sr501–163 (without stop) | oCS184/187 | Plasmid (Sr50 genomic) from R.M. (11) | pDONR207 | BP Gateway | – | |
| Sr501–123 | pSC327 | Sr501–123 (without stop) | oCS184/213 | Plasmid (Sr50 genomic) from R.M. (11) | pDONR207 | BP Gateway | – | |
| Sr501–228 | pSC328 | Sr501–228 (without stop) | oCS184/216 | Plasmid (Sr50 genomic) from R.M. (11) | pDONR207 | BP Gateway | – | |
| Sr50 full length | Sr50 full length entry (C.C.) | Sr50 full length (without stop) | Sr50 attB1-F/Sr50 attB2-R | pGEMT-Sr50 (cDNA) from R.M. (11) | pDONR207 | BP Gateway | – | |
| Cell-death assays, co-IPs, and confocal microscopy in N. benthamiana | RGA4:HA | pSC061 | RGA4 (cDNA) | – | – | – | – | (46) |
| RGA4:CFP | pSC081 | RGA4 (cDNA) | – | – | – | – | (46) | |
| RGA4MHD:HA | pSC285 | RGA4MHD | – | – | – | – | (46) | |
| RGA4MHD:CFP | pSC306 | RGA4MHD | – | – | – | – | (46) | |
| RGA41–171:CFP | pSC167 | RGA41–171 | – | – | – | – | (46) | |
| MLA101–160:HA | pSC278 | MLA101–160 | – | pSC260 | pBin19-35s::GTW:HA | LR Gateway (Life Technologies) | – | |
| MLA101–160:CFP | pSC302 | MLA101–160 | – | pSC260 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| MLA101–160:YFP:NLS | pSC311 | MLA101–160 | – | pSC260 | pAM-PAT-35s::GTW:YFP:NLS | LR Gateway | – | |
| MLA101–160:YFP:nls | pSC313 | MLA101–160 | – | pSC260 | pAM-PAT-35s::GTW:YFP:nls | LR Gateway | – | |
| MLA101–160:YFP:NES | pSC312 | MLA101–160 | – | pSC260 | pXCSG-35s::GTW:YFP:NES (54) | LR Gateway | – | |
| MLA101–160:YFP:nes | pSC314 | MLA101–160 | – | pSC260 | pXCSG-35s::GTW:YFP:nes (54) | LR Gateway | – | |
| MLA101–225:HA | pSC338 | MLA101–225 | – | pSC324 | pBin19-35s::GTW:HA | LR Gateway | – | |
| MLA101–120:HA | pSC337 | MLA101–120 | – | pSC323 | pBin19-35s::GTW:HA | LR Gateway | – | |
| MLA101–120:CFP | pSC343 | MLA101–120 | – | pSC323 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160:HA | pSC300 | Sr331–160 | – | pSC298 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160:CFP | pSC301 | Sr331–160 | – | pSC298 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160:YFP:NLS | pSC315 | Sr331–160 | – | pSC298 | pAM-PAT-35s::GTW:YFP:NLS | LR Gateway | – | |
| Sr331–160:YFP:nls | pSC317 | Sr331–160 | – | pSC298 | pAM-PAT-35s::GTW:YFP:nls | LR Gateway | – | |
| Sr331–160:YFP:NES | pSC316 | Sr331–160 | – | pSC298 | pXCSG-35s::GTW:YFP:NES (54) | LR Gateway | – | |
| Sr331–160:YFP:nes | pSC318 | Sr331–160 | – | pSC298 | pXCSG-35s::GTW:YFP:nes (54) | LR Gateway | – | |
| Sr331–120:HA | pSC339 | Sr331–120 | – | pSC325 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–120:CFP | pSC345 | Sr331–120 | – | pSC325 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–225:HA | pSC340 | Sr331–225 | – | pSC326 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 A122E/I123E:HA | pSC449 | Sr331–160 A122E/I123E | – | pSC437 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 I126E:HA | pSC450 | Sr331–160 I126E | – | pSC438 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 L130E:HA | pSC451 | Sr331–160 L130E | – | pSC439 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 V133E/A134E:HA | pSC452 | Sr331–160 V133E/A134E | – | pSC440 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 R136A/R137A/D138A/R139A/N140A/K141A:HA | pSC453 | Sr331–160 R136A/R137A/D138A/R139A/N140A/K141A | – | pSC441 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 F142E:HA | pSC454 | Sr331–160 F142E | – | pSC442 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr331–160 A122E/I123E:CFP | pSC455 | Sr331–160 A122E/I123E | – | pSC437 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160 I126E:CFP | pSC456 | Sr331–160 I126E | – | pSC438 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160 L130E:CFP | pSC457 | Sr331–160 L130E | – | pSC439 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160 V133E/A134E:CFP | pSC458 | Sr331–160 V133E/A134E | – | pSC440 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160 RRDRNK/AAAAAAA:CFP | pSC459 | Sr331–160 RRDRNK/AAAAAAA | – | pSC441 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr331–160 F142E:CFP | pSC460 | Sr331–160 F142E | – | pSC442 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr33 full-length:HA | Sr33:HA (C.C.) | Sr33 full-length | – | C.C. Sr33 full-length entry | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr33 full length:CFP | Sr33:CFP (C.C.) | Sr33 full length | – | C.C. Sr33 full length entry | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr33 full-length A122E/I123E:HA | pSC461 | Sr33 full-length A122E/I123E | – | pSC443 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr33 full-length I126E:HA | pSC462 | Sr33 full-length I126E | – | pSC444 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr33 full-length L130E:HA | pSC463 | Sr33 full-length L130E | – | pSC445 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr33 full-length V133E/A134E:HA | pSC464 | Sr33 full-length V133E/A134E | – | pSC446 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr33 full-length RRDRNK/AAAAAAA:HA | pSC465 | Sr33 full-length RRDRNK/AAAAAAA | – | pSC447 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr33 full-length F142E:HA | pSC466 | Sr33 full-length F142E | – | pSC448 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr501–163:HA | pSC280 | Sr501–163 | – | pSC262 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr501–163:CFP | pSC303 | Sr501–163 | – | pSC262 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr501–163:YFP:NLS | pSC319 | Sr501–163 | – | pSC262 | pAM-PAT-35s::GTW:YFP:NLS | LR Gateway | – | |
| Sr501–163:YFP:nls | pSC321 | Sr501–163 | – | pSC262 | pAM-PAT-35s::GTW:YFP:nls | LR Gateway | – | |
| Sr501–163:YFP:NES | pSC320 | Sr501–163 | – | pSC262 | pXCSG-35s::GTW:YFP:NES (54) | LR Gateway | – | |
| Sr501–163:YFP:nes | pSC322 | Sr501–163 | – | pSC262 | pXCSG-35s::GTW:YFP:nes (54) | LR Gateway | – | |
| Sr501–123:HA | pSC341 | Sr501–123 | – | pSC327 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr501–123:CFP | pSC347 | Sr501–123 | – | pSC327 | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Sr501–228:HA | pSC342 | Sr501–228 | – | pSC328 | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr50 full-length:HA | Sr50:HA (C.C.) | Sr50 full-length | – | C.C. Sr50 full-length entry | pBin19-35s::GTW:HA | LR Gateway | – | |
| Sr50 full-length:CFP | Sr50:CFP (C.C.) | Sr50 full-length | – | C.C. Sr50 full-length entry | pBin19-35s::GTW:CFP | LR Gateway | – | |
| Yeast two-hybrid assay | GAL4BD:cMyc:Sr501–163 | BD:Sr501–163 (J.M.) | Sr501–163 | Sr50F EcoRI/ Sr50163R Xhol | Plasmid (Sr50 cDNA) from R.M. | pGBKT7 | Direct cloning (EcoRI/Xbal) | – |
| GAL4AD:HA:Sr501–163 | AD:Sr501–163 (J.M.) | Sr501–163 | Sr50F EcoRI/ Sr50163R Xhol | Plasmid (Sr50 cDNA) from R.M. | pGADT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4BD:cMyc:Sr501–228 | BD:Sr501–228 (J.M.) | Sr501–228 | Sr50F EcoRI/ Sr50228R Xhol | Plasmid (Sr50 cDNA) from R.M. | pGBKT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4AD:HA:Sr501–228 | AD:Sr501–228 (J.M.) | Sr501–228 | Sr50F EcoRI/ Sr50228R Xhol | Plasmid (Sr50 cDNA) from R.M. | pGADT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4BD:cMyc:MLA101–160 | BD:MLA101–160 (J.M.) | MLA101–160 | MLA10F EcoRI/ MLA10160R Xhol | Plasmid (cDNA) from Paul Schulze-Lefert (13) | pGBKT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4AD:HA:MLA101–160 | AD:MLA101–160 (J.M.) | MLA101–160 | MLA10F EcoRI/ MLA10160R Xhol | Plasmid (cDNA) from Paul Schulze-Lefert (13) | pGADT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4BD:cMyc:MLA101–225 | BD:MLA101–225 (J.M.) | MLA101–225 | MLA10F EcoRI/ MLA10225R Xhol | Plasmid (cDNA) from Paul Schulze-Lefert (13) | pGBKT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4AD:HA:MLA101–225 | AD:MLA101–225 (J.M.) | MLA101–225 | MLA10F EcoRI/ MLA10225R Xhol | Plasmid (cDNA) from Paul Schulze-Lefert 13) | pGADT7 | Direct cloning (EcoRI/Xbal) | – | |
| GAL4BD:cMyc:RGA51–228 | BD:RGA5 (M.B.) | RGA51–228 | – | – | – | – | (16) | |
| GAL4AD:HA:RGA51–228 | AD:RGA5 (M.B.) | RGA51–228 | – | – | – | – | (16) | |
| GAL4BD:cMyc:L6_TIR | BD:L6-TIR (M.B.) | L6-TIR | – | – | – | – | (7) | |
| GAL4AD:HA:L6_TIR | AD:L6-TIR (M.B.) | L6-TIR | – | – | – | – | (7) | |
| Binary GWY vectors construction for N. benthamiana assays | – | pAM-PAT-35s::GTW:YFPv:NLS | Sma1-YFP-NLS-Xba1 | SmaI ccdB Fwd/ Xba1 NLS-YFP rev | pAM-PAT-35s::GTW:YFPv | pAM-PAT-35s::GTW:YFPv (53) | Direct cloning (Sma1/Xbal) | – |
| – | pAM-PAT-35s::GTW:YFPv:nLs | Sma1-YFP-nls-Xba1 | SmaI ccdB Fwd/ Xba1 nls-YFP rev | pAM-PAT-35s::GTW:YFPv | pAM-PAT-35s::GTW:YFPv (53) | Direct cloning (Sma1/Xbal) | – | |
| Stable wheat transformation and/or cell-death assays in N. benthamiana | pSr33::Sr33:YFPv:NLS:terSr33 | PC17 (J.M.) | Not1-pSr33::Sr33(without stop)-Apa1 | Sr33 Prom NotI-F/ Sr33 Full-length ApaI-R | plasmid (genomic DNA) from S.P. (10) | pVecNeo | Some fragments were ligated in pGEM-T easy vector (Promega) and sequentially ligated in pVecNeo (Apa1/BamH1 and Not1/Apa1) | – |
| Apa1-YFPv:NLS(with stop)-EcoR1 | Venus ApaI Link-F/ Venus-w-NLS-R EcoRI | pAM-PAT-35s::GTW:YFPv | – | |||||
| EcoR1-Sr33ter-BamH1 | Sr33 Term EcoRI-F/ Sr33 Term BamHI-R | Plasmid (genomic DNA) from S.P. (10) | – | |||||
| pSr33::Sr33:YFPv:nls:terSr33 | PC19 (J.M.) | Not1-pSr33::Sr33(without stop)-Apa1 | Sr33 Prom NotI-F/ Sr33 Full-length ApaI-R | Plasmid (genomic DNA) from S.P. (10) | pVecNeo | Some fragments were ligated in pGEM-T easy vector (Promega) and sequentially ligated in pVecNeo (Apa1/BamH1 and Not1/Apa1) | – | |
| Apa1-YFPv:nls(with stop)-EcoR1 | Venus ApaI Link-F/ Venus-m-nls-R EcoRI | pAM-PAT-35s::GTW:YFPv | – | |||||
| EcoR1-Sr33ter-BamH1 | Sr33 Term EcoRI-F/ Sr33 Term BamHI-R | Plasmid (genomic DNA) from S.P. (10) | – | |||||
| pSr33::Sr33:YFPv:NES:terSr33 | PC24 (J.M.) | Not1-pSr33::Sr33(without stop)-Apa1 | Sr33 Prom NotI-F/ Sr33 Full-length ApaI-R | Plasmid (genomic DNA) from S.P. (10) | pVecNeo | Some fragments were ligated in pGEM-T easy vector (Promega) and sequentially ligated in pVecNeo (Apa1/BamH1 and Not1/Apa1) | – | |
| Apa1-YFPv:NES(with stop)-EcoR1 | Venus ApaI Link-F/ Venus-NES2- EcoRI | pAM-PAT-35s::GTW:YFPv | – | |||||
| EcoR1-Sr33ter-BamH1 | Sr33 Term EcoRI-F/ Sr33 Term BamHI-R | Plasmid (genomic DNA) from S.P. (10) | – | |||||
| pSr33::Sr33:YFPv:nes:terSr33 | PC26 (J.M.) | Not1-pSr33::Sr33(without stop)-Apa1 | Sr33 Prom NotI-F/ Sr33 Full-length ApaI-R | Plasmid (genomic DNA) from S.P. (10) | pVecNeo | Some fragments were ligated in pGEM-T easy vector Promega) and sequentially ligated in pVecNeo (Apa1/BamH1 and Not1/Apa1) | – | |
| Apa1-YFPv:nes(with stop)-EcoR1 | Venus ApaI Link-F/ Venus-m-nesR EcoRI | pAM-PAT-35s::GTW:YFPv | – | |||||
| EcoR1-Sr33ter-BamH1 | Sr33 Term EcoRI-F/ Sr33 Term BamHI-R | Plasmid (genomic DNA) from S.P. (10) | – | |||||
| pSr33::Sr33:YFPv:terSr33 | PC16 (J.M.) | Not1-pSr33::Sr33(without stop)-Apa1 | Sr33 Prom NotI-F/Sr33 full-length ApaI-R | Plasmid (genomic DNA) from S.P. (10) | pVecNeo | Some fragments were ligated in pGEM-T easy vector (Promega) and sequentially ligated in pVecNeo (Apa1/BamH1 and Not1/Apa1) | – | |
| Apa1-YFPv(with stop)-EcoR1 | Venus ApaI Link-F/Venus-full-length-R EcoRI | pAM-PAT-35s::GTW:YFPv | – | |||||
| EcoR1-Sr33ter-BamH1 | Sr33 Term EcoRI-F/ Sr33 Term BamHI-R | Plasmid (genomic DNA) from S.P. (10) | – | |||||
| pSr33::YFPv:terSr33 | PC25 (J.M.) | Not1-pSr33(without stop)-Apa1 | Sr33 Prom NotI-F/Sr33 PromR-ApaI | Plasmid (genomic DNA) from S.P. (10) | pVecNeo | Some fragments were ligated in pGEM-T easy vector (Promega) and sequentially ligated in pVecNeo (Apa1/BamH1 and Not1/Apa1) | – | |
| Apa1-YFPv(with stop)-EcoR1 | Venus ApaI Link-F/Venus-full-length-R EcoRI | pAM-PAT-35s::GTW:YFPv | ||||||
| EcoR1-Sr33ter-BamH1 | Sr33 Term EcoRI-F/ Sr33 Term BamHI-R | Plasmid (genomic DNA) from S.P. (10) | ||||||
| pSr50::Sr50g:terSr50 | Sr50g | – | – | – | – | – | (11) | |
| pSr50::Sr50:YFPv:terSr50 | PC137 (C.C.) | PCR 1: Sr50 fragment including internal BaeI site and Sr50 c-ter (without stop)-XmaI-ApaI | Sr50F1/Sr50R1 | Plasmid (genomic DNA) from R.M. (11) | pVecNeo | pGEM-T easy with Sr50 genomic sequence (11) was digested with BaeI and ApaI and ligated to PCR 1. PCR 2 was added using XmaI/ApaI sites then PCR3 was added using XmaI/PmeI sites. The NotIpSr50::Sr50g:YFPv:terSr50-ApaI fragment was ligated into pVecNeo | – | |
| PCR 2: XmaI-PmeI-Sr50ter-ApaI | Sr50F2/Sr50R2 | Plasmid (genomic DNA) from R.M. (11) | ||||||
| PCR 3: XmaI-YFPv(with stop)-PmeI | VenusF/VenusR | pAM-PAT-35s::GTW:YFPv | ||||||
| pSr33::Sr33g:terSr33 | Sr33g | – | – | – | – | – | (10) |
*
Max-Planck Institute, Cologne.