Fig. 5: Identification of R02860 and R04373 that recognize the RXLR effectors PITG_02860 and PITG_04373. | Nature Genetics

Fig. 5: Identification of R02860 and R04373 that recognize the RXLR effectors PITG_02860 and PITG_04373.

From: Solanum americanum genome-assisted discovery of immune receptors that detect potato late blight pathogen effectors

Fig. 5: Identification of R02860 and R04373 that recognize the RXLR effectors PITG_02860 and PITG_04373.The alt text for this image may have been generated using AI.

a, PITG_02860 is an RXLR effector from P. infestans. An illustration and predicted structure are shown. b, An F2 population was generated from a cross between SP2271 (responds to PITG_02860, R) and SP2272 (no response to PITG_02860, NR). The R bulk (104 progenies) and NR bulk (34 progenies) were used for BSA-RenSeq. c, A total of 218 linked SNPs (red dots) on the top of chromosome 4 of SP2271 were identified. The gray bar represents the chromosome. The physical positions (in Mb) are shown by number. Five molecular markers (S30, S42, S35, S36 and S31) were used for the map-based cloning. The number of recombination events per total tested gametes is shown. d, HR assay of the candidate PITG_02860 receptor. The candidate genes were expressed alone, or co-expressed with PVX::PITG_02860 in N. tabacum leaves. Rpi-amr3 and AVRamr3 were used as controls. NLR16 turned out to be the PITG_02860 receptor (R02860 hereafter). OD600 = 0.5. Four-week-old N. tabacum plants were used, and the photos were taken at 4 dpi. Three biological replicates were performed with the same results. e, PITG_04373 is an RXLR effector from P. infestans. An illustration and predicted structure are shown. f, Both backcross (BC1) and F2 populations were generated from SP2271 and SP2300. The BC1 population of 192 responsive plants and 182 non-responsive progenies were bulked for BSA-RenSeq. The F2 populations were used for fine mapping. g, Informative SNPs (red dots) on the top of chromosome 10 of SP2271 were identified. Five molecular markers (S11, S13, S5, S7 and S16) were used for fine mapping. The number of recombination gametes per total tested gametes is shown. Nine genes from SP2300 SMRT-RenSeq assemblies that mapped to the mapping interval of the SP2271 genome were regarded as candidate genes. All candidates belong to the Rpi-chc1 family except C444. h, HR assay of the PITG_04373 receptor candidates. The candidate genes were expressed alone or co-expressed with 35S::PITG_04373 in N. benthamiana leaves, Rpi-amr3 and AVRamr3 were used as controls. C168 turned out to be the PITG_04373 receptor (R04373 hereafter). OD600 = 0.5. Four-week-old N. benthamiana plants were used and photos were taken at 4 dpi. Three biological replicates were performed with the same results.

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