Extended Data Fig. 1: Expression and traits of transgenic plants of HA-PCSPVY-aTm-22.
From: Remodelling autoactive NLRs for broad-spectrum immunity in plants
a, Transient expression of either NIaPVY-Myc or NIa-ProTuMV-Myc caused cell death in T0 transgenic HA-PCSPVY-aTm-22 plants (lines 3 and 5), but not in wild-type (WT) plants at 2 days post agroinfiltration. Bar is 2 cm. b, Successful expression of Myc-tagged proteins was confirmed by immunoblotting. Loading of total protein samples for analyses are shown (lower panel). Sizes and positions of protein markers are indicated. c, T1 transgenic HA-PCSPVY-aTm-22 plants of two independent lines showed no developmental defects. Bar is 5 cm. d, No significant trait difference between wild-type and transgenic plants. Height and fresh weight were measured when the plants are 3 months old, and seed setting indicated by numbers of seed pods were measured when the plants are 4 months old. “ns” means no significance as determined by two-sided Student’s t tests (n = 6 biologically independent samples). Data are represented as mean ± s.d. e, Transgene and protein expression of the HA-PCSPVY-aTm-22 chimeric protein were detected by genomic PCR (upper panel) and immunoblotting (middle panel) in three independent individuals of T1 plants from two independent transgenic lines 3 and 5. Loading of total protein samples for analyses are shown (lower panel). Sizes and positions of DNA (upper panel) and protein markers (middle and lower panels) are indicated. f, Cleavage of HA-PCSPVY-aTm-22 by Myc-NIa-ProPVY in T1 transgenic plants. Myc-NIa-ProPVY and AvrRpt2-Myc were expressed at the left or right halves of the same leaves from the transgenic plants. Wide-type leaf tissues without agroinfiltration served as negative controls. Total proteins were extracted and analyzed. NLR proteins and proteases were detected by anti-HA and anti-Myc antibodies, respectively. Experiments were repeated at least three times with similar results.
