Figure 4: Analysis of the arlpk1-1 arlpk2-1 double mutant.
(a) Phenotype of 4-week-old arlpk1-1 arlpk2-1 double mutants compared with Col-0 WT plants. (b) Bacterial growth in Pst (EV)- and Pst (avrRpt2)-infected Col-0 WT and arlpk1-1 arlpk2-1 double mutants. Bacterial inoculum concentration: 5 × 105 c.f.u. per ml. Bacterial growth was measured at 3 d.p.i. Error bars represent s.d. for at least 15 leaf discs. Similar results were obtained in three biological replicates. **P value <0.01 (Student’s t-test). (c) PR1 protein levels were detected in Pst (EV)- and Pst (avrRpt2)-infected Col-0 WT and arlpk1-1 arlpk2-1 double mutants by western blot using an anti-PR1 antibody. Bacterial inoculum concentration: 1 × 107 c.f.u. per ml. Samples were collected at 0 and 12 h.p.i. α-Tubulin was used as a loading control. Similar results were obtained from two biological replicates. (d) Levels of ARLPK1 transcript in Col-0 WT, mARLPK1-OE1 and mARLPK1-OE2 lines were detected by real-time RT–PCR. Error bars indicate s.d. from three technical replicates. Similar results were obtained in three biological replicates. **P value <0.01 (Student’s t-test). (e) Bacterial growth in Pst (EV)- and Pst (avrRpt2)-infected Col-0 WT and mARLPK1-OE plants. Bacterial inoculum concentration: 5 × 105 c.f.u. per ml. Bacterial growth was measured 3 d.p.i. Error bars represent s.d. for at least 15 leaf discs. Similar results were obtained from three biological replicates. **P value <0.01 (Student’s t-test). (f) Recombinant MBP-tagged kinase domains of RIPK, ARLPK1 and ARLPK2 proteins were subjected to a radioactive kinase assay and kinase activity was detected using autoradiography. Bottom panel: SDS–PAGE gel stained with coomassie brilliant blue (CBB) demonstrating protein abundance. Similar results were obtained from two biological replicates.
