Fig. 5: LcPIP1 homologs are widely present in land plants and the VDMASG motif is critical for cell death-inducing activity. | Nature Communications

Fig. 5: LcPIP1 homologs are widely present in land plants and the VDMASG motif is critical for cell death-inducing activity.

From: A plant cell death-inducing protein from litchi interacts with Peronophythora litchii pectate lyase and enhances plant resistance

Fig. 5

a Phylogenetic analysis illustrates the relationship among LcPIP1, its closest paralogous protein (LcPIP1PP), and 23 homologs from diverse species. The tree was constructed using the neighbor-joining algorithm. The proteins that induced cell death in N. benthamiana are indicated by red triangles. b Cell death assays of LcPIP1 homologs expressed in N. benthamiana leaves. Representative N. benthamiana leaves were photographed at 4 dpa. c Western blot analysis of total tissue extracts (TE) and apoplastic fluid (AF) from N. benthamiana leaves agroinfiltrated with LcPIP1-HA or LcPIP1ΔSP-HA for 36 hpa. Extracted proteins were analyzed using immunoblotting with anti-HA or anti-Actin antibodies, as indicated. Red asterisks indicated protein bands of the correct size. Ponceau S staining of Rubisco was used to indicate loading quantity of protein samples. d LcPIP1ΔSP was unable to induce cell death, but the SP of PR1 restored the ability of the LcPIP1 deletion mutant to trigger cell death. Photographs were taken at 4 dpa. e Schematic diagrams of the protein structures of six truncated and site-directed mutants of LcPIP1 were shown on the left. Cell death symptoms and accumulation of ROS in N. benthamiana leaves expressing full-length and mutants were shown on the right. Photographs were taken at 4 dpa. Accumulation of ROS was observed in protein-expressing leaves at 2 dpa. These experiments were repeated three times with similar results.

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