Fig. 2: PlPeL1 and PlPeL1-like interact with LcPIP1. | Nature Communications

Fig. 2: PlPeL1 and PlPeL1-like interact with LcPIP1.

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

Fig. 2

a Yeast two-hybrid (Y2H) assays demonstrated that LcPIP1 interacted with PlPeL1 and PlPeL1-like. PlPeL1 and PlPeL1-like were cloned into pGBKT7, and LcPIP1 was cloned into pGADT7 vector. AD, pGADT7 vector; BD, pGBKT7 vector. A combination of BD-53 and AD-T was used as a positive control, and a combination of BD-Lam and AD-T was used as a negative control. Yeast transformants were grown on SD/-Trp/-Leu or SD/-Trp/-Leu/-His/-Ade selective medium with X-a-gal. Images were taken 3 days post-inoculation. b LcPIP1 interacted with PlPeL1 and PlPeL1-like in planta. PlPeL1-HA and PlPeL1-like-HA were co-expressed with LcPIP1-RFP, NbPIP1-RFP, or RFP in N. benthamiana leaves. Protein complexes were immunoprecipitated with RFP-Trap-M beads. Co-precipitation was detected by western blot. c LcPIP1 physically interacted with PlPeL1 and PlPeL1-like in vitro. GST-PlPeL1-, GST-PlPeL1-like-, or GST- bound beads were incubated with bacteria lysate containing His-LcPIP1 or His-NbPIP1. Co-precipitation was detected by western blot. Red asterisks indicated protein bands of the correct size. Ponceau S staining of Rubisco was used to indicate loading quantity of protein samples. All experiments were repeated three times with similar results.

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