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Fig. 3 | Plant Methods

Fig. 3

From: A cell death assay in barley and wheat protoplasts for identification and validation of matching pathogen AVR effector and plant NLR immune receptors

Fig. 3

Results of example setup transfection (Tables 2 and 3) into wheat (a) and barley (b) mesophyll leaf protoplasts based on four biological replicates. Isolated protoplasts were transfected with pUBQ:luciferase and either a pIPKb002 empty vector (EV) control or pIPKb002 vector with cDNAs of AVRa1, AVRa1-V1, AvrSr50WT, AvrSr50RKQQC, AvrSr50QCMJC all lacking respective signal peptides together with either Mla1 or Sr50. Luciferase activity was determined 16 h post transfection as proxy for cell death. Differences amongst all transfection samples were assessed by analysis of variance and subsequent Tukey post hoc test of luciferase measurements normalised to the EV sample for each NLR construct (EV = 1). Observed P values were as follows: a P = 1.594e−06, b P = 1.573e−07. Samples marked by different letters differ significantly (P < 0.05) in the Tukey test. Experiment was performed four times independently with different plant material used each day and all values (Tables 2 and 3) obtained in the full biological replicates are indicated in turquoise; square: Experiment 1, cross: Experiment 2, triangle: Experiment 3, dot: Experiment 4

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