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. 2014 Dec 1;9(12):e113875. doi: 10.1371/journal.pone.0113875

Figure 2. AvrRxo1 induces cell death in N. benthamiana and affects cell growth in yeast.

Figure 2

(A) Yeast growth is inhibited by expression of AvrRxo1. GS115 yeast strains carrying empty vecter pPIC3.5 or pPIC3.5:avrRxo1RS105 were grown overnight in repressing broth (2% glucose) at 30°C. Cultures were then diluted to OD600 = 1.0, and then serial 10-fold dilutions were spotted onto resuppressing or inducing medium (1% methanol). Photographs were taken after 2 days of growth. (B) Phenotype of transient expression of avrRxo1 from different Xoc strains in N. benthamiana (4–6 weeks old). Transient expression of avrRxo1 SDAU-1 (1), avrRxo1 RS105 (2) and avrRxo1 RS85 (5) induces N. benthamiana cell death. No cell death for avrRxo1 JSB2-24 (3) and avrRxo1 HNB8-47 (4) as control pGR106 (6) treatment. (C) The serine residue at the 344 position is required for toxicity of avrRxo1 in N. benthamiana. The transient expression of avrRxo1 from RS105 (1) induced cell death, whereas avrRxo1 from JSB2–24 (2) did not induce tobacco cell death. AvrRxo1(S344T) (3), whose serine residue at the 344 position was substituted to threonine, failed to induce cell death when transiently expressed in N. benthamiana. (D) The normal expression of avrRxo1 in infected leaves were confirmed by RT-PCR. Housekeeping gene EF1a was selected to normalize the samples. Data shown are representative of three independent experiments with similar results.