Title：
The pH of the leaf apoplast is critical for the formation of Pseudomonas syringae-induced lesions on leaves of the common bean (Phaseolus vulgaris)
Inoculating a virulent strain of Pseudomonas syringae pv. phaseolicola (Pph) into the leaf of common bean (Phaseolus vulgaris) causes the leaf apoplast to alkalinize. Whether or not this apoplastic pH event facilitates virulence of Pph in interaction with common bean is unclear. For elucidating this topic, (i) Pph colonization of the common bean leaf apoplast, (ii) the formation of bacterial lesions, and (iii) apoplastic sucrose concentration were investigated in relation to the apoplastic leaf pH. For this, the Pph-induced leaf apoplastic alkalinization was attenuated by spray application of either a synthetic auxin or an acidic pH buffer. Apoplastic pH was quantified in planta via microscopy-based pH imaging. Apoplastic washing fluids were extracted to quantify both colonization of bacteria in leaf apoplast and the concentration of apoplastic sucrose. Results reveal that the apoplastic alkalinization facilitated bacterial colonization of the apoplast. Number of colony forming units and area of bacterial lesions were reduced when Pph-induced apoplastic alkalinization was attenuated by foliar application of a synthetic auxin or acidic pH buffer. Application of both agents attenuated the Pph-induced increase of sucrose in the leaf apoplast, which is nutrient for bacteria. Data demonstrate that the Pph-mediated leaf apoplastic alkalinisation favours bacterial colonization.
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