Procedure

P. vulgaris was inoculated with Rhizobium tropici CIAT 899 and grown hydroaeroponically under P sufficiency versus P deficiency as described by Vadez et al. (1999). Nodules were harvested during vegetative stage, and immediately fixed and embedded in methacrylic resin for in situ hybridization (ISH) during four weeks according to Schumpp (1998). Sense and antisense 35S-UTP-labeled RNA were synthesized from a putative aquaporin-1 (Mip-1) cloned in pGEMT (Promega), using SP6 and T7 polymerase (Promega).

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