Systemic Colonization and Expression of Disease Symptoms on Bittersweet Nightshade (Solanum dulcamara) Infected with a GFP-Tagged Dickeya solani IPO2222 (IPO2254)

Abstract

Colonization of Solanum dulcamara (bittersweet nightshade) plants by a GFP-tagged Dickeya solani type strain IPO2222 (IPO2254) was investigated by selective plating and epifluorescence stereomicroscopy (ESM), using in vitro plants and plants grown in compost soil. Replicated experiments were carried out in a growth chamber and the progress of infection and disease symptoms on tissue of the cultured plants, following leaf- and stem-base inoculations with bacteria, was evaluated. Microscopy observations were confirmed by spread-plating dilutions of plant extracts onto agar medium directly after the harvest. In experiments where the stem base of in vitro plants inoculated with a range of inocula of D. solani (104 to 108 colony forming units [cfu] ml-1) was examined at 14 days post infection (dpi), blackleg-like symptoms developed in more than 80% plants together with a reduction of the plant fitness (disease symptoms, weight, height, and appearance). In leaf-inoculated plants at 14 dpi, 15% of the plants exhibited severe blackleg-like symptoms. In detached S. dulcamara leaf assays, IPO2254 survived on the adaxial surface for 14 days at populations of 106 cfu per leaf. Thirty days after stem inoculation of plants grown in compost soil in pots, up to 104 cfu g-1 of GFP-tagged D. solani were found inside the stems. D. solani were detected inside the vascular tissue (xylem vessels) of stems, in the pith tissue in roots, and on the internal surface of the stem hollow. The implications of S. dulcamara infection by D. solani for the long-distance dispersal of the bacterial inoculum are discussed.