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Population Behavior Analysis of dspE and pelD Regulation in Erwinia chrysanthemi 3937

¹Department of Biological Sciences, University of Wisconsin-Milwaukee 53211, U.S.A.; ²Department of Plant Pathology, University of Wisconsin-Madison 53706, U.S.A.; ³Department of Plant Pathology, University of California, Riverside 92521, U.S.A.

Erwinia chrysanthemi 3937 (Ech3937) is a phytopathogenic bacterium with a wide host range. The pectinolytic enzymes secreted by the bacterium and the type III secretion system (T3SS) are essential for full virulence. We used the green fluorescent protein gene as a reporter to investigate the expression of dspE (a putative T3SS effector) and pelD (a major pectin-degrading enzyme) in populations of Ech3937 under different conditions. Gene expression was analyzed by measuring the fluorescence intensity of individual cells with a fluorescence-activated cell sorter. Ech3937 dspE was induced in minimal medium (MM) with only a portion of Ech3937 cells (43.03%) expressing dspE after 12 h of culture. The nutrient-rich King's medium B did not fully eliminate the expression of dspE; a small percentage of Ech3937 cells (5.55%) was able to express dspE after 12 h of culture in this medium. In all, 68.95% of Ech3937 cells expressed pelD after 12 h of culture in MM supplemented with polygalacturonic acid (PGA). However, 96.34% of Ech131 cells (an hrpL deletion mutant of Ech3937) expressed pelD after 12 h of culture in MM supplemented with PGA. In potato tubers, 6.32% of the bacterial cells expressed dspE 2 h after inoculation, whereas only 0.25% of the cells expressed pelD. However, after 24 h, the percentage of cells expressing pelD (68.48%) was approximately 3.5 times that of cells expressing dspE (19.39%). In contrast to potato tubers, similar proportion of Ech3937 cells expressing dspE (39.34%) and pelD (40.30%) were observed in Chinese cabbage 24 h after inoculation. From promoter activity and real-time quantitative results, the expression of pelD in Ech3937 was demonstrated to be downregulated by HrpL in MM supplemented with PGA.

Additional keyword: pectinase.