Native Endophytic Pseudomonas putida as a Biocontrol Agent against Common Bean Rust Caused by Uromyces appendiculatus

Abstract

This study aimed to evaluate the efficacy of endophytic bacterium to control common bean rust disease under greenhouse conditions. Endophytic bacterium <i>Pseudomonas putida</i> ASU15 was isolated from fresh asymptomatic common bean, identified using biochemical and molecular characteristics. In vitro, the inhibitory effect of different concentrations of <i>P. putida</i> (1 × 10⁴, 1 × 10⁵ and 1 × 10⁶), as well as fungicide ortiva (0.01%) on uredospores germination of <i>Uromyces appendiculatus</i> were tested using water agar medium. The concentration showing the highest reduction of uredospores germination was at 1 × 10⁶, while there was complete inhibition of uredospores germination associated with using ortiva. Scanning electron microscope exhibited the ability of <i>P. putida</i> cells to attack the cell wall of the fungal uredospores germ tubes of <i>U. appendiculatus</i>, causing obvious cell wall breakdown. The activities of chitinase, lipase, and protease produced by <i>P. putida</i> ASU15, in vitro, were evaluated spectrophotometrically. Chitinolytic, proteolytic, and lipolytic activities were exhibited, contributing 55.26, 3.87, and 26.12 U/mL, respectively. Under greenhouse conditions, treated plants with <i>P. putida</i> ASU15 (two days before pathogen inoculation or at the same time of pathogen inoculation) or fungicide reduced the disease severity, compared to the control. Applying <i>P. putida</i> ASU15 at the same time of pathogen inoculation showed reduction in disease severity (69.9%), higher than application before pathogen inoculation (54.9%). This study is considered the first report that demonstrates the mycoparasitic strategy of <i>P. putida</i> for controlling <i>U. appendiculatus</i>. In conclusion, our results revealed that <i>P. putida</i> ASU15 affords a significant disease reduction that may be attributed to direct suppression of pathogen spores germination.