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

This study aimed to evaluate the efficacy of endophytic bacterium to control common bean rust disease under greenhouse conditions. Endophytic bacterium Pseudomonas putida ASU15 was isolated from fresh asymptomatic common bean, identified using biochemical and molecular characteristics. In vitro, the inhibitory effect of different concentrations of P. putida (1 × 104, 1 × 105 and 1 × 106), as well as fungicide ortiva (0.01%) on uredospores germination of Uromyces appendiculatus 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 P. putida cells to attack the cell wall of the fungal uredospores germ tubes of U. appendiculatus, causing obvious cell wall breakdown. The activities of chitinase, lipase, and protease produced by P. putida 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 P. putida 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 P. putida 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 P. putida for controlling U. appendiculatus. In conclusion, our results revealed that P. putida ASU15 affords a significant disease reduction that may be attributed to direct suppression of pathogen spores germination.

Fungicidal Efficacy of Drying Plant Oils in Green Beans against Bean Rust (Uromyces appendiculatus)

As biorationals, plant oils offer numerous advantages such as being natural products, with low ecotoxicological side effects, and high biodegradability. In particular, drying glyceride plant oils, which are rich in unsaturated fatty acids, might be promising candidates for a more sustainable approach in the discussion about plant protection and the environment. Based on this, we tested the protective and curative efficacy of an oil-in-water-emulsion preparation using drying plant oils (linseed oil, tung oil) and a semi-drying plant oil (rapeseed oil) separately and in different mixtures. Plant oils were tested in greenhouse experiments (in vivo) on green beans (Phaseolus vulgaris L.) against bean rust (Uromyces appendiculatus). We observed that a 2% oil concentration showed no or very low phytotoxic effects on green beans. Both tested drying oils showed a protective control ranging from 53–100% for linseed oil and 32–100% for tung oil. Longer time intervals of 6 days before inoculation (6dbi) were less effective than shorter intervals of 2dbi. Curative efficacies were lower with a maximum of 51% for both oils when applied 4 days past inoculation (4dpi) with the fungus. Furthermore, the results showed no systemic effects. These results underline the potential of drying plant oils as biorationals in sustainable plant protection strategies.

RESISTANCE IN COMMON BEAN TO UROMYCES APPENDICULATUS UNDER FIELD AND GREENHOUSE CONDITIONS

Bean rust, caused by Uromyces appendiculatus, is a major disease in common bean which occurs annually in The Rhodope Mountains and sporadic in the plains of Bulgaria. The present study aims to find sources of resistance in common bean to the pathogen for using in a breeding program. The reaction of fifty-five Phaseolus vulgaris accessions to the pathogen was monitored under field condition. Infection type, disease intensity and area under the disease progress curve were calculated. Twelve common bean cultivars were inoculated with eight pathotypes of races 20-2, 20-16, and 20-18 in the greenhouse, and infection type was estimated. Twelve accessions had an immune reaction, eight accessions had resistant a reaction, two accessions had a middle resistant reaction, and seven accessions had a susceptible reaction to U. appendiculatus population in both field estimations. Five cultivars showed resistant phenotype to the eight pathotypes in the greenhouse, four of which were resistant in the field (Abritus, Beslet, Trakiya, and Prelom). Five cultivars had a susceptible or resistant reaction to the pathotypes of the same race, resulting from different interaction between resistant genes in the host and virulent genes in the pathogen. Nine accessions showed race-nonspecific resistance in the field expressed in low disease intensity and susceptible/resistant phenotype.

Benzothiadiazole Conditions the Bean Proteome for Immunity to Bean Rust

The common bean rust fungus reduces harvests of the dry, edible common bean. Natural resistance genes in the plant can provide protection until a fungal strain that breaks resistance emerges. In this study, we demonstrate that benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester (BTH) sprayed on susceptible beans induces resistance to common bean rust. Protection occurred as soon as 72 h after treatment and resulted in no signs of disease 10 days after inoculation with rust spores. By contrast, the susceptible control plants sustained heavy infections and died. To understand the effect BTH has on the bean proteome, we measured the changes of accumulation for 3,973 proteins using mass spectrometry. The set of 409 proteins with significantly increased accumulation in BTH-treated leaves included receptor-like kinases SOBIR1, CERK1, and LYK5, which perceive pathogens, and EDS1, a regulator of the salicylic acid defense pathway. Other proteins that likely contributed to resistance included pathogenesis-related proteins, a full complement of enzymes that catalyze phenylpropanoid biosynthesis, and protein receptors, transporters, and enzymes that modulate other defense responses controlled by jasmonic acid, ethylene, brassinosteroid, abscisic acid, and auxin. Increases in the accumulation of proteins required for vesicle-mediated protein secretion and RNA splicing occurred as well. By contrast, more than half of the 168 decreases belonged to chloroplast proteins and proteins involved in cell expansion. These results reveal a set of proteins needed for rust resistance and reaffirm the utility of BTH to control disease by amplifying the natural immune system of the bean plant.