Biocontrol of bean bacterial wilt disease using Pseudomonas fluorescens and Trichoderma harzianum biological agents

Document Type : Research paper-Persian

Authors

1 Assistant Professor, Department of Plant Protection, Faculty of Agriculture, University of Lorestan, Khorramabad, Iran

2 Ph.D. student, Department of Plant Protection, Faculty of Agriculture, University of Lorestan, Khorramabad, Iran

3 Instructor, Department of Agricultural Science, Technical and Vocational University (TVU), Tehran, Iran

4 Ph.D. student, Department of Plant Protection, Faculty of agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Background and Objectives
Curtobacterium flaccumfaciens pv. flaccumfaciens is the cause of bean bacterial wilt (CFF) and a dangerous pathogen for bean fields. To date, no definitive control has been found for this pathogen. In sustainable agriculture, it is used to control plant pathogens by applying management methods related to preserving the environment and producing organic products. The use of biological agents such as Trichoderma fungi and Pseudomonas bacteria is among these management methods. Minimal research has been done worldwide on biological agents' effects on CFF. Therefore, the present research will investigate the effect of inhibiting the growth and activity of two biological agents‌, including Pseudomonas fluorescens and Trichoderma harzianum, against CFF in laboratory and greenhouse conditions.

Materials and Methods

First, the pathogenicity test of CFF was carried out on the greenhouse's Sadri variety pinto bean plant. Then, the effect of fungal and bacterial biological agents against the pathogenic agent was investigated by two methods of mutual culture and their secreted substances in laboratory conditions. By measuring the diameter of the transparent halo formed around the extract and spot culture of biological agents, the growth inhibition power of these biological agents against CFF was evaluated. In order to investigate the effect of these biological agents against bacterial wilt disease in greenhouse conditions, these biological agents and pathogens were propagated and expanded, and a standard suspension was first prepared from them. These suspensions were inoculated on the host plant using the spraying method. During 21 days, disease symptoms and bean plant indicators have been recorded. Then, the disease severity of progress under the influence of these biological factors was checked using the 0 to 4 scoring system. Also, the plant resistance type to the pathogen was evaluated using the 1-5 scoring system. The data obtained from the laboratory (inhibition percentage) and greenhouse (treatment changes, plant resistance, and plant indices) studies were analyzed in a completely randomized design using SPSS 21 software.

Results

In laboratory conditions, P. fluorescens and T. harzianum treatment prevented the growth of pathogenic bacteria CFF by 18.67 and 27.33%, with a diameter of the inhibitory halo of 14 and 20.5 mm, respectively. It was also found that a significant part of the inhibitory effect of P. fluorescens bacteria (about 85.70%) is related to the secreted substances of the bacterial cells. In contrast, in the biological fungus T. harzianum, the effect of the secreted substances was about 50.02%. The effect of biological factors on CFF in greenhouse conditions showed that these biological factors strengthen the defense reaction of bean plants against CFF. So, the sensitive reaction of the plant against CFF bacteria (with a disease severity of 83.13%) was transformed into a semi-resistant reaction by inoculating P. fluorescens and T. harzianum with the diseased plant. Also, the inoculation of the combination of these two biological factors made the plant resistant to pathogenic bacteria. Treatment of P. fluorescens+T. harzianum, by affecting CFF bacteria, was the best treatment in reducing the severity of the disease during 7, 14, and 21 days; the severity of the disease on these days was 21.87, 33.12, and 36.25%, respectively, which showed a significant difference with the disease severity index in infected treatment during these few days at the statistical level of 1%. Treatment of P. fluorescens+T. harzianum, with a disease severity index of 36.25% and a 57.04% reduction in disease severity, showed the most significant effect on the reduction of bean bacterial wilt disease.

Discussion

In the present study, the use of two biological agents, P. fluorescens and T. harzianum, showed significant controlling effects on CFF in laboratory and greenhouse conditions, and 57.04% reduced the severity of this disease in the bean plant. These two biological factors prevent the growth and spread of plant pathogens by using mechanisms such as the production of secondary metabolites, acidifying the environment, competing for food and space, colonizing and stimulating the plant to produce phytoalexin, proteins related to pathogens, salicylic acid, etc. Therefore, using these two biological agents as safe, effective, and durable biological control agents in sustainable agriculture against bean bacterial wilt disease is recommended.

Keywords

Main Subjects


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