Recipient Organization
AGION TECHNOLOGIES, INC.
60 AUDUBON ROAD
WAKEFIELD,MA 01880-1203
Performing Department
(N/A)
Non Technical Summary
While bacteria cause fewer plant diseases than fungi and viruses, they do cause serious economic damages to both US and worldwide agriculture. Citrus canker caused by Xanthomonas axonopodis has resulted in the destruction of over 20 million trees in Florida, while fire blight caused by Erwinia amylovora costs exceed $100 million per year in the United States alone, and recent epidemics have cost over $42 million in Michigan in 2000 and over $68 million in the Pacific Northwest in 1999. Bacterial plant pathogens are difficult to control due to shortage of chemical control agents beyond antibiotics. Traditional methods of disease control has limited effectiveness. Unfortunately, these strategies can be labor-intensive, have undesirable environmental impacts, and cost significant amounts. Perhaps because of it, apple and pear growers use antibiotics intermittently at best; studies show that growers use less than 5% of the amount of antibiotics permitted by the EPA, a mere 30,800 pounds out of an allowable potential of over 1,000,000 pounds of streptomycin. Sciessent LLC has developed bioactive systems with low levels of mixed metals that can act as very effective biocides. Sciessent believe that use of this technology in plant protection against Erwinia amylovora, Xanthomonas axonopodis pv. citri, etc. can significantly reduce the volume of antibiotics and copper in commercial disease control while also providing a valuable tool for managing the development of antibiotic-resistant organisms. Sciessent antibacterial formulations will be tested against plant pathogenic bacteria in laboratory assays, and formulations that performed well in laboratory screening, will be tested in the field. Upon completion of these goals, Sciessent will have several antimicrobial formulations that are sufficiently developed for Phase II testing, that include waterfastness, effectiveness against bacterial pathogens outside the scope of Phase I testing, stability, and lowest feasible application rates.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Goals / Objectives
The goal of Phase I is to discern which of existing Sciessent formulations provides the highest levels of effectiveness against the plant pathogenic bacterial pathogens. Upon completion of this goal, Sciessent will have several antimicrobial formulations that are sufficiently developed for Phase II testing that includes waterfastness, effectiveness against bacterial pathogens outside the scope of Phase I testing, stability, and lowest feasible application rates. To achieve the Phase I goal, Sciessent has two technical objectives for this eight month grant term (May 15, 2011 - January 15, 2011). 1. Screen Sciessent antibacterial formulations against plant pathogenic bacteria Xanthomonas campestris, Calvibacter michiganensis, Pseudomonas syringae pv. tomato and Erwinia amylovora in laboratory assays, and 2. Test the Sciessent formulations that performed well (providing 100 percent inhibition of growth in laboratory screening) in the field at different locations. The long-range output of Sciessent's efforts to develop microbiocides for agricultural use will be the development of an effective microbiocide for use against Erwinia amylovora, Xanthomonas campestris, Pseudomonas syringae pv. tomato and Clavibacter michganensis pv. michiganensis. The immediate outputs of Phase I testing will be: 1. Data documenting the in laboratory effectiveness of Sciessent's six silver-ion microbiocide formulations against the following plant pathogenic bacteria: Xanthomonas campestris, Calvibacter michiganensis, Pseudomonas syringae pv. tomato and Erwinia amylovora; 2. Formulation(s) of Sciessent microbiocides suitable for field testing; 3. Data documenting the field testing effectiveness of the formulation(s) against any or all of the following plant pathogenic bacteria: Xanthomonas campestris, Calvibacter michiganensis, Pseudomonas syringae pv. tomato and Erwinia amylovora; and 4. Formulation(s) of Sciessent microbiocides recommended for further Phase II testing and development.
Project Methods
Objective 1: The Minimum Inhibitory Concentration (MIC) will be determined for the bacteria including Xanthomonas campestris pv. vesicatoria, Calvibacter michiganensis, Pseudomonas syringae pv. tomato and Erwinia amylovora as one of the following processes. 1. Amending the nutrient agar medium or synthetic minimal agar medium with concentrations ranging from 10 ppm to 200 ppm of the Agion formulations before pouring it in the Petri plates and then spreading 0.1 ml of the test bacterial suspension (10,000/ml) and incubating it at 25 C in dark for 24 hours and recording the growth of bacterial colonies, or 2. Amending the 5ml liquid nutrient broth medium or synthetic minimal broth medium with concentrations ranging from 10 ppm to 200 ppm of the formulations and then adding 0.0l ml of 100,000/ml bacterial cells in them and incubating at 20 C and monitoring their growth over time for 48 hours. Growth over time will be measured by measuring turbidity through optical density and also dilution plating them on Tryptic Soy Agar (TSA) or nutrient agar medium to obtain comparable cfu counts. There will be five replications in a completely randomized block design. Statistical analysis, ANOVA and mean separations, etc. will be conducted using software JMP 8 or equivalent statistical program. Objective 2: Since Sciessent does not have field testing facilities, collaborators at University of Illinois,Michigan State University and LABS Services will conduct field tests under the protocols developed by Project Director Dr. Naseem Khan For Fire Blight of apples, there will be three to four replications with at least one tree per replication in a completely randomized block design. Formulations will be applied at 80 percent to full bloom at 300 gallon per acre rate with a hand gun at 350-400 psi. Trees will then be inoculated with Erwinia amylovora 24 hours after the formulation application, in the evening, at a concentration of 5.0X106 cfu per ml. Fire blight spur infection will be evaluated 5 days after by determining the percentage of spurs infected and percent of blossom blight caused by E. amylovora from at least 200 terminals samples. Treatments will include a non-treated and a standard antibiotic control. Data collected on percent disease severity and percent disease incidence will then be transformed to arcsine before statistical analysis including ANOVA and mean separation, etc., is performed. Statistical analysis, ANOVA and mean separations, etc. will be conducted using software JMP 8 or similar statistical program. Means will be separated by Tukey-Kramer HSD procedure. For Tomato Bacterial Leaf Spot, Bacterial Speck and Bacterial Canker, there will be three to four replications in a completely randomized block design. Formulations will be applied at first sign of disease observed and will continue with 5-7 days interval. Plants will be observed daily for disease signs and symptoms. Percent disease and disease incidence will be recorded when disease in untreated controls exceed 50%. Statistical analysis, ANOVA and mean separations, etc. will be conducted using software JMP 8 or equivalent statistical program.