Surrogate Host Bacteriophage Propagation for Plant Disease Control
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Solution Overview
Problem
Current methods for propagating bacteriophages capable of infecting Xylella fastidiosa and Xanthomonas axonopodis are labor-intensive and inefficient, requiring complex media and long durations, making large-scale production and treatment of plant diseases challenging.
Innovation Solution
A method involving the use of a surrogate host, such as Xanthomonas species EC-12, to rapidly propagate and isolate virulent bacteriophages that can infect and lyse Xylella fastidiosa and Xanthomonas axonopodis, using a soft agar overlay and CsCl purification to achieve high-titer phage lysates, which can be stored and used for treating plant diseases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used to propagate bacteriophages capable of infecting Xylella fastidiosa, then the bacteriophages can be produced, but the process is labor-intensive and inefficient requiring complex media and long durations
Solution Approach 1:
The patent uses a surrogate host (Xanthomonas axonopodis) as an intermediary organism to propagate bacteriophages that ultimately infect Xylella fastidiosa. The surrogate host serves as a mediator that enables efficient phage production without requiring complex growth conditions, thereby resolving the contradiction between productivity and media complexity
Solution Approach 2:
The invention changes the host organism parameter from the target pathogen (Xylella fastidiosa) to a surrogate host (Xanthomonas axonopodis) that has more favorable growth characteristics. This parameter change enables rapid bacteriophage propagation in simple media, dramatically improving productivity while reducing complexity
2Productivity
If conventional methods are used to propagate bacteriophages, then bacteriophages can be produced, but the process requires long durations
Solution Approach 1:
The surrogate host acts as a temporal mediator that accelerates the bacteriophage propagation process. By using an organism with faster generation time and more rapid phage replication kinetics, the overall propagation duration is dramatically reduced while maintaining high productivity
Solution Approach 2:
The method performs preliminary propagation of bacteriophages in the surrogate host system before actual treatment application. This preliminary action in an optimized system prepares high-titer phage stocks rapidly, reducing the time required for subsequent treatment preparations
3Quantity of substance
If large-scale production of bacteriophages is attempted using conventional methods, then treatment of plant diseases can be pursued, but the process becomes impractical due to labor intensity and inefficiency
Solution Approach 1:
The surrogate host system serves as a scalable intermediary platform that enables large-scale bacteriophage production through simple, repeatable procedures. The system allows straightforward manipulation and scaling without the operational complexities associated with culturing the actual pathogen at large scales
Solution Approach 2:
The method employs disposable surrogate host cultures that can be rapidly prepared and discarded after phage propagation. This approach eliminates the need for maintaining complex long-term cultures, simplifying operations and enabling easy scaling by simply preparing additional disposable culture batches
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for cost-effective and efficient production of bacteriophages that can effectively reduce or prevent symptoms of Pierce's Disease and citrus canker in plants, providing a viable biocontrol option by rapidly producing high-titer phage lysates that can be used for treatment.
Implementation Method 1
isolating at least a first bacteriophage from the population capable of lysing said Xylella fastidiosa and Xanthomonas axonopodis
Implementation Method 2
using a soft agar overlay
Implementation Method 3
CsCl purification to achieve high-titer phage lysates
Data Source
AI summary
Methods and compositions are provided for preventing or reducing symptoms or disease associated with Xylella fastidiosa or Xanthomonas axonopodis in a plant. The invention provides novel bacteriophages virulent to Xylella fastidiosa or Xanthomonas axonopodis, including XfaMija and XfaMijo, and further provides methods for treating or preventing Pierce's Disease or Citrus Canker in plants.