Assistance Needed: Dual Culture Assay Issues for Chilli Anthracnose disease Project
Dear Researchers,
I hope this message finds you well. I am currently working on a project focused on chilli anthracnose disease, specifically aiming to identify a potential biocontrol agent against Colletotrichum capsici using the dual culture assay by the well diffusion method.
However, I am encountering difficulties with the assay. Upon inoculating Colletotrichum capsici on PDA plates and creating wells for bacterial solutions, I am not observing any zone of inhibition. Instead, I often face issues such as contamination or no growth of the pathogen at all.
Could you kindly suggest a proper technique or approach that I could employ to improve the efficacy of the dual culture assay? Any insights or recommendations would be greatly appreciated.
Thank you in advance for your assistance.
To screen for potential biocontrol agents against Colletotrichum capsici, the causal agent of chili anthracnose, a systematic approach can be employed. The first step is to isolate and identify the pathogen by collecting infected chili samples, isolating the Colletotrichum capsici pathogen, and confirming its identity through morphological and molecular methods. Next, the in vitro screening of biocontrol agents involves obtaining potential biocontrol agents, such as beneficial microorganisms, and evaluating their antagonistic activity against the isolated pathogen using techniques like dual-culture plate assays or volatile organic compound (VOC) inhibition assays. The most effective biocontrol agents are then selected based on their ability to inhibit the growth or suppress the development of the pathogen.
Further investigations into the mechanisms of action of the selected biocontrol agents are conducted, exploring their ability to suppress the Colletotrichum capsici pathogen through antibiosis, competition for nutrients and space, induced systemic resistance, or the production of lytic enzymes. The efficacy of the biocontrol agents is then evaluated under in vivo conditions, such as greenhouse or field trials, to assess their ability to control chili anthracnose and their impact on plant health and yield. Additionally, suitable formulations of effective biocontrol agents are developed and optimized to enhance their stability, shelf-life, and ease of application. Finally, the compatibility of the biocontrol agents with other disease management strategies is evaluated, and they are integrated into an Integrated Pest Management (IPM) program for the effective and sustainable management of chili anthracnose.
Thank you for your detailed response Sakshi , now I have isolated the the pathogen and want to go for antagonistic activity with rhizosphere bacteria, can you tell me the suitable method other than dual culture assay with detail ,
my email id is [email protected]
thank you for your time Sakshi
Hi,
Screening for antifungal bacteria through simultaneous culture is likely the most effective method. Bacteria can resist fungi by producing extracellular compounds and by competing for space and nutrients. Once you have identified bacteria that can resist fungi, the next step is to use the good diffusion method.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Methods
- Cloning