Progress 06/01/17 to 01/31/19
Outputs
Target Audience:During the project period APSE carried out research to demonstrate the effectiveness of RNAi in killing red fire ant workers and 4th instar larvae. The resulting data from this study is of interest toresearchers in academia and industry working on developing applications of RNAi for insect control. The experimental results are also expected to provide impetus to business leaders working towards developing sustainable, nontoxic and benign pest control products. The data from this research provides proof of concept for application of RNAi for fire ant control and will help APSE in securing additional funding/collaboration with major players in pesticides and pest control business. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Results of the fire ant bioassay were reported in various public conferences and in many private presentations to strategic industry players as well as various venture capital concerns. Each of these interactions provided opportunities for company people to layout the details of APSE's plan for fire ant control and be challenged by people knowledgeable in the business/technology. How have the results been disseminated to communities of interest?Theresearch data from this research effort was disseminated through various public conferences and by private presentations to strategic industry players as well as various venture capital concerns with the goal of attracting additional funding or establishing strategic collaborations. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
During the project periodall the three objectives of the project as described above were completed. APSE employed bioinformatics tools to compare RIFA genome and transcriptome against sequences from other ant species to identify 12 RNAi target sequences specific to RIFA that target 7 different genes. The selected sequences were cloned into APSE's RNA production vector. The rnc gene knockout strain of E.coli (HT115(DE3)) was transformed with the RNA production vector carrying RIFA sequences to produce dsRNA for bioassay on RIFA workers. Two bioassays on RIFA workers wereconducted to test the efficacy of dsRNA targeting selected target genes. Based on the results of the bioassays, four dsRNA treatments targeting different RIFA genes were identified for further testing. Two of the selected dsRNA treatments were tested in a feeding bioassay on RIFA 4th instar larvae. The 4th instar larvae play a very important role in RIFA colonies because it is the only life stage that can digest solid food and provides nutrients for queen/s and other workers in a RIFA colony. The ability of any product to reach RIFA 4th instar larvae (food/products are delivered by workers to larvae) without degradation and then be effective in killing the 4th instar larvae is key towards developing a colony scale control. Both dsRNA treatments were found to be lethal to 4th instar larvae as less than 3% of the treated larvae progressed to pupal stage. RNA targets were also found to be more effective than commercial control (Advion ant gel) in killing RIFA workers and 4th instar larvae. Moreover, the mortality rate observed with the non-specific dsRNA treatment was similar to the no RNA control (10% sucrose treatment) in both RIFA worker and 4th instar larvae bioassays, pointing towards insignificant effects of non-specific RNA on the test individuals. These finding are important as they demonstrate the lethality of the selected RNAi gene targets to RIFA and can pave the way for developing an RNAi based environmentally friendly, target specific product to limit RIFA infestation. With the successful completion of all phase1 objectives, APSE is very confident that an RNAi based product limiting RIFA spread can be developed. Successful development of an RNAi based control for RIFA would have wide ranging impacts on the environment and biodiversity of the infested area and in limiting annual loses to crops, farmed animals, infrastructure etc. estimated to be around $6.3 B in continental US and Puerto Rico.
Publications
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Progress 06/01/17 to 01/31/18
Outputs
Target Audience:During the project period APSE carried out research to demonstrate the effectiveness of RNAi in killing fire ant workers. The resulting data from this study is of interest toresearchers in academia and industry working on developing applications of RNAi for insect control. The experimental results are also expected to provide impetus to business leaders working towards developing sustainable, nontoxic and benign pest control products. The data from this research provides proof of concept for application of RNAi for fire ant control and will help APSE in securing additional funding/collaboration with major players in pesticides and pest control business. Changes/Problems:On set of winter during the project period prevented APSE from completing the experiments proposed under objective 3. APSE has been granted no cost extension to complete all the experiments proposed under objective 3. What opportunities for training and professional development has the project provided?Results of the fire ant bioassay were reported in various public conferences and in many private presentations to strategic industry players as well as various venture capital concerns. Each of these interactions provided opportunities for company people to layout the details of APSE's plan for fire ant control and be challenged by people knowledgeable in the business/technology. How have the results been disseminated to communities of interest?The preliminary research data from this reaearch effort was disseminated through various public conferences and by private presentations to strategic industry players as well as various venture capital concerns with the goal of attracting additional funding or establishing strategic collaborations. What do you plan to do during the next reporting period to accomplish the goals?This is the final report for this project. However, APSE has been granted no cost extension to complete the experiments proposed under objective 3 of this project. APSE was unable to complete objective 3 of theproject during the scheduled project period due to the onset of winter. Feeding biossays on RIFA are not carried out during winter due to changes in the feeding behavior and energy requirements of a RIFA colony. APSE will complete all the experiments proposed under objective 3 during the no cost extension period and report the progress as required by NIFA.
Impacts
What was accomplished under these goals?
During the project period the first two objectives of the project as described above were completed. APSE employed bioinformatics tools to compare RIFA genome and transcriptome against sequences from other ant species to identify 12 RNAi target sequences specific to RIFA that target 7 different genes. The selected sequences were cloned into APSE's RNA production vector. The rnc gene knockout strain of E.coli (HT115(DE3)) was transformed with the RNA production vector carrying RIFA sequences to produce dsRNA for bioassay on RIFA workers. A bioassay on RIFA workers was conducted to test the efficacy of selected target sequences. Based on the results of the bioassay, two sequences targeting two different RIFA genes have been identified for further testing. These selected sequences will be tested on RIFA 4th instar larvae. APSE was unable to complete the third objective during the project period due to the onset of winter. Feeding biossays on RIFA are not carried out during winter due to changes in the feeding behavior and energy requirements of a RIFA colony. APSE has been granted no cost extensionto complete the third objective.
Publications
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Progress 06/01/17 to 01/31/18
Outputs
Target Audience:During this reporting period APSE carried out research to demonstrate the effectiveness of RNAi in controlling/killing fire ant workers. The data generated from the research during this period is of interest for researchers in academia and industry working on developing applications of RNAi. The experimental results will also provideimpetusto business leaders working towards developing sustainable, nontoxic and benign pest control products.The data from research serves as a proof of concept for application of RNAi for fire ant control and will help APSE in securing additional funding/ collaboration with major players in pesticides and pest control business. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Interim results of the fire ant testing were reported in various public conferences and in many private presentations to strategic industry players as well as various venture capital concerns. Each of these provide opportunites of company people to lay out the details of our plans for fire ant control and be challenged by people knowledgeable in the business/technology. How have the results been disseminated to communities of interest?The preliminary research data was disseminatedthroughvarious public conferences and by private presentations to strategic industry players as well as various venture capital concerns with the goal ofattracting additional funding or establishing strategic collaborations. What do you plan to do during the next reporting period to accomplish the goals?Experiments will be carried out during the next reporting period to test the efficacy of the two selected RIFA gene targets in killing the 4th instar larvae.
Impacts
What was accomplished under these goals?
The first two objectives of the project as described above were completed during this reporting period. APSE used bioinformatics tools to compare RIFA genome and transcriptome against sequences from other ant species to identify 12 RNAi target sequences specific to RIFA that target 7 different RIFA genes. The selected sequences were cloned into APSE's RNA production vector. The rnc gene knockout strain of E.coli (HT115(DE3)) was transformed with the RNA production vector carrying RIFA sequences to produce dsRNA for bioassay on RIFA workers. A bioassay on RIFA workers was conducted to test the efficacy of selected target sequences. Based on the bioassay two sequences targeting two different RIFA genes have been identified for further testing. These selected sequences will be tested on RIFA 4th instar larvae.
Publications
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