Progress 05/01/23 to 12/04/24
Outputs
Target Audience:These activities reached academic and industry scientific communities and members of the public. Changes/Problems:Onesignificant changeis the transition of the PD from a postdoctoral positionto a tenure track faculty position. The last date of work on this project by the PD as a postdoctoral researcher at UMD was 8/29/24. As that date is only one month after the submission of the last annual report, much of the progress reported in this 'final report'was also reported in the annual report. The PD intends to continue work on this research project in her new research lab. What opportunities for training and professional development has the project provided?An undergraduate student from a historically underrepresented group was recruited for a summer research experience in the agricultural sciences. The student was provided career and research mentorship. The student completed an independent research project and developed skills in bioinformatics and genetic analysis. The student has produced a scientific poster related to the goals in this proposal, which will be presented at a scientific meeting next year. The career development objectives of this grant for the PD have been met. Over the last year the PD developed materials for the academic job market, and applied, interviewed, and accepted a tenure track faculty position with teaching and research components. She has now begun establishing her research lab at Hofstra University. The PD also developed new skills in chemical ecology including insect headspace capture and analysis and the identification and curation of gene sequences important to insect olfaction. She has also developed as a mentor and teacherthrough the recruitment and training of an undergraduate mentee. She developed skills in teaching big data science by completing a week-long instructor training program through the data/software carpentry program and learned about current best practices in undergraduate teaching by attending the Innovations in Teaching and Learning Conference at UMD. How have the results been disseminated to communities of interest?The PD attended andpresented at the national meeting of the Entomological Society of America. She shared her work in both an oral and poster presentation and discussedthis project with multiple scientists from across the US. She also will share her work at two additionalconferences in the next year. The PD also engaged with the public on the topic of entomology by participating in the Maryland Day outreach activities about insects. She also assisted with the production of an educational video from PBS digital studios about lacewing biology. What do you plan to do during the next reporting period to accomplish the goals?In the analysis of data gathered for objective 1, we identified some potential contaminant compounds. We have a new protocol to better remove potential contaminants which we plan to use during a second round of data collection during the next reporting period. That new analysis should produce quality data and more replicates to expand and strengthen the work related to objective 1. To expand our characterization of lacewing odorant binding protein sequences to moreChrysoperlaspecies (objective 2) we will sequence and analyze genetic data from samples collected this year. To follow up on our results from objective 3, we plan to measure binding affinity of the volatiles we collected in lacewing headspace to the identified lacewing OBP sequences.
Impacts
What was accomplished under these goals?
We have characterized thevolatiles produced bylacewings in the genus Chrysoperla. We collected andidentified headspace volatile compounds from multiple species and both sexes (Objective 1.1). By analyzing multiple species and sexes we have started to characterize the extent to which volatile profiles vary by sex and species (Objective 1.2). From this analysis we were able toidentifypromising candidate compounds, which were not previously known to be involved in lacewing chemosensation. These candidate compoundsare present in the headspace of lacewings and similar to known pheromones in other insect species. Wealso identified other compounds in the headspace with unknown function, which might be revealed byfurther study. We then expanded upon the findingsfrom the analyses in Objective 1.We validatedthe relevance of the identified compounds using behavioral analysis. With a y-tube assay we compared thevolatile profile produced by a male or female lacewing to a blank negative control, and showed that conspecifics move more often towards the odor of a lacewing. This result suggests that the volatilesidentified for objective 1 are biologically relevant. We have started to characterize the diversity of OBP inChrysoperlausing publicly available data. In this analysis we were able to characterize the evolution and diversity of the odorant binding proteins offour Chrysoperla species(Objective 2.2). We have manually curated this dataset to confirm gene model quality and identifiedlikely OBPs missed by automated annotation. For thesespecies we have characterized the number, genomic positions, and evolutionary trajectory of OBPs in lacewings (Objective 2.3). We also collected samples from additional species which have been preserved for sequencing oncethe grant is transferred to the new institution of the PD(Objective 2.1). We have also tested how simulated bindingaffinitiescompareto known bindingaffinitiesfrom the published literature (3.1).In our analysis the volatiles that truly bind to OBP1 had higher binding affinity using our computational approach, which is encouraging proof of concept for the potential of molecular docking. However, in our analysis the volatiles that previous studies showed did not bind to OBP1 had a range of binding affinities, some high some low. These results suggest that this approach is not a silver bullet and is likely to yield both true and false positives.
Publications
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Progress 05/01/23 to 04/30/24
Outputs
Target Audience:Activities reached academic and industry scientific communities and members of the public. Changes/Problems:Originally, we planned to send our samples out for headspace analysis (objective 1), but an opportunity arose through collaboration to use a state-of-the-art GCMS for only the cost of supplies. This alternative analysis route gave the PD the opportunity to learn about and be involved in all stages of data collection. This also significantly decreased the cost per sample. What opportunities for training and professional development has the project provided?An undergraduate student from a historically underrepresented group was recruited for a summer research experience in the agricultural sciences. The student was provided career and research mentorship. The student completed an independent research project and developed skills in bioinformatics and genetic analysis. The student has produced a scientific poster related to the goals of this project, which will be presented at a scientific meeting in the next year. The career development objectives of this grant for the PD have been met. Over the last year the PD developed materials for the academic job market, and applied, interviewed, and accepted a tenure track faculty position with teaching and research components. The PD also developed new skills in chemical ecology including insect headspace capture and analysis and the identification and curation of gene sequences important to insect olfaction. She has also developed as a mentor and teacher through the recruitment and training of an undergraduate mentee. She developed skills in teaching big data science by completing a week-long instructor training program through the data/software carpentry program and learned about current best practices in undergraduate teaching by attending an innovations in teaching and learning conference. How have the results been disseminated to communities of interest?The PD attended and presented at the national meeting of the Entomological Society of America. She shared her work in both an oral and poster presentation and discussedthis project with multiple scientists from across the US. The PD also engaged with the public on the topic of entomology by participating in the Maryland Day outreach activities about insects. What do you plan to do during the next reporting period to accomplish the goals?In the analysis of data gathered for objective 1, we identified some potential contaminant compounds. We have a new protocol to better remove potential contaminants which we plan to use during a second round of data collection during the next reporting period. That new analysis should produce quality data and more replicates to expand and strengthen the work related to objective 1. To expand our characterization of lacewing odorant binding protein sequences to more Chrysoperla species (objective 2) we will sequence and analyze genetic data from the samples collected this year. To follow up on our results from objective 3, we plan to measure binding affinity of the volatiles we collected in lacewing headspace to the identified lacewing OBP sequences.
Impacts
What was accomplished under these goals?
We identified volatiles produced by Chrysoperla lacewings by collecting and identifying headspace volatile compounds from multiple species and both sexes (Objective 1.1). By analyzing multiple species and sexes we have started to characterize the extent to which volatile profiles vary by sex and species (Objective 1.2). We have already identified promising candidate compounds, which have not previously been reported to be involved in lacewing chemosensation. These candidates are present in the headspace of lacewings and similar to known pheromones in other insect species. We additionally identified other compounds in the headspace with unknow function which further study may reveal.To improve impact of the publication resulting from the analyses for Objective 1, we also sought to validate the relevance of the identified compounds using behavioral analysis. Using a y-tube assay we compared the full volatile profile produced by a male or female lacewing to a blank negative control, and showed that conspecifics move more often towards the odor of a lacewing. This result suggests that the volatile profiles identified for objective 1 are biologically relevant. We have started to characterize the diversity of OBP in Chrysoperla using publicly available data. In this preliminary analysis we were able to characterize the evolution and diversity of the odorant binding proteins offour Chrysoperla species (Objective 2.2). We have manually curated this dataset to confirm gene model quality and identifiedlikely OBPs missed by automated annotation. For these species we have characterized the number, genomic positions, and evolutionary trajectory of OBPs in lacewings (Objective 2.3). We collected samples from additional species which have been preserved for OBPanalysis in the next reporting period(Objective 2.1). We have also tested how simulated bindingaffinitiescompareto known bindingaffinitiesfrom the published literature (3.1). In our analysis the volatiles that truly bind to OBP1 had higher binding affinity using this computational approach, which is encouraging proof of concept for the potential of molecular docking. However, in our analysis the volatiles that previous studies showed did not bind to OBP1 had a range of binding affinities, some high some low. These results suggest that this approach is not a silver bullet and is likely to yield both true and false positives.
Publications
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