Title

Barcoding Adelpha immatures to understand host breadth

Lead Author Major

Connor Soderstrom

Lead Author Status

5th year Senior

Second Author Major

Elizabeth Song

Second Author Status

Junior

Third Author Major

Rylie Towne

Third Author Status

5th year Senior

Format

Poster Presentation

Faculty Mentor Name

Dr. Ryan Hill

Faculty Mentor Department

Dept of Biological Sciences

Abstract/Artist Statement

With more than 200 taxa, Adelpha butterflies are useful for examining evolution of biological diversification. For example, the immense diversity in Adelpha is hypothesized to result from relationships with a wide diversity in larval hostplants. After extensive immature-stage fieldwork in Costa Rica and Ecuador, we turned to DNA barcoding using cytochrome oxidase subunit I (CoI) to identify immatures for several reasons: 1) mortality during rearing was as high as 33%, leaving some Adelpha-host interactions undetermined; 2) deceased larvae tend to be on hosts that were used by multiple species, and identifying them could increase sample size or represent new Adelpha-host interactions; 3) in Ecuador, field work uncovered Adelpha species using many new host genera, including rarer Adepha with low sample size. Given the high diversity of Adelpha species and hosts in Ecuador it is likely some of the deceased larvae represent new Adelpha-host interactions. Amplification was successful for egg, larval and pupal tissue. However, eggs and pupae had lower success because of parasitism/degradation before preservation. Several other issues arose making some, but not all, identifications challenging. First, a relatively high 70.9%(39/55) of larval samples failed to amplify at first. To test whether host plant chemistry such as polyphenols could be inhibiting PCR, we cleaned the genomic DNA and re-amplified with 69.2%(27/39) success. Second, the BOLD and GenBank databases produced conflicting identifications (i.e.>0.5% diff.) in 40.2% of samples. Third, the databases had relatively few sequences from South America, limiting our identification to <98% match in BOLD and GenBank in 16.5% and 51.2% of samples, respectively. Finally, some species in Ecuador cannot be identified with CoI, similar to Costa Rica. Despite these challenges, CoI appears useful for identifying many Adelpha-host interactions, helping clarify patterns in this diverse lineage.

Location

Virtual

Start Date

25-4-2020 1:00 PM

End Date

25-4-2020 3:00 PM

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Apr 25th, 1:00 PM Apr 25th, 3:00 PM

Barcoding Adelpha immatures to understand host breadth

Virtual

With more than 200 taxa, Adelpha butterflies are useful for examining evolution of biological diversification. For example, the immense diversity in Adelpha is hypothesized to result from relationships with a wide diversity in larval hostplants. After extensive immature-stage fieldwork in Costa Rica and Ecuador, we turned to DNA barcoding using cytochrome oxidase subunit I (CoI) to identify immatures for several reasons: 1) mortality during rearing was as high as 33%, leaving some Adelpha-host interactions undetermined; 2) deceased larvae tend to be on hosts that were used by multiple species, and identifying them could increase sample size or represent new Adelpha-host interactions; 3) in Ecuador, field work uncovered Adelpha species using many new host genera, including rarer Adepha with low sample size. Given the high diversity of Adelpha species and hosts in Ecuador it is likely some of the deceased larvae represent new Adelpha-host interactions. Amplification was successful for egg, larval and pupal tissue. However, eggs and pupae had lower success because of parasitism/degradation before preservation. Several other issues arose making some, but not all, identifications challenging. First, a relatively high 70.9%(39/55) of larval samples failed to amplify at first. To test whether host plant chemistry such as polyphenols could be inhibiting PCR, we cleaned the genomic DNA and re-amplified with 69.2%(27/39) success. Second, the BOLD and GenBank databases produced conflicting identifications (i.e.>0.5% diff.) in 40.2% of samples. Third, the databases had relatively few sequences from South America, limiting our identification to <98% match in BOLD and GenBank in 16.5% and 51.2% of samples, respectively. Finally, some species in Ecuador cannot be identified with CoI, similar to Costa Rica. Despite these challenges, CoI appears useful for identifying many Adelpha-host interactions, helping clarify patterns in this diverse lineage.