White-Fringed Weevil

October 6, 2021

Sample information
Picture
Photos by: Syrah S.
Location
Collection date 08/23/2021
Captive / Cultivated? Wild-caught
Group The School for Science and Math at Vanderbilt
Observations

This white-fringed weevil was caught on the underside of a wooden mailbox. It was in the shade under a tree, in a neighborhood.
Putative identification Arthropoda Hexapoda Insecta Coleoptera Curculionidae

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

*PCR machine failed during DNA amplification for 7 test tubes; the Bordenstein lab reran our samples.

*We identified this arthropod as a white-fringed weevil based on morphology and what BLASTN results suggested, but chromatogram results for CO1 were not completely clear, therefore our identification is not conclusive.

Results
Wolbachia presence Yes
Confidence level High
Explanation of confidence level

High confidence level is due to appropriate (expected) test results from controls in both CO1 and WSPEC experiments. Positive controls were positive, negative control and water were Wolbachia negative, water was negative for arthropod DNA. There did not appear to be contamination between samples.
Wolbachia 16S sequence
GTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTGCCATCAGGTAATGCTGAGCACTTTAAGGAAACTGCCAGTGATAAGCTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCTTTATGGAGTGGGCTACACACGTGCTACAATGGTGTCTACAATGGGCTGCAAAGTCGCGAGGCTAAGCTAATCCCTAAAAAACATCTCAGTTCGGATTGTACTCTGCAACTCGAGTACATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCT
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence
TTCGTTCAGAATTAGGAAACCCNGGATCTCTGTTGGAGATGACCAAATTTATAACGTTATTGTAACTGCTCATGCTTTTA TTATAATTTTCTTTATAGTAATACCTATCATAATTGGGGGATTTGGAAACTGACTAGTTCCATTAATACTAGGAGCTCCA GACATAGCCTTCCCTCGTCTTAATAATATAAGATTTTGACTACTTCCACCTTCTTTAACTTTACTTTTAATAAGAAGAAT TGTTGATAAAGGTGCAGGAACAGGATGAACAGTNTATCCCCCTCTCTCTGCCAATGTAGCTCATGAAGGATCCTCTGTTG ATTTAGCAATTTTTANACTACATATAGCAGGTGTATCATCAATTTTAGGAGCAGTAAATTTTATTTCTACAGTTTTAAAT ATACGACCTTCAGGAATATCTCCTGATCGAATACCTTTATTTGTATGAGCAGTAGAAATTACTGCTATNTTTATTACTTT TATCTTTACCAGTTTTAGCAGGAGCAATNACTATACTTTNTAACAGATCGAAATATTAATACCTCATTTTTTGACNCCTG CAGGAGGAGGAGATCCTATTTTATANCAACATTTATTTTGATTTTTTGG
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Curculionidae was found to be postive for Wolbachia.
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