Lycorma delicatula JA02

June 9, 2022

Sample information
Picture
Photos by: Jules A.
Location
Collection date 09/27/2021
Captive / Cultivated? Wild-caught
Group Pingry School
Observations

spotted wings, orange-red wings, found by trees in a grassy area in fall 2021 and frozen at -80 degrees Celsius
Putative identification Arthropoda Hexapoda Insecta Hemiptera Fulgoridae Lycorma Lycorma delicatula

Methods
Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system Standard electrophoresis system
Buffer TAE
DNA stain SYBR Safe
Gel images
Protocol notes

Lane 1 – Ladder

Lane 2 – Lycorma delicatula Arthropod 1x dilution (COI)

Lane 3 – Lycorma delicatula Arthropod 10x dilution (COI)

Lane 4 – Lycorma delicatula Wolbachia 1x dilution (16s)

Lane 5 – Lycorma delicatula Wolbachia 10x dilution (16s)

Band in lane two at 708 bp shows that the Lycorma delicatula is an arthropod and the band in lane four at 438 bp shows the Lycorma delicatula has Wolbachia. Other lanes are other dilutions and a different bug and the protocol ran without controls.

Results
Wolbachia presence Yes
Confidence level High
Explanation of confidence level

The gel showed a band at 438 bp corresponding to the Wolbachia strain. Additionally, the DNA sequence from the Lycorma delicatula matched an uncultured bacterium found in 2011(Vanderbilt, GenBank: JN595428.1, Nasonia). The DNA also matched some Wolbachia endosymbionts at 100% in further entries.
Wolbachia 16S sequence Download FASTA    Download AB1
ATTCGATTAGAATTAATACAACCAGGATCAATAATTAAAAATGACCAAATCTATAATACCATCGTAACATCTCATGCATTCATTATAATTTTCTTTATAGTTATACCTATTATAATTGGAGGATTTGGAAACTGATTAGTTCCTATAATAATCGGTGCACCAGACATGGCATTCCCACGAATAAATAATATAAGATTTTGATTACTACCACCATCAATTTCACTTTTAATTGCAGGATCAATCGTAGGGTCAGGATCAGGTACAGGATGAACTGTTTACCCCCCTCTCTCAGCCCAAATTGCACATTCAGGACCTTCAGTAGACCTAACTATTTTTTCACTTCACATCGCAGGTTTAAGATCAATCATAGGAGCAATCAATTTCATTTCAACAACAATAAATATACGGCCAAAAGGAATAACTATAGAAAAAATACCCCTTTTCTGTTGATCAGTCCTAATTACAGCAGTTTTACTACTAGTATCATTACCAGTACTAGCTGGAGCAATTACAATACTACTAACAGATCGAAACTTTAATACCTCATTTTTTGACCCCACGGGTGGAGGAGACCCAATTTTATATCAACACTTATTCTGATTTTTTG
BLAST at The Wolbachia Project   BLAST at NCBI
Arthropod COI sequence Download FASTA    Download AB1
CGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCATCCTTAGTTACCATCAGGTAATGCTGGGGACTTTAAGGAGACTGCCAGTGATAAACTGGAGGAAGGTGGGGATGATGTCAAGTCATCATGGCCCTTATGGAGTGGGCTACACACGTGCTACAATGGTGGCTACAATGGGCTGCAAAGTCGCGAGGCTAAGCCAATCCCTTAAAAGCCATCTCAGTTCGGATTGTACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCACGCCACGGTGAATACGTTCTCGGGTCTTGTACACACTGCCCGTCACGCCATGGGAATTGGT
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Lycorma delicatula was found to be postive for Wolbachia.
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