Drosophila ananassae (Fruit Fly)

Sample information

Picture
Photos by: Kalena K.
Location
Collection date 09/06/2023
Captive / Cultivated? Wild-caught
Group Hawaii Community College
Observations

All flies from both groups displayed attraction to the rotten fruit samples, particularly the Waiwi, and were more responsive as well as active in the morning and early evening/afternoon. They were the least active and less reactive to light at night.

Putative identification Arthropoda Hexapoda Insecta Diptera Drosophilidae Drosophila

Methods

Extraction kit Edwards Buffer
DNA extraction location Whole arthropod
Single or Duplex PCR Single Reaction
Gel electrophoresis system Edvotek Gel Electrophoresis
Buffer TBE
DNA stain SYBR Safe
Gel images
Protocol notes

Three arthropods were selected as a low-elevation sample group, all three were included in the protocol.

Results

Wolbachia presence Yes
Confidence level High
Explanation of confidence level

All PCR control were successful, Drosophilinae samples (2a-7a), (+) DNA control (8a), (+) Drosophila control (2b), and (-) Drosophila control (3b) had bands at 708 bp, positive results of arthropod barcoding gene to confirm presence of CO1 gene. The three Drosophilinae samples from Low Elevation (5c-7c), (+) DNA control (8c), and (+) Drosophila control (6b) all had bands at 438 bp, positive results of Wolbachia specific gene to find the presence of 16S rRNA (Figure 1). The data from each of the controls tells me that this experiment went as expected, DNA was present where anticipated and Wolbachia was present where assumed so the arthropods tested are confidently infected with Wolbachia. Therefore, the gel electrophoresis results disprove my original hypothesis that there would be no difference between the Wolbachia infection prevalence of fruit flies in a higher elevation versus a lower elevation. There is a difference between the Wolbachia infection prevalence of Drosophilinae in a higher elevation versus a lower elevation, the lower elevation Drosophilinae samples were all positive unlike the high elevation which were all negative. 

Wolbachia 16S sequence Download AB1
Arthropod COI sequence Download AB1
>Drosophila ananassae TTTTGGCTTGAGCTGGAATAGTTGGAACTTCACTAAGTATTTTAATTCGAGCTGAATTAGGACACCCTGGA GCTTTAATTGGAGATGATCAAATTTATAACGTTATTGTAACAGCACACGCTTTTATTATAATTTTTTTCATGGTTATACC AATTATAATTGGAGGATTTGGGAATTGATTAGTTCCTTTAATATTAGGAGCACCTGATATAGCATTCCCACGAATAAATA ATATAAGATTTTGATTACTACCCCCTGCTCTTTCTCTATTATTAGTAAGAAGAATAGTTGAAAATGGAGCTGGTACTGGG TGAACAGTTTACCCACCTCTTTCAGCTGGAATTGCTCATGGAGGGGCTTCAGTTGATCTAGCTATTTTTTCATTACATTT AGCCGGAATTTCTTCAATTTTAGGAGCTGTAAATTTTATTACAACAGTAATTAATATACGATCAACTGGAATTACTCTAG ATCGTATACCTTTATTTGTTTGATCGGTAGTAATTACAGCTTTATTATTACTTTTATCTTTACCAGTATTAGCTGGAGCT ATTACCATATTATTAACAGATCGAAATTTAAATACATCATTTTTTGACCCAGCTGGAGGGGGAGATCCATTTTATACC ACATTTTTTGATTTT
BLAST at The Wolbachia Project   BLAST at NCBI
Summary The Drosophila was found to be postive for Wolbachia.
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