AUTHOR=Vega Fernando E. , Emche Sarah , Shao Jonathan , Simpkins Ann , Summers Ryan M. , Mock Meredith B. , Ebert Dieter , Infante Francisco , Aoki Sayaka , Maul Jude E. 

TITLE=Cultivation and Genome Sequencing of Bacteria Isolated From the Coffee Berry Borer (Hypothenemus hampei), With Emphasis on the Role of Caffeine Degradation

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.644768

DOI=10.3389/fmicb.2021.644768

ISSN=1664-302X

ABSTRACT=<p>The coffee berry borer, the most economically important insect pest of coffee worldwide, is the only insect capable of feeding and reproducing solely on the coffee seed, a food source containing the purine alkaloid caffeine. Twenty-one bacterial species associated with coffee berry borers from Hawai’i, Mexico, or a laboratory colony in Maryland (<italic>Acinetobacter</italic> sp. S40, S54, S55, <italic>Bacillus aryabhattai</italic>, <italic>Delftia lacustris</italic>, <italic>Erwinia</italic> sp. S38, S43, S63, <italic>Klebsiella oxytoca</italic>, <italic>Ochrobactrum</italic> sp. S45, S46, <italic>Pantoea</italic> sp. S61, <italic>Pseudomonas aeruginosa</italic>, <italic>P. parafulva</italic>, and <italic>Pseudomonas</italic> sp. S30, S31, S32, S37, S44, S60, S75) were found to have at least one of five caffeine N-demethylation genes (<italic>ndmA</italic>, <italic>ndmB</italic>, <italic>ndmC</italic>, <italic>ndmD</italic>, <italic>ndmE</italic>), with <italic>Pseudomonas</italic> spp. S31, S32, S37, S60 and <italic>P</italic>. <italic>parafulva</italic> having the full complement of these genes. Some of the bacteria carrying the ndm genes were detected in eggs, suggesting possible vertical transmission, while presence of caffeine-degrading bacteria in frass, e.g., <italic>P. parafulva</italic> (<italic>ndmABCDE</italic>) and <italic>Bacillus aryabhattai</italic> (<italic>ndmA</italic>) could result in horizontal transmission to all insect life stages. Thirty-five bacterial species associated with the insect (<italic>Acinetobacter</italic> sp. S40, S54, S55, <italic>B. aryabhattai</italic>, <italic>B. cereus</italic> group, <italic>Bacillus</italic> sp. S29, S70, S71, S72, S73, <italic>D. lacustris</italic>, <italic>Erwinia</italic> sp. S38, S43, S59, S63, <italic>K. oxytoca</italic>, <italic>Kosakonia cowanii</italic>, <italic>Ochrobactrum</italic> sp. S45, S46, <italic>Paenibacillus</italic> sp. S28, <italic>Pantoea</italic> sp. S61, S62, <italic>P. aeruginosa</italic>, <italic>P. parafulva</italic>, <italic>Pseudomonas</italic> sp. S30, S31, S32, S37, S44, S60, S75, <italic>Stenotrophomonas</italic> sp. S39, S41, S48, S49) might contribute to caffeine breakdown using the C-8 oxidation pathway, based on presence of genes required for this pathway. It is possible that caffeine-degrading bacteria associated with the coffee berry borer originated as epiphytes and endophytes in the coffee plant microbiota.</p>