Download gut microbiota mediate caffeine detoxification in the primary insect

GUT MICROBIOTA MEDIATE
CAFFEINE DETOXIFICATION IN THE
PRIMARY INSECT PEST OF COFFEE
JAVIER A. CEJA-NAVARRO, FERNANDO E. VEGA, ULAS KARAOZ1, ZHAO HAO1, STEFAN JENKINS, HSIAO CHIEN LIM1,
PETR KOSINA, FRANCISCO INFANTE, TRENT R. NORTHEN & EOIN L. BRODIE (2015)
SEMINAR MSC. BIOTECHNOLOGY - 13.01.2016
ESTHER HUBER
SUPERVISOR: DIPL.-BIOL. MIRJAM SELZER
CONTENT
 Coffee – Coffea
 Caffeine
 The coffee berry borer - Hypothenemus hampei
 Understanding of metabolism and ecology of the pest
 Caffeine reduction in the insect gut of H. hampei
 Caffeine-degrading bacteria in the gut of H. hampei
 Oxygen profiles in the gut of the coffee berry borer
 Conclusion
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COFFEE – COFFEA
 Coffee (Coffea) - Plant genus of the family Rubiaceae
 Currently 124 known species
 Commercially important species:

Coffee arabica (Coffea arabica)
Coffee robusta (Coffea canephora)
 Contains caffeine as a toxic alkaloid
 Content of caffeine:

Coffea arabica  1% Seed-DW
Coffea canephora  1,7-2,4% Seed-DW
SEMINAR MSC. BIOTECHNOLOGY - ESTHER HUBER
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CAFFEINE
 Bitter taste
 negative effects on insects, arachnids, slugs and snails:
 Paralyzes and intoxicated
 Negative effects on DNA repair
and recombination pathways
 delay of cell cycle progression
  natural protection against herbivory
https://upload.wikimedia.org/wikipedia/commons/8/83/FruitColors.jpg
https://de.jura.com/-mages/barista%20tips/barista%20kaffeebohne.jpg?la=de&mw=305
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THE COFFEE BERRY BORER –
HYPOTHENEMUS HAMPEI
 Body length up to 2 mm, while typical body shape of the bark beetle
 Most common pest of coffee plants
 Does not cause damage to leaves, branches or trunks
 Infects immature coffee berries
http://www.cpt.com.br/imagens/enviadas/materias/materia10058/broca-cafeeiro-superadensado-cursos-cpt.jpg
SEMINAR MSC. BIOTECHNOLOGY - ESTHER HUBER
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THE COFFEE BERRY BORER –
HYPOTHENEMUS HAMPEI
 Build galleries within the coffee seed, where females deposits 31-119 eggs
 At 27 °C larval stages in 15 days, the pupal stage will go through in 7 days
 Total life cycle (28 - 34 days) on / in green coffee beans
 Yearly losses in Brazil $285–315 million
http://www.cpt.com.br/imagens/enviadas/materias/materia10058/broca-cafeeiro-superadensado-cursos-cpt.jpg
SEMINAR MSC. BIOTECHNOLOGY - ESTHER HUBER
http://www.princessradhafarm.com/images/cbbpic.jpg
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THE COFFEE BERRY BORER –
HYPOTHENEMUS HAMPEI
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UNDERSTANDING OF METABOLISM
AND ECOLOGY OF THE PEST
 horizontal gene transfer from bacteria on H. hampei:
HhMAN1 – gene of the digestive enzyme Mannanase (Acuña et al., 2012)
  enabled the insect to exploit the primary carbohydrate reserve in coffee beans
 no correlation between caffeine concentrations in the beans of
12 Coffea species and resistance to H. hampei
  suggesting that insect possesses mechanisms to degrade or
efficiently excrete caffeine
  hypothesis that caffeine degradation in H. hampei is mediated by the activity
of its gut microbiota
SEMINAR MSC. BIOTECHNOLOGY - ESTHER HUBER
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CAFFEINE REDUCTION
IN THE INSECT GUT OF H. HAMPEI
 Examination of colonies from 7 coffee growing areas worldwide, 1 laboratory
colony and 3 caffeine-sensitive bark beetle species
 Minimal diet:
 Comparative study:
Green beans of C. arabica
Minimal diet and 3 broad-spectrum antibiotics
 Determined by incorporating fluorescent sphere tracers and FTIR- spectroscopy,
respectively GC-MS
 Measurement of:
 Quantity
of diet consumed
 Residence time in digestive tract
 Amount excreted caffeine
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CAFFEINE REDUCTION
IN THE INSECT GUT OF H. HAMPEI
SEMINAR MSC. BIOTECHNOLOGY - ESTHER HUBER
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CAFFEINE-DEGRADING BACTERIA
IN THE GUT OF H. HAMPEI
 Biogeographical analysis
 Identification of relevant bacteria
 16S rRNA Sequenzanalyse
 Screening for the ndmA-gene
(α-subunit of a caffeine demethylase)
  Pseudomonas fulva possesses the gene
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CAFFEINE REDUCTION
IN THE INSECT GUT OF H. HAMPEI
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OXYGEN PROFILES IN THE GUT OF
THE COFFEE BERRY BORER
 Caffeine demethylase is an oxygen-dependent monooxygenase
 Measurement using microelectrodes
 Intestine contains radial oxygen gradient
of microaerophilic to anaerobic conditions
  Place for aerobic processes:
transformation of caffeine
  Place for anaerobic processes:
fermentation of sugars of coffee beans
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CONCLUSION
 Only insect that specializes in living
in / on coffee berries
 Treatment with antibiotics restricts Fitness
 Inoculation of caffeine-degrading gut microbiota is possible –
but Insects did not recover their ability to reproduce after inoculation!
 Furthermore, to investigate:
  Determining the minimal set of microorganisms to reinstate reproductive fitness
 effect of coffee species with varying caffeine concentrations on the microbiome
 Research provides novel avenues to develop biocontrol strategies
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ANY QUESTIONS ?
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LITERATURE

Javier A. Ceja-Navarro, Fernando E. Vega, Ulas Karaoz, Zhao Hao, Stefan Jenkins, Hsiao
Chien Lim, Petr Kosina, Francisco Infante, Trent R. Northen & Eoin L. Brodie (2015): Gut
microbiota mediate caffeine detoxification in the primary insect pest of coffee. NATURE
COMMUNICATIONS, 6:7618

Pflanzenforschung (Bundesministerium für Bildung und Forschung): Bakteriengen macht
Insekten zu Schädlingen; http://www.pflanzenforschung.de/index.php?cID=5537

Ricardo Acuña, Beatriz E. Padilla, Claudia P. Flórez-Ramos, José D. Rubio, Juan C. Herrera,
Pablo Benavides, Sang-Jik Lee, Trevor H. Yeats, Ashley N. Egan, Jeffrey J. Doyle, Jocelyn K. C.
Rose (2012): Adaptive horizontal transfer of a bacterial gene to an invasive insect pest of
coffee. Proceedings of the National Academy of Sciences USA vol. 109 no. 11: 4197–4202.

Gokulakrishnan, S., Chandraraj, K. & Gummadi, S. (2005): Microbial and enzymatic methods
for the removal of caffeine. Enzyme Microb. Technol. 37, 225–232.
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