Team members Mirian Michereff Embrapa Maria C. Blassioli-Moraes Embrapa Haruna Braimah CSIR/Crops Research Institute Raúl A. Laumann Embrapa Christine Woodcock Rothamsted Research- UK Michael A. Birkett RothamstedResearch-UK John A. Pickett RothamstedResearch-UK Institutions Embrapa Recursos Genéticos e Biotecnologia CSIR/Crops Research Institute(Ghana) Rothamsted Research(UK)
bjectives The overall objective will be achieved in 5 action steps: 1. Identify attractants for S. mangiferae from weevils and host plants; 2. Confirm behavioural activity of identified attractants in the laboratory; 3. Confirm the activity of locally sourced, collected and extracted semiochemicals in the field; 4. Demonstrate and extend the technologies to farmers and extension staff through farmer field schools; 5. Produce material for guidance of farmers in controlling and detecting S. mangiferae populations.
World distribution of S. mangiferae = Stablished = Localized = Spread Localized for the first time in Brazil 216, Rio de Janeiro
Semiochemical interactions Alelochemicals Intespecific interactions Kairomones (-/ +) Pheromones (Intraespecific) Ex: sex/agreggation pheromone
Mangoes from Brazil and Ghana, insects from Ghana Air-entrainment volatiles - Laboratory Picture: Francisco G. Schmidt Air-entrainment volatiles - field Picture: Francisco G. Schmidt
Colony Establishment CSIR -Crops Research Institute, Kumasi - Ghana
Expected x Achieved Results
management of the mango stone weevil, Sternochetus pa 5 25 GC-FID Chemical profile - flower 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 pa 5 25 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 pa 5 25 11 12 14 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 pa 5 25 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 pa 5 25 1 1 1 1 1 2 2 2 3 5 3 5 3 5 3 5 8911 8 911 12 8 9 9 8 11 12 9 11 13 14 13 14 13 13 BIC UBA Palmer Tommy 14 Golden drop disease antracnose 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 1) β-myrcene, 2) 3-carene, 3) limonene, 4) (Z)-ocimene, 5) (E)-ocimene 6) α-terpinolene, 7) (E)-linalool-oxide, 8) methyl benzoate, 9) linalool, 1) nonanal, 11) DMNT, 12) allocimene, 13) ethyl benzoate, 14) methyl salicylate, 15) decanal, 16) 2-methoxy-methyl benzoate, 17) α-copaene, 17)α-gurjujene, 18)β-caryophyllene, 19)α-humulene, 2) gemarcrene D, 21)β-selinene, 22) (E,E)-α-farnesene. 17 18 17 17 18 18 18 19 22 192 22 192 22 21 192 2 22 22 min min min min min
management of the mango stone weevil, Sternochetus GC-FID chemical profile of volatiles- Fruits pa 5 Bico 25 (green 2- cm) 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 min 6 Tommy (ripe- Market) pa 5 1 18 25 2 3 4 5 8 14 7 91 11 16 1 15 17 9 2 19 22 2425 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 Tommy (green market) pa 5 25 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 UBA (green 2 cm) pa 5 25 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 Golden drop with antracnose disease (green 2 cm) pa 5 25 1 12.5 15 17.5 2 22.5 25 27.5 3 32.5 1) α-pinene, 2) camphene, 3) β-pinene, 4) myrcene, 5) α-phellandrene, 6)3-carene, 7) α-terpinene, 8) limonene, 9)(E)-ocimene, 1) γ-terpinene, 11) terpinolene, 12) ethyl benzoate, 13) methyl salicylate, 14) elixene, 15) α-copaene, 16) sesquiterpene, 17) α- gurjunene, 18)β-caryophyllene, 19)β-cubebene, 19) γ-muurolene, 2) α-humulene, 21) β-gurjunene, 22) gemarcrened, 23) (E,E)- α-farnesene, 24) δ-cadinene. 23 min min min min
management of the mango stone weevil, Sternochetus Plant volatiles for weevil management 24 major compounds were identified from flower and fruit from GC - MS trace. Which compounds play a role on insect behavior? And the minor compounds? Are they important?
management of the mango stone weevil, Sternochetus Sensorial response to fruit VCs myrcene 5 H γ-butyrolactone 6-metil-5-hepten-2-one 2-methyl-methyl-butanoate furfural 1 2 3 4
management of the mango stone weevil, Sternochetus S.Mangiferaeantenna GC-EAD response Flower air-entrainment extracts EAG myrcene H furfural 6-metil-5-hepten-2-one 2-methyl-methyl-butanoate GC FiguraFlower. Female antennae response to floral mango volatiles. (name of compound/ri) 1) 2-methyl-methyl butanoate,/766, 5) furfural/799, 6) benzaldehyde/925, 7) 6-metil-5-hepten-2-one/965, 8) myrcene/982, 9) limonene/123, 11) benzyl isocyanide/151, 12) 2-methoxyphenol (guaicol)/159, 13) methyl benzoate /162, 14) unknown/175, 15) linalool/183, 18) ethyl benzoate/1143, 2) methyl salicylate/1168, 21) methyll-2-methoxybenzoate /1294. IR - DB-1
management of the mango stone weevil, Sternochetus S.Mangiferaeantenna GC-EAD response Fruit air-entrainment extracts myrcene 6-metil-5-hepten-2-one γ-butyrolactone 2-methyl-methyl-benzoate. Female antennae response to fruit mango volatiles. (Name of compound/ri) (1) 2-methyl-methyl butanoate /766, 2) γ-butyrolactone/848, 3) (E)-2- hexen-1-ol/853, 4) anisole/893, 6) benzaldehyde/925, 7) 6-metil-5-hepten-2-one/962, 8) myrcene/982, 1) (Z)-ocimene/12, 12) methyl bezoate/162, 15) linalool /183, 16) DMNT/115, 18) ethyl benzoate/1143, 19) naphtalene/1169, 2) methyl salicylate/1168, 21) metil-2-methoxybenzoate /1294).
management of the mango stone weevil, Sternochetus S.Mangiferaeantenna GC-EAD response Synthetic solution- Mango volatiles (flower and fruit) 1 2 EAG 2-methyl-methyl-butanoate 3 4 γ-butyrolactone 5 benzaldehyde 6 6-metil-5-hepten-2-one (Z)-ocimene 1 2 3 4 5 7 8 9 myrcene 8 6 H 7 linalool H GC guaiacol 9 H methyl salicylate
Noite Bioassays with synthetic solution Females Hexane Mix synthetic with racemic linalool Hexane Hexane * Mix synthetic with (S)-linalool Mix synthetic with (R)-linalool Hexane Males Mix synthetic with racemic linalool Hexane Mix synthetic with (S)-linalool Hexane Mix synthetic with (R)-linalool 1..8.6.4.2..2.4.6.8 1. Mix Fruit: butyrolactone, benzaldehyde, 6-methyl-5-hepten-2-one, myrcene, (Z)-ocimene, 2-methoxyphenol, linalool, methyl-salicylate, methyl-benzoate.
Absolute configuration of linalool produced by mango flowes H H (R) (S) 1 5 1 5 1 5 Co-injection flower + linalool 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 S Flower 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 R S Linalool racemic 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 1 5 R R-Linalool 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 Retention time(min)
Absolute configuration of linalool produced by mango fruits H H (R) (S) 1 5 1 5 R S 27. 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 S Co-injection fruit + linalool Fruit 27. 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 1 5 1 5 R S Linalool racemic 27. 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 R R-Linalool 27. 27.5 28. 28.5 29. 29.5 3. 3.5 31. 31.5 32. 32.5 Retention time (min)
Pheromone for mango weevil management FIVE specific Sternochetus mangiferae male compounds were indetified from GC-FID profile
GC-FID profile from air-entrainment extract from males and females of Sternochetus mangiferae
Behaviouralbioassays with live male and female odours Double choice olfactometer dour 1 dour 2 Release area
lfactometer bioassays with live insects Female responses Photophase Ar * Male p=.2 (44) Scotophase Male Female p=.65 (57) Ar * Female p=.3 (8) Ar ** Male p=.7 (25) 6, 4, 2,, 2, 4, 6, Number of females responding
lfactometer bioassays with live insects Male responses Photophase Ar Scatophase p=.12 Female (49) Male p=.44 Female (72) Ar Male p=.68 (76) Ar Female p=.17 (57) 4, 3, 2, 1,, 1, 2, 3, 4, Number of males responding
Nine VCS from host plants were identified with potential for attraction of mango weevil 2-methyl-methyl butanoate, γ-butyrolactony, benzaldehyde, 6-metil-5-hepten-2- one, myrcene, (Z)-ocimene, 2- methoxyphenol, linalool, methyl salicylate, These VCs appear to have a key role on the weevil attraction to host plants. The chemical analysis of extracts by GC-FID and the bioassays conducted suggest hat males are the sex pheromone producer. The insects present activity only during the scotophase
August/217 March/218 Next Steps Field experiment in Ghana Africa. Behavioural assays with insects detected in Brazil. Field experiment in Rio de Janeiro. Air-entrainment volatile collection with Brazilian weevils detected in Rio de Janeiro.
Lessons Learned and Bottlenecks Animal and Plant Health Inspection Service - Plant Protection and Quarantine APHIS - PPQ Were larva found feeding on the fruit, or only adults? Have additional detections occurred at this or other locations? Are pheromone or other types of traps available? Is trapping taking place around the detection site (a delimiting survey)? Has MAPA removed the tree and established a quarantine or additional phytosanitary requirements such as fruit surveys or fruit cutting around the detection site? Is trapping occurring in the commercial production areas in the San Francisco Valley and Natal, which do export to the United States? Is MAPA implementing additional phytosanitary precautious at the commercial packing houses? Is trapping occurring in the commercial production areas in states of Minas Gerais and Sao Paulo, which do not export to the United States?
Lessons Learned and Bottlenecks Quarantine Insects Not allowed to work freely with the insects, Univoltine insect - species that have just one brood per year, No colony established in laboratory, Lack of information regarding their bioecology, Infrastructure in the local Research Unit of the pest occurrence, Transport authorization of Sternochetus mangiferae for research purposes, The fact that Cenargen has a quarantine laboratory seems do not facilitate this process. It seems to us that this process will be very time consuming and will delay the research.
Acknowledges Dr. Tadeu Graciolli Guimarães - Embrapa Cerrados Team Mirian Michereff pós-doutorado Maria C. Blassioli-Moraes pesquisadora Haruna Braimah CSIR/Crops Research Institute (Ghana) Raúl A. Laumann pesquisador Christine Woodcock Rothamsted (UK) Michael A. Birkett Rothamsted (UK) John A. Pickett Rothamsted (UK) Haruna Braimah co-leader
MANG THE QUEEN F TRPICAL FRUITS 6 Thank you brigado miguel.borges@embrapa.br