Chemical Ecology of Xyleborus glabratus and Implications for Monitoring and Management

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Chemicl Ecology of Xyleorus glrtus nd Implictions for Monitoring nd Mngement Pul E. Kendr, Wyne S. Montgomery, Jerome Niogret, Elen Q. Schnell, nd Nncy D. Epsky USDA ARS, Sutropicl Horticulture Reserch Sttion 1361 Old Cutler Rod, Mimi, FL emil: pul.kendr@rs.usd.gov ----------------------------------------------------------------------------------------------------------------------- Conference on Lurel Wilt Disese nd Nturl Ecosystems: Impcts, Mitigtion, nd the Future 16-18 June 215

Applied Approch Trget = Adult femle RAB, host seeking flight Dispersl flight is risky Hrsh environment (dessiction, wind, rin) Predtion (e.g. drgonflies) No food ville Strong selection Relile cues to find host trees quickly Optimize timing of flight to minimize risks

Chemicl Ecology of RAB Initil reserch on RAB ttrctnts (Hnul et l. 28, Hnul & Sullivn 28) No evidence of pheromones (ggregtion or sex) No long rnge ttrction to fungl symionts No ttrction to ethnol (stndrd lure for mrosi eetles) Conclusions: RAB cn ttck helthy, unstressed trees Host tree voltiles = primry ttrctnts α copene, clmenene re likely ttrctnts from redy; ut cost/vilility mke them imprcticl for field lures Mnuk nd phoee oils identified s ttrctive its for RAB

ARS Mimi,FL Reserch RAB ttrctnts (29 pres.) Multidisciplinry pproch Field tests (forest sites) L tests (Archold Biol. Sttion, Lke Plcid) Behviorl iossys (ttrction, oring preferences) Electrophysiology techniques to mesure olfctory responses Chemicl smpling &nlysis (Mimi) Voltile collections Gs chromtogrphymss spectrometry (GC MS)

1. Method for Cpture of Live RAB Generl Method Lte fternoon (~4: pm) Ly cotton sheet on ground Bit = Freshly cut host wood Lure in mrosi eetles Collect with soft rush Kendr et l. 212 FL Entomol.

Unique Oservtion: Temporl Seprtion of Beetle Species Multiple species ttrcted to wood voltiles X. glrtus flies erlier (27 collection dtes, Apr Oct 211) Useful method for otining RAB in host seeking ehvior, the perfect stge to evlute ttrctnts L iossys Electrophysiology studies Numer of Femles Collected Numer of Femles Collected Numer of Femles Collected 14 12 1 8 6 4 2 16 163 17 173 18 183 19 193 2 7 6 5 4 3 2 1 6 5 sunset X. glrtus X. ffinis 16 163 17 173 18 183 19 193 2 X. ferrugineus 4 3 2 1 16 163 17 173 18 183 19 193 2 Time Intervl Kendr et l. 212 Environ. Entomol.

2. Electrontennogrphy (EAG) Technique to mesure olfctory response of ntennl receptors (How well does n insect smell prticulr chemicl?) Mount ntenn etween 2 electrodes Plce under purified ir flow Deliver chemicl smple Mesure receptor potentil A B C EAG l t Archold Biol. Sttion Smple EAG recordings from single RAB ntenn

EAG Technique Mjor chllenge: smll size of RAB ntenn (.3.4 mm length) Used gold, 2 pronged electrode Electrode modified y ttching flexile gold wire to one prong Modified ntennl electrode Scnning electron microgrph of RAB ntenn

Vlidtion of New EAG Methods Comprtive studies with 3 species: X. glrtus, X. ffinis, nd X. ferrugineus Ethnol = stndrd reference chemicl EAG responses to test chemicls could then e EAG Response (-mv) 2.5 2. 1.5 1..5 Ethnol. expressed s percentge reltive to ethnol X. gl X. ffin X. ferr 1 1 1 A. Silky wood B. Phoee oil C. Mnuk oil 8 8 8 EAG Response (%) 6 4 6 4 6 4 2 2 2 X. gl X. ffin X. ferr X. gl X. ffin X. ferr X. gl X. ffin X. ferr Kendr et l. 212. Environ. Entomol.

3. Bolt Boring Biossy No choice tests to ssess RAB oring ehviors 1 gllon ucket 1 olt plus 1 15 femle RAB Recorded # RAB oring nd loction on olt fter 2, 4, 6, 8, 24 hr (Positive oring = hlf of the ody inserted) Replicted 5x Kendr et l. 213 FL Entomol.

Composite Results Biossys (Adpted from Kendr et l. 214 PLoS ONE) 1 Silky (99%) Swmpy/Redy (91%) Percentge of Femles Boring 8 6 4 2 Avocdo (8%) Cmphor tree (5%) Lncewood (44%) Live Ok (%) 5 1 15 2 25 Time (hr) >7% oring on cut end; trees most susceptile to ttck fter pruning or injury.

4. Comprtive Field Tests Replicted field tests with freshly cut wood olts Silky most ttrctive = positive control; cptures normlized Determined reltive ttrction of 9 species within Lurcee 1. Normlized Cptures.8.6.4.2 c c c c. Silky Cmphor. Cl. By Sssfrs Swmpy Redy Avocdo Lncewood Spiceush Live Ok Control Kendr et l. 214 PLoS ONE

5. Chemicl Anlysis Emissions of terpenoids from olts correlted with RAB cptures in field 4 sesquiterpenes Mjor α cueene (pek 2) α copene (pek 3) Minor α humulene (pek 6) clmenene (pek 8) Kendr et l. 214 PLoS ONE

Current Hypothesis Host Loction, Acceptnce Long rnge: RAB in flight ttrcted to the odor plume from host trees. Terpenoids: α copene, α cueene, α humulene, clmenene (Hnul & Sullivn 28, Kendr et l. 211, 214); euclyptol (Kuhns et l. 214); other monoterpenes (Mrtini et l. 215) Mid rnge: RAB uses visul cues to find hosts of pproprite dimeter (Myfield nd Brownie 213) Short rnge: RAB detects (1) terpenoid grdients to find est site for ttck (vocdo: α copene nd α cueene) (Niogret et l. 213), nd (2) fungl odors = food ttrctnts (Hulcr et l. 211, Kuhns et l. 214) On contct: RAB tste nd feel the wood to confirm suitility (euclyptol, Kuhns et l. 214) Summry: Host must smell, look, tste, nd feel right efore oring initited.

Development of Field Lures for RAB First lures = phoee oil, mnuk oil (Hnul & Sullivn 28) However, tests in FL (29 21) indicted tht mnuk lures not very effective 2. Beetles / trp / week 1.5 1..5 X. glrtus (men ± SE) 8-wk test Alchu Co., FL c c. Phoe Gut W.Ind Mex Mnuk Contr (Kendr et l. 211. J. Chem. Ecol.)

Follow up Tests Confirmed Results Beetles / trp / wk 25 2 15 1 Xyleorus glrtus (men + SE) 12-wk test Highlnds Co., FL Beetles / week 35 3 25 2 15 1 Xyleorus glrtus Weekly Cptures over Time Phoee Mnuk Control Freeze 5 5 Phoee Mnuk Control c 2 4 6 8 1 12 14 Week (Kendr et l. 212. J. Econ Entomol.) Phoee cught 6X more RAB Mnuk lost ttrction fter 3 wk Unfortuntely, phoee lures no longer ville, mnuk lures only option for RAB monitoring

Evlution of Other Essentil Oils Field tests in 212 compred 7 essentil oils for RAB ttrction: (Homemde lures 2 ml oil) Mnuk Phoee Cue Ginger root Angelic seed Te tree Vlenci ornge (Agriculturl Reserch 212)

12 1 X. glrtus (men + SE) 4-wk test (25 Apr - 23 My 212) Beetles / trp / week 8 6 4 c 2 c c Mn Cu Ph Gin Ang Te Or Blk Essentil oil (Kendr et l. 213. Am. J. Plnt Science) Cue oil identified s new RAB ttrctnt (from erries of tiled pepper Piper cue)

Field Tests 213 Collortion with Synergy Semiochemicls Corp. (BC, Cnd) New lure = distilled cue oil in ule lure (enriched in sesquiterpenes) Field tests to compre efficcy nd longevity of commercil lures: Cue ule lure Mnuk lure Phoee lure United control

Test 1 (Highlnds County, FL) 7 6 Xyleorus glrtus (men + SE) RAB / trp / wk 5 4 3 8-wk Field Test (5 reps) Archold Biol. Sttion (Lke Plcid, FL) (28 Fe - 25 Apr 213) 6 5 Xyleorus glrtus - Weekly cptures Cue Mnuk Control 2 4 1 c RAB / wk 3 cue mnuk lnk 2 1 (Kendr et l. 214. J. Pest Science) 1 2 3 4 5 6 7 8 9 1 Week Cue ule lure cptured 3 times more RAB thn mnuk

Test 2 (Gldes County, FL) 2.5 Beetles / trp / week 2. 1.5 1..5 Xyleorus glrtus (men + SE) 8-wk Field Test Fisheting Creek Stte Prk (8 Mr - 3 My 213). Cue ule Mnuk ptch Control (Kendr et l. 214. J. Pest Science) At very low popultion levels, cue ule lure cptured 6 times more RAB thn mnuk

Test 3 (Highlnds County, FL) X. glrtus weekly cptures Beetles / trp / wk 18 16 14 12 1 8 6 4 2 Xyleorus glrtus (men + SE) 12-wk Field Test Highlnds Hmmock Stte Prk c c Beetles / week 16 14 12 1 8 6 4 2 Cue Phoee Mnuk Blnk Cue Phoee Mnuk Control 1 2 3 4 5 6 7 8 9 1 11 12 Week Cue > phoee > mnuk Cue field life of 3 months (Kendr et l. 215. J. Econ. Entomol.)

Lure Emissions Mnuk: High initilly, expon. decy (lrge surfce re: volume rtio) Cue: Low initilly, slow, stedy relese (smller surfce re: volume rtio) Quntity ( g) 3 25 2 15 1 5 α-copene B Quntity ( g) 1.6 1.4 1.2 1..8.6.4.2. 3 4 5 6 7 8 1 2 3 4 5 6 7 8 Time (d) Time (d) Cue Mnuk (Kendr et l. 215. J. Econ. Entomol.)

Cue ule lure Conclusions More ttrctive thn the mnuk lure Hs field life of t lest 3 months, due to extended low relese of sesquiterpenes Best product currently ville for detection of RAB (nd less expensive thn the mnuk lure) However, cue lure contins complex mixture of terpenoids; specific ttrctive chemicls hd not een confirmed.

Further Improvement of RAB Lure (Collortion with Synergy) Gol = Elucidte the primry ttrctnts in cue oil. Used frctionl distilltion to seprte whole oil into 17 frctions (sed on chemicl oiling point) Frctions formulted s ule lures EAG nlyses 2 choice iossys Only 2 frctions were significntly ttrctive to RAB; oth were high in α copene nd α cueene Decided to focus on α copene first

New Lure Evlutions Two prototype lures prepred Copi oil (9% α copene, no cueene) Proprietry oil product (5% α copene) 12 wk field test Compre new lures to cue lure (1% α copene, 1% α cueene) Conclusions ( ) α copene = primry ttrctnt Incresed copene = improved lure X. glrtus / trp / wk X. glrtus / wk 16 14 12 1 8 6 4 2 18 16 14 12 1 8 6 4 2 A B 5% Cop Cue Copi Control 5% Cop Cue Copi Control 1 2 3 4 5 6 7 8 9 1 11 12 Week Kendr et l. J. Pest Sci. (in review) c

Sources of Funding THANK YOU! USDA Ntionl Plnt Disese Recovery System Florid Avocdo Administrtive Committee Recent Pulictions Kendr, P. E., W. S. Montgomery, J. Niogret, E. Q. Schnell, M. A. Deyrup, nd N. D. Epsky. 214. Evlution of seven essentil oils identifies cue oil s most effective ttrctnt for detection of Xyleorus glrtus (Coleopter: Curculionide: Scolytine). J. Pest Sci. 87: 681 689. Kendr, P. E., W. S. Montgomery, J. Niogret, G. E. Pruett, A. E. Myfield III, M. McKenzie, M. A. Deyrup, G. R. Buchn, R. C. Ploetz, nd N. D. Epsky. 214. North Americn Lurcee: Terpenoid emissions, reltive ttrction, nd oring preferences of redy mrosi eetle, Xyleorus glrtus (Coleopter: Curculionide: Scolytine). PLoS ONE 9 (7): e1286. Kendr, P. E., J. Niogret, W. S. Montgomery, M. A. Deyrup, nd N. D. Epsky. 215. Cue oil lures: Terpenoid emissions, trpping efficcy, nd longevity for ttrction of redy mrosi eetle (Coleopter: Curculionide: Scolytine). J. Econ. Entomol. 18: 35 361. Kendr, P. E., W. S. Montgomery, M. A. Deyrup, nd D. Wkrchuk. Improved lure for redy mrosi eetle developed y enrichment of α copene content. J. Pest Sci. (in review).