Identification of aggregation pheromone of brown marmorated stink bug

by creating stereoisomeric libraries of 1-bisabolen-3-ols

Ashot Khrimian, Aijun Zhang, Donald C. Weber, Hsiao-Yung Ho, Jeffrey R. Aldrich, Karl E. Vermillion, Maxime A. Siegler, Shyam Shirali, Filadelfo Guzman, and Tracy C. Leskey USDA-ARS, Beltsville Maryland

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GC-MS total ion chromatograms Hsiao-Yung Ho

A and B are male-specific compounds not found in H. halys females

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H. halys male aeration

Synthetic murgantiols

Aggregation pheromone of harlequin bug, Murgantia histrionica

Breakthrough #1 Hsiao-Young Ho, 2009

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1,10-bisaboladien-3-ol total 8 stereoisomers

10,11-epoxy-1-bisabolen-3-ol total 16 stereoisomers

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Dictionary of Terpenoids, v.1, 1991

Natural occurrence of 1,10-bisaboladien-3-ols

and 10,11-epoxy-1-bisabolen-3-ols • Representative 1,10-bisaboladien-3-ol (a.k.a. zingiberenol)

was isolated from ginger, Zingiber officinale • 1,10-Bisaboladien-3-ols were identified as rice stalk sting bug,

Tibaca limbativebtris, pheromone • 10,11-Epoxy-1-bisabolen-3-ol (called murgantiol) was

identified as aggregation pheromone of harlequin bug, Murgantia histrionica

• Male brown marmorated stink bug, Halyomorpha halys, produce 10,11-epoxy-1-bisabolenols

• In all four studies, absolute configurations and field attractiveness of pheromones have not been demonstrated

Breakthrough #2

Two stereoisomers from one reaction

X-Ray (Cu Kα) structure determination of intermediate triol

Displacement ellipsoid plot of crystalline RSRS triol

Breakthrough #3 Maxime Siegler, JHU

Synthesis of individual stereoisomers of 10,11-epoxy-1-bisabolen-3-ol

(3S,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol, main component of

brown marmorated stink bug aggregation pheromone

Synthetic work: Summary

We synthesized and streochemically characterized previously unknown:

• All 8 possible stereoisomers of 1,10-bisaboladien-3-ol

• All 16 possible 1-bisabolen-3,10,11-triols • All 16 possible 10,11-epoxy-1-bisabolen-3-ols

H. halys male airborne extract

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H. halys male airborne extract + SSRS

BMSB main pheromone component (A) identified!

Breakthrough #4

Breakthrough #5

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BMSB minor pheromone component (B) identified!

Brown Marmorated Stink Bug Pheromone Identification: Summary

• Main component of BMSB aggregation pheromone (A) has been identified as (3S,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol

• Minor component of BMSB aggregation pheromone (B) has been identified as (3R,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol

• Availability of all 16 stereoisomers and combination of two chiral columns streamlined the assignments and bioassay confirmed the identification.

A B

Breakthrough #6 H. halys captures in pyramid traps with pheromone components

(Don Weber)

Treatment Adults Nymphs

#2 (M4 ) 159.0 ± 12.4 b 5.4 ± 2.5

#4 (control) 49.6 ± 14.3 b 6.2 ± 3.1

#6 (M4+Cs) 201.2 ± 23.8 b 7.8 ± 3.7

#9 (M4+Cs+220) 174.8 ± 45.9 b 6.8 ± 1.6

#10 (RM4+Rcis) 758.4 ± 164.1a 7.2 ± 1.8

Breakthrough #7 Trapping H. halys with mixed-isomer lures

(Tracy Leskey)