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Composition and method for attraction of emerald ash borer agrilus planipennis fairmaire (coleoptera: buprestidae)

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Composition and method for attraction of emerald ash borer agrilus planipennis fairmaire (coleoptera: buprestidae)


The invention disclosed relates to a novel composition and use thereof, for the attraction and detection of emerald ash borer, Agrilus planipennis Fairmaire. The composition comprises (3Z)-dodecen-12-olide and ash foliar or cortical volatiles (green leaf volatiles) associated with a prism trap of a color in the green range of the visible light spectrum. A significant increase in the capture of male A. planipennis is achieved when traps were deployed in the upper tree canopy. This invention is the first demonstration of increased attraction with a combination of a pheromone and a green leaf volatile in a Buprestid species.
Related Terms: Cortical Emerald Pheromone Demon Prism Tiles

USPTO Applicaton #: #20130028858 - Class: 424 84 (USPTO) - 01/31/13 - Class 424 
Drug, Bio-affecting And Body Treating Compositions > Baits, Attractants, Or Lures (non-food)

Inventors: Peter J. Silk, David Magee, Krista Ryall, Peter Mayo

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The Patent Description & Claims data below is from USPTO Patent Application 20130028858, Composition and method for attraction of emerald ash borer agrilus planipennis fairmaire (coleoptera: buprestidae).

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This application claims the benefit of U.S. Provisional patent application 61/513,054 filed Jul. 29, 2011.

BACKGROUND OF THE INVENTION

The emerald ash borer, Agrilus planipennis Fairmaire, (Coleoptera: Buprestidae) is an invasive Palearctic species that has killed millions of ash trees (Fraxinus spp. L.) (Oleaceae) in the USA and Canada (Cappaert et al. 2005; Poland and McCullough 2006). Although initially detected near Detroit, Mich. in 2002, there is evidence that populations of this invasive species had been present in Michigan, USA and Ontario, Canada since the mid-1990s (Seigert et al. 2007). Since then, it has spread rapidly and has been detected in 15 states and two provinces, Ontario and Quebec, in Canada (EAB 2010). Movement of infested firewood and nursery stock has exacerbated its spread and large scale devastation of ash trees is predicted (Marchant 2006). Early detection of A. planipennis infestations has proven difficult because visual signs and symptoms, such as D-shape exit holes, epicormic shoots, bark deformities, and thinning crowns, usually appear only on heavily infested trees a year or more after populations have been established (Cappaert et al. 2005; de Groot et al. 2006, 2008; Poland and McCullough 2006). Development of a monitoring system is critical for early detection of A. planipennis populations, which would aid in management and control decisions. In order to maximize detection efficacy, a better understanding of the behavior and chemical ecology of adult A. planipennis is needed.

Adult A. planipennis are typically active between 0600-1700 h, particularly when the weather is warm and sunny (Yu 1992; Rodriguez-Saona et al. 2007), with mating occurring from 0900-1500 h and lasting for 20-90 min. Yu (1992) observed that adults preferred trees in open areas with direct sunlight and that during rainy or cloudy weather they tended to rest in cracks in the bark or on the foliage. Adult beetles, particularly males, spend much of their time in the canopy feeding and flying short distances (Lance et al. 2007; Lelito et al. 2007; Rodriguez-Saona et al. 2007). Indeed, traps in the mid-upper ash canopy capture more adults than traps hung below the canopy (Lance et al. 2007; Francese et al. 2007, 2008; Crook et al. 2008, 2009) and traps in locations exposed to direct sunlight (i.e. on the edge or near a gap) generally catch more adults than those in shaded locations (Poland et al. 2005; McCullough et al. 2006, 2009; Francese et al. 2008; Lyons et al. 2009).

Crook and Mastro (2010) reviewed the considerable progress made towards developing a trap that is effective at capturing A. planipennis (Francese et al. 2005, 2007, 2008, 2010; Crook et al. 2008, 2009; Lelito et al. 2007, 2008; McCullough et al. 2008). Color has been identified as an important factor affecting trap captures, with purple shown to be highly attractive (Francese et al. 2005, 2008; Crook et al. 2008). Purple traps typically catch more females than males (Francese et al. 2008; Crook et al. 2009), due to A. planipennis response to light in both the blue and red range of the visible spectrum (Crook et al. 2009). Currently, a sticky purple prism trap is utilized in surveys for A. planipennis in the United States (Francese et al. 2008; Crook and Mastro 2010). Adult A. planipennis also respond to green light in the 540-560 nm range of wave length 540-560 nm (Crook et al. 2009), with green traps capturing two to three times as many adults as purple traps. Crook et al. 2009 also found that dark green (24% reflectance) and light green (64% reflectance) caught more beetles than purple. Also, Francese et al (2010b) Can. Entomol. 142: 596-600 tested purple vs light green (540 nm, 64% reflectance) traps and reported that green caught more EAB, particularly males. Also, Francese et al. (2010a) J Econ Ent 103: 1235-1241 studied different green wavelengths and different reflectances, and concluded that the best trap would be a green trap with a wave length of 540 nm and 49% reflectance. Green traps typically have a bias towards males in trap captures (Lance et al. 2007; Rodriguez-Saona et al. 2007; Lelito et al. 2008; Crook et al. 2009). However, green traps typically catch more adults only when deployed high in the tree canopy. Thus, trap deployment, as well as color and lure combination, must be considered when evaluating traps for a monitoring program, as trap captures are likely influenced by adult preferences and behavioral activity patterns.

Numerous studies have described the chemical ecology of A. planipennis (Crook and Mastro 2010) and two types of host volatiles have been demonstrated to be attractive: bark sesquiterpenes (Poland and McCullough 2006; Crook et al. 2008) and green leaf volatiles (Poland et al. 2004, 2005, 2006, 2007; Rodriguez-Saona et al. 2006; de Groot et al. 2008; Grant et al. 2010). Girdled or stressed ash (Poland and McCullough 2006; Crook et al. 2008) are attractive to both sexes, as are Manuka and Phoebe oils which contain, in part, the sesquiterpenes emitted by stressed Fraxinus spp. (Crook et al. 2008; Crook and Mastro 2010; Grant et al. 2010). Of the green leaf volatiles, one compound in particular, (3Z)-hexenol, is highly antennally active and attractive to males (de Groot et al. 2008; Grant et al 2010). These results indicate that specific host volatiles act as kairomones in the chemical ecology of A. planipennis and these compounds may provide useful detection tools.

Much of the literature on the mating behavior of buprestids (e.g. Rodriguez-Saona et al. 2006; Lelito et al. 2007; Akers and Nielsen 1992; Gwynne and Rentz 1983; Carlson and Knight 1969) has described the use of visual and tactile cues for mate location. For buprestids, including those in the genus Agrilus, host location has been described as occurring first by olfactory processes and then mate location by visual, or by vibratory and/or tactile cues (Carlson and Knight 1969). However, Dunn and Potter (1988) showed attraction of A. bilineatus (Weber) males to cages containing females compared to host-logs only, suggesting the use of a female-produced pheromone.

Limited progress has been made into the pheromone chemistry of A. planipennis. Previous work suggested the presence of a contact pheromone (Lelito et al. 2007), subsequently identified by our research group as 9-methylpentacosane, which appears only on the cuticle of female A. planipennis at sexual maturity (7-10 d old) and stimulates full copulatory activity in males upon antennal contact (Silk et al. 2009), although 3-methyltricosane may also be involved as an additional component (Lelito et al. 2009). Bartelt et al. (2007) identified a volatile, antennally-active predominantly female-produced macrocyclic lactone, (3Z)-dodecen-12-olide [(3Z)-lactone], which was the first putative volatile pheromone described for A. planipennis, but no behavioral activity was reported.

Pureswaran and Poland (2009) reported that males were able to locate and identify females at close range using olfaction and an unidentified volatile cue. Here, we use GC-EAD in combination with field trapping and olfactometry to test whether (3Z)-lactone elicits behavioral responses in A. planipennis either alone or in combination with host kairomones (bark sesquiterpenes or green leaf volatiles). We tested various lure combinations on both purple and green traps, as both colors have been shown to be attractive. We also tested the lactone stereoisomer, (3E)-lactone, for its effect on A. planipennis behavior because preliminary studies suggested that exposure to UV-light catalyzes the isomerization of (3Z) to the (3E)-lactone and A. planipennis adults are known to favor sunny locations.

SUMMARY

OF THE INVENTION

This invention provides the first behavioral evidence for a volatile pheromone of A. planipennis in combination with host foliar volatiles in association with a trap of a color in the green range of the visible light spectrum, contributing to the knowledge of the chemical ecology and the development of improved tools for the detection of A. planipennis infestations.

According to one aspect of the present invention, a composition for the attraction of A. planipennis is provided, comprising (a) (3Z)-dodecen-12-olide and (b) ash foliar volatiles, associated with a trap of a color in the green range of the visible light spectrum.

According to one embodiment of the composition aspect of the invention, the amount of (3Z)-dodecen-12-olide is a source dosage which emits 2.4-160 μg of (3Z)-dodecen-12-olide per day at about 25° C.

According to another embodiment of the composition aspect of the invention, the ash foliar volatiles comprise (3Z)-hexenol.

According to yet another embodiment of the composition aspect of the invention, the ash foliar volatiles comprise (3Z)-hexenol of a source dosage which emits 50-100 mg per day at about 25° C.

According to yet another embodiment of the composition aspect of the invention, the trap is a sticky prism tap of a green color defined by a wave length of 540-560 nm and a reflectance of 24-64% e.g. 540 nm and 49% reflectance.

According to yet another embodiment of the invention, the amount of (3Z)-dodecen-12-olide is a source dosage which emits 2.4-22 μg per day of (3Z)-dodecen-12-olide and the amount of (3Z)-hexenol is a source dosage which emits 40-60 mg per day of (3Z)-hexenol.

According to another aspect of the present invention, a method for the attraction of A. planipennis is provided, comprising applying to an insect habitat an insect attracting amount of (3Z)-dodecen-12-olide and ash foliar volatiles, associated with a trap of a color in the green range of the visible light spectrum.

According to one embodiment of the method aspect of the invention, the amount of (3Z)-dodecen-12-olide is a source dosage which emits 2.4-160 μg of (3Z)-dodecen-12-olide per day at about 25° C.

According to another embodiment of the method aspect of the invention, the ash foliar volatiles comprise (3Z)-hexenol.

According to yet another embodiment of the method aspect of the invention, the ash foliar volatiles comprise (3Z)-hexenol of a source dosage which emits 50-100 mg per day at about 25° C.

According to yet another embodiment of the method aspect of the invention, the trap is a sticky prism tap of a green color defined by a wave length of 540-560 nm and a reflectance of 24-64% e.g. wavelength 540 nm and 49% reflectance.

According to yet another aspect of the invention, a kit for the attraction of A. planipennis is provided, wherein the components (a) and (b) of the composition are maintained separately until use, and associated for use with a trap of a color in the green range of the visible light spectrum. The kit comprises a septum containing an insect attracting amount of (3Z)-dodecen-12-olide dissolved in a volatile solvent e.g. hexane which evaporates before use, leaving the (3Z)-dodecen-12-olide to emit therefrom, and a bubble cap containing an effective amount of neat ash volatiles e.g. (3Z)-hexenol, associated with the trap and maintained separately until use.

According to one embodiment of the kit aspect of the invention, the amount of (3Z)-dodecen-12-olide is a source dosage which emits 2.4-160 μg of (3Z)-dodecen-12-olide per day, and the ash foliar volatiles comprise (3Z)-hexenol of a source dosage which emits 50-100 mg per day, at about 25° C., and wherein the trap is a prism sticky trap of a green color defined by a wave length of about 540-560 nm and a reflectance of 24-64%.

When a sticky trap is used, no insecticide is required. However, if a non-sticky trap is used, an appropriate insecticide is included.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a flow diagram illustrating a process according to the invention for the synthesis of (3Z)-dodecen-12-olide.

FIG. 2 is a flow diagram illustrating a process according to the invention for the synthesis of (3E)-dodecen-12-olide.

FIG. 3 is a graph illustrating electroantennographic (EAG) dose-response curves of male and female A. planipennis antennae to (3Z)-dodece-12-olide (3ZLac) and (3E)-dodece-12-olide (3ELac) according to dosages applied to stimuli cartridges. EAG dose-responses (mean±SEM) are presented relative to a positive control standard (geranyl acetone, 1 μg applied dose).

FIG. 4 illustrates GC-FID/EAD responses of male and female A. planipennis antennae. The FID trace is a synthetic mixture of (3E)- and (3Z)-lactones.

FIGS. 5a-d are graphs illustrating proportions of male A. planipennis crawling up the test vs. control arms of a Y-tube olfactometer in 12 independent trials in response to: (a) (3E)-lactone, (3Z)-lactone or a 60:40 combination; (b) Phoebe oil (25 μl) alone or combined with either (3E)-lactone or (3Z)-lactone); (c) Phoebe oil (2.5 μl) alone or combined with (3E)-lactone or (3Z)-lactone; and (d) (3Z)-hexenol alone or combined with either (3E)-lactone or (3Z)-lactone. For each stimulus, the test treatment was compared with the control using a chi-square goodness of fit test.

FIG. 6 are bar graphs illustrating mean (±SE) catches of male and female A. planipennis on purple sticky prism traps baited with various combinations of (3Z)- and/or (3E)-lactone and host volatiles in field experiments carried out at two sites in (a) 2008 and two sites (b, c) in 2009. Sites were analyzed separately in 2009 due to the differences in sex ratio. Note differences in scale of X-axis. Prior to analyses, data were transformed using a natural log (n+1), however untransformed data are presented. Error bars reflect + or − one standard error of the least squares means. In 2008 (FIG. 2a), (3E)-lactone was not tested except that it was present in the synthetic (3Z)-lactone at 2%.

FIG. 7 are bar graphs illustrating mean (±SE) catches of male and female A. planipennis on green sticky prism traps baited with the different attractant combinations at (a) Anika and McKellar sites combined and (b) sites in Michigan, USA. Plotted values reflect the least squares means of 12 replicate blocks in total (untransformed data). Statistics (P>F) apply to natural log (n+1)-transformed data following ANOVA. Error bars reflect + or − one standard error of the least squares means.

FIG. 8. Dose-response curve for male and female A. planipennis to increasing dose of (3Z)-lactone loaded on rubber septa and deployed on dark green sticky traps in combination with (3Z)-hexenol. Plotted values reflect the means of 10 replicate blocks in total (untransformed data). Statistics (p>F) apply to natural log (n+1)-transformed data following ANOVA. Error bars reflect + or − one standard error of the mean. Letters above bars indicate significant differences among treatments compared against the control within each sex.

FIG. 9 (a and b). Influence of (3Z)-lactone with and without (3Z)-hexenol on attraction of male and female A. planipennis to dark green sticky traps placed at different heights with respect to the ash canopy: (a) low and (b) high. Plotted values reflect the least squares means of 10 replicate blocks in total (untransformed data). Statistics (p>F) apply to natural log (n+1)-transformed data following ANOVA. Error bars reflect + or − one standard error of the least squares means.

DETAILED DESCRIPTION

OF THE INVENTION Methods and Materials

Source of Insects.

Trees with larval A. planipennis were felled near Windsor and Sarnia, Ontario; infested logs were transported to the Great Lakes Forestry Centre in Sault Step Marie, Ontario. Storage and rearing protocols have been previously reported (Silk et al. 2009). Emerged adults were sexed and virgin males and females were kept on a 16:8 h L:D cycle and supplied with water and foliage of evergreen ash, Fraxinus uhdei (Wenzig) Linglesh.

Volatile Collection.

Volatiles were collected from two groups of virgin adult males (n=18 and n=8) and two groups of virgin adult females (n=17 and n=18) feeding on ash leaves in separate 250 ml glass chambers (16:8 L:D at 22° C.). Adults were 10 d old when placed in the chambers in groups of 6-8 at one time; and were replaced as they died over the volatile collection period. Filtered air was drawn from the chambers at ˜0.1 L/min onto a pre-conditioned Super-Q® filter (˜200 mg) for 10-11 d. Volatiles were eluted using methylene chloride (3×2 mL) and concentrated to 10-20 μl under dry nitrogen.



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stats Patent Info
Application #
US 20130028858 A1
Publish Date
01/31/2013
Document #
13559748
File Date
07/27/2012
USPTO Class
424 84
Other USPTO Classes
International Class
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Drawings
9


Cortical
Emerald
Pheromone
Demon
Prism
Tiles


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