Progress 11/15/07 to 11/14/11
Outputs
OUTPUTS: Objective 1. Chemical aspects of host-finding strategies of Cerambycidae and Buprestidae. A. Calidiellum rufipennis (Coleoptera: Cerambycidae), is an invasive secondary pest of Cupressaceae. Aerations showed males release a volatile pheromone, (3R)-3-hydroxy-2-hexanone. Significantly more beetles of both sexes responded to field traps with pheromones. B. Agrilus anxius, (Coleoptera: Buprestidae) is a native which attacks stressed birches. Volatiles of bark samples of healthy and stressed grey birch (Betula populifolia) were collected. Unfortunately analysis was unsuccessful. Objective 2. Investigate mating behavior of Cerambycidae (C. rufipenne) and Buprestidae (A. anxius, A. bilineatus and A. planipennis). A. Male C. rufipenne respond to female C. rufipenne only upon contact with their cuticular hydrocarbons. We synthesized dimethyl-5,7-nonacosane, which is female-specific, and confirmed that it is the contact pheromone. This finding is published. We examined mating ecology of A. anxius, A. planipennis and A. bilineatus. Laboratory mating of all 3 shared a highly stereotyped pattern. Agrilus planipennis mated for 1 h. A. anxius and A. bilineatus mated for less then10 m. Mating pairs were separated at different times and dissected. In all species, sperm is transferred in a spermatophore at the end of copulation. Agrilus planipennis was unique in bundling sperm into packets within spermatophores. Sperm moves rapidly out of the spermatophore, which is gone by 3 hours post-mating. In A. planipennis and A. anxius, fecundity is increased by multiple mating. Given opportunity, both sexes mate daily. Individuals show no preference for previous mates or body size. Work on mating and sperm kinetics has been shared in posters and talks. Work on multiple mating and fecundity in A. planipennis is in press. A manuscript of the parallel study on A. anxius is in preparation. Objective 3. Survey for natural enemies of Cerambycidae and Buprestidae. A. Cerambycidae: We examined 10 logs infested with C. rufipennis and found 25% overwintering mortality (N=1023). We found 86 parasitoid cocoons. B. Buprestidae: We concentrated on pathogens and Cerceris fumipennis (Hymenoptera: Crabronidae). C. fumipennis provisions her nest with adults of Buprestidae. Since 2008, we have located 105 wasp colonies in CT. We trained a network of 29 volunteers who monitored 40 sites for A. planipennis over 2 summers. We collected > 2000 buprestids (14 genera, 32 species). Experiments showed C. fumipennis uses cuticular hydrocarbons to recognize prey. Results of C. fumipennis monitoring will be published this year. We published on anomalous C. fumipennis prey items. Data on C. fumipennis prey recognition was shared at conferences. We examined C. fumipennis-collected prey for pathogens. In both BBB and Dicera lurida (Buprestidae), we observed microsproridia which look like a newly described Cystosporogenes sp. infesting BBB. We have sequenced pathogenic nematodes from BBB and D. lurida. They share 96% of their sequence with a nematode Deladenus siricidicola, which infests woodwasp Sirex noctilio. Work on description and DNA sequencing is ongoing. PARTICIPANTS: PARTICIPANTS: Volatile pheromones of SJCB; Dr. Lawrence Hanks, UI Urbana-Champaign and Dr. Jocelyn Millar, UC Riverside. EAB mating and fecundity; Dr. Melody Keena, USFS at Hamden CT. Biosurveillance was conducted with the help of volunteer Wasp Watchers, including volunteers from White Memorial Conservation Center, Bartlett Arboretum, State Foresters, US FS, Master Gardners, and numerous garden club members and arborists. Host recognition of C. fumipennis; Dr. Peter Silk, Canadian Forest Service. Buprestid pathogens, Dr. Charles Vossbrinck, CAES. TARGET AUDIENCES: State and private foresters, nursery owners, green industry professionals, and home owners. TARGET AUDIENCES: TARGET AUDIENCES: State and private foresters, nursery owners, green industry professionals, and home owners. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The identity, (3R)-3-hydroxy-2-hexanone, and role, aggregation, of the male-produced pheromone of Callidiellum rufipennis was discovered. The identity, dimethyl-5,7-nonacosane of the contact sex pheromone of C. rufipennis was also discovered. We ascertained that Agrilus uses a spermatophore and that it is transferred at the end of the mating process. If mating can be interrupted early in the process, especially for EAB, which mates for an hour, fertilization will not be achieved. In addition, the researchers gained knowledge of dissection and staining techniques to examine the structure of spermatophore and sperm. We found that the structure of the sperm differed among species, with exotic EAB and another native, Agrilus obsoletoguttatus both having sperm bundled within the spermatophore and BBB and TLCB do not. The 2 species with bundled sperm had significantly longer mating times than species without bundled sperm. The study on relationship between the frequency of mating and fecundity in EAB and BBB will contribute the ability to rear EAB in captivity, an important component of biological control programs as well as research. This research will also add to the ability of modelers to predict the dynamics of the geographical spread of EAB. The information on BBB will be useful should BBB invade Europe. We continued biosurveillance of EAB throughout the state. This in turn may lead to early detection of EAB, an important piece of successful management of invasive species. In addition to potential detection of EAB, biosurveillance by C. fumipennis could detect other potentially invasive buprestids such as the European Oak Borer (Agrilus sulcicollis), which was recently detected in NY. The creation of a large network of volunteers, many active in garden clubs, arborist associations and the Master Gardener Program has enabled us to do more biosurveillance, and will spread awareness of EAB in particular and the threat of invasive species in general. The discovery that cuticular hydrocarbons are crucial components of the prey recognition system of C. fumipennis opens the way for further research into the identity of these hydrocarbons and eventual manipulation of the system. In particular, laboratory rearing of C. fumipennis could be facilitated by the ability to substitute prey items when buprestids are not available. The discovery of pathogens for Agrilus spp. could aid in the development of biological control programs for invasive Agrilus spp.
Publications
- Rutledge, C.E., Keena, M.A. 2012. Mating frequency and fecundity in the Emerald Ash Borer Agrilus planipennis (Coleoptera: Buprestidae). Ann. Entom. Soc. Am. 105(1):66-72
- Rutledge, C.E., Hellman, W., Teerling, C., Fierke, M.K. 2011. Two novel prey families for the buprestid-hunting wasp Cerceris fumipennis Say (Hymenoptera: Crabronidae). Coleopterists Bulletin 65(2):194-196
- Rutledge, C.E., Millar, J.G, Romero, C.M., Hanks, L.M. 2009. Identification of an important component of the contact sex pheromone of Callidiellum rufipenne (Coleoptera: Cerambycidae). Environ. Entomol. 38(4):1267-1275
- Rutledge, C.E., Keena, M.A. (2009) Mating Frequency and Fecundity in the Emerald Ash Borer. Emerald Ash Borer Research and Technology Development Meeting (abstracts). Forest Health Technology Enterprise Team. FHTET-2010-01 (P32)
- Hiskes R., Marra, R.E., Rutledge C.E. 2009. Invasive species and Connecticut's Forests. An introduction to their impact- potential and realized. Connecticut Woodlands 73(4):7-13 (non-refereed).
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Male Calidiellum rufipennis, the Small Japanese Cedar Borer (SJCB), release a volatile pheromone (3R)-3-hydroxy-2-hexanone. Both olfactometer and field-trapping tests using live males and a generic hydoxyketone mix showed that significantly more beetles landed on or were caught in traps with pheromone lures than in control traps. The traps attracted male and female beetles equally. We continued to examine the mating ecology of Agrilus anxius, the Bronze Birch Borer (BBB), A. planipennis, the Emerald Ash Borer (EAB), and A. bilineatus (Weber), the Twolined Chestnut Borer (TLCB). Sperm was compared among the species. In EAB, TLCB, and BBB, sperm is transferred in a spermatophore at the end of copulation. EAB bundle their sperm into large packets within the spermatophore, while BBB and TLCB do not. Sperm moves rapidly from the spermatophore into the spermatheca. The spermatophore is gone by 2-3 hours after mating. Sperm bundles of EAB do not unravel for 4-6 days; consistent with the observed time lag between mating and egg laying in EAB. EAB fecundity is increased by mating more than once, ensuring at least one successful mating, but no additional gain or loss is attached to mating more than twice. When allowed, males and females mated daily for a week. Mate choice was not influenced by a prior mating partner and individuals had no consistent preference for body size. Work on natural enemies of the Buprestidae concentrated on pathogens and on the wasp Cerceris fumipennis (Hymenoptera: Crabronidae). This wasp is a solitary predator that provisions her nest with adults of Buprestidae, including BBB, TLCB, and EAB when EAB is present. Surveys in 2010 located 18 additional colonies in CT (70 were identified in 2009). A network of 21 volunteers monitored colonies at 23 sites. The chemical basis of prey recognition of C. fumipennis was investigated. Stripping cuticular hydrocarbons renders buprestids unattractive to female wasps. Attraction was restored by reapplying the extracted hydrocarbons. Extracts from one genus restored attraction to another genus of Buprestidae, while extracts from a non-buprestid beetle did not. We found microsproridia in BBB. We observed a probable new species of microsporidia in Dicera lurida (Buprestidae) and work on the description and DNA sequencing of these organisms are ongoing. We also found pathogenic nematodes in these two species. The small subunit rDNA and Cytochrome (COX) regions of these nematodes were sequenced. Preliminary BLAST analysis of the ssrDNA shows the closest relationship (95-96% range) to the tylenchoid Deladenus siricidicola from the Siricid woodwasp, Sirex noctilio. PARTICIPANTS: Volatile pheromones of SJCB; Dr. Lawrence Hanks, UI Urbana-Champaign and Dr. Jocelyn Millar, UC Riverside. EAB mating and fecundity; Dr. Melody Keena, USFS in Hamden, CT. Biosurveillance was conducted with the help of 21 volunteer Wasp-Watchers. Host recognition of C. fumipennis; Dr. Peter Silk, Canadian Forest Service. Buprestid pathogens, Dr. Charles Vossbrinck, The Connecticut Agricultural Experiment Station. TARGET AUDIENCES: State and private foresters, nursery owners, green industry professionals, and home owners PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The role of the male-produced pheromone of C. fumipennis was defined as an aggregation pheromone. We ascertained that Agrilus anxius (BBB), A. planipennis (EAB), and A. bilineatus (TLCB) use a spermatophore that is transferred at the end of the mating process. If mating can be interrupted early in the process, especially for EAB, which mates for an hour, fertilization will not be achieved. In addition, the researchers gained knowledge of dissection and staining techniques to examine the structure of spermatophore and sperm. We found that the structure of the sperm differed between the exotic EAB and native species. The study of the relationship between frequency of mating and fecundity in EAB will contribute to the ability to rear EAB in captivity, an important component of biological control and research programs. This work also will add to the ability of modelers to predict the dynamics of the geographical spread of EAB. We continued biosurveillance of EAB throughout the state, which may lead to early detection of EAB, which is important in the successful management of invasive species. In addition to potential detection of EAB, biosurveillance by C. fumipennis could detect other potentially invasive buprestids such as the European Oak Borer (Agrilus sulcicollis), which was recently detected in NY. The creation of a large network of volunteers, many active in garden clubs, arborist associations, and the Master Gardener Program, has enabled us to increase biosurveillance activites with fewer resources and has increased public awareness of EAB and the threat of invasive species. The discovery that cuticular hydrocarbons are crucial components of the prey recognition system of C. fumipennis opens the way for further research into the identity of these hydrocarbons and eventual manipulation of the system. In particular, laboratory rearing of C. fumipennis could be facilitated by the ability to substitute prey items when buprestids are not available.
Publications
- No publications reported this period
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: A paper was published (see below) on the identification of an important component of the contact sex pheromone of Callidiellum rufipenne (Coleoptera: Cerambycidae) the Small Japanese Cedar Borer (SJCB). We investigated whether or not SJCB males use a trail pheromone to locate females as has been reported for other sub-families of Cerambycidae (e.g. Lamiinae: Anoplophora glabripennis). Several bioassays failed to show any evidence for this mate-location mechanism, which is consistent with behavior in other species in the sub-family of SJCB (Cerambycinae). The mating behavior of Agrilus anxius, the Bronze Birch Borer (BBB), A. planipennis the Emerald Ash Borer (EAB) and A. bilineatus (Weber), the Twolined Chestnut Borer (TLCB) were examined by detailed scoring of videotapes. The 3 species share a highly stereotyped mating behavior, and only differ in the duration of mating. EAB mates for an hour, while the two native species mate for only 5-10 minutes. In a subsequent experiment, in which mating pairs of EAB and BBB were disrupted at different points during mating, it was determined that spermatophore transfer to the female occurred at the end of the mating period in both species, despite the difference in mating duration. Spermatophore and sperm structure were compared, and it was found that EAB bundle their sperm into large packets within the spermatophore, while BBB do not. This information was disseminated via a talk at the annual meeting of the Eastern Branch of the Entomological Society of America and is in preparation for publication. We examined the impact of single versus multiple mating for female EAB and BBB longevity, fecundity and fertility. In the first two treatments, mated females were housed either alone (separate) or with their mate (together). In the third treatment, females were paired with a random male (unobserved). Lifetime fecundity and fertility were recorded. Results suggest a single mating may be sufficient to ensure maximal fitness for females, but there is a high potential for failure of any one mating, and no apparent cost to multiple mating. Thus, multiple mating is likely the best strategy for female EAB to maximize their fitness. Experiments with EAB are complete and in preparation for publication. BBB work is ongoing. This work was presented at the USDA EAB Research & Technology Development Meeting. Work on natural enemies of BBB, TLCB and EAB concentrated on the native predatory wasp Cerceris fumipennis (Hymenoptera: Crabronidae). The wasp is a solitary predator that provisions nests with adults of all species of the family Buprestidae. Surveys located 70 new colonies in Connecticut. BBB and TLCB are frequent prey of the wasp, and when EAB are present, the wasp readily preys on them. Work was done on the chemical basis of prey recognition of C. fumipennis for Buprestidae. Stripping cuticular hydrocarbons renders buprestids unattractive to female wasps. Attraction was restored by reapplying the extracted hydrocarbons. This was true with beetles in 4 different buprestid genera. Work will continue next summer to identify the cuticular compounds that wasps are using to identify buprestid prey. PARTICIPANTS: The work on the contact sex pheromone of SJCB was done in collaboration with Dr. Lawrence Hanks of the UI Urbana-Champaign and Dr. Jocelyn Millar of the UC Riverside. The work on EAB, and BBB mating and fecundity was done in collaboration with Dr. Melody Keena of USFS at Hamden CT. Work on the chemical basis of host recognition of Cerceris fumipennis was done in collaboration with Dr. Peter Silk of the Canadian Forest Service. TARGET AUDIENCES: The target audiences include state and private foresters, nursery owners, green industry professionals, and home owners. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The contact sex pheromone of C. rufipenne was identified. The synthesis of the pheromone, 5,13 dimethyl nonocosane was a novel synthesis. A description of the mating stereotyped mating behavior of 3 Agrilus spp. was gained. The discrepancy in mating duration between EAB and the native Agrilus spp. suggests that there are fundamental differences in their ecology. We also ascertained that this genus uses a spermatophore and that it is transferred at the end of the mating process. If mating can be interrupted early in the process, especially for EAB, which mates for an hour, fertilization will not be achieved. In addition, the researchers gained knowledge of dissection and staining techniques to examine the structure of spermatophore and sperm. The study on relationship between the frequency of mating and fecundity in EAB will contribute the ability to rear EAB in captivity, an important component of biological control programs as well as research. This research will also contribute to the body of literature addressing the role of female polyandry in the evolution of sex. Our location of 70 C. fumipennis colonies in Connecticut will allow for biosurveillance of EAB throughout the state. This in turn may lead to early detection of EAB, and important piece of successful management of invasive species. In addition to potential detection EAB, biosurveillance by C. fumipennis could detect other potentially invasive buprestids such as the European Oak Borer (Agrilus sulcicollis) which was recently detected in NY. The discovery that cuticular hydrocarbons are crucial components of the prey recognition system of C. fumipennis opens the way for further research into the identity of these hydrocarbons and eventual manipulation of the system. In particular, laboratory rearing of C. fumipennis could be facilitated by the ability to substitute prey items when buprestids are not available.
Publications
- Rutledge, C.E., Millar, J.G, Romero, C.M., Hanks, L.M. (2009)Identification of an important component of the contact sex pheromone of Callidiellum rufipenne (Coleoptera: Cerambycidae). Environ. Entomol. 38(4):1267-1275
- Hiskes R., Marra, R.E., Rutledge C.E. (2009) Invasive species and Connecticut's Forests. An introduction to their impact- potential and realized. Connecticut Woodlands 73(4):7-13 (non-refereed).
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: Objective 1. We investigated chemical aspects of host-finding strategies of Cerambycidae and Buprestidae. Bronze Birch Borer, Agrilus anxious Glory (Coleoptera: Buprestidae), BBB is a pest of birch trees in North America. BBB attacks stressed, living birches. The purple trap for Emerald Ash Borer (EAB) A. planipennis (Fairmaire), is baited with chemicals from stressed ash trees. We took bark samples of healthy and stressed grey birch (Betula populifolia) and collected their volatile emissions. These samples are currently being analyzed and compared using GC-MS. The Small Japanese Cedar Beetle, Callidiellum rufipenne (Motschulsky) (Coleoptera: Cerambycidae) (SJCB) is an invasive pest of dying trees in the cedar family. Adult beetles aggregate on the hosts of their larvae. SJCB males produce a volatile pheromone. In field trapping bioassays both sexes preferred traps baited with pheromone (82%) to controls (18%). In preliminary laboratory tests, females chose the scent of live males over fresh air. Information was disseminated to the general public via a poster at CT Ag. Exp. Station annual open house. Objective 2. We investigated mating behavior of Cerambycidae and Buprestidae. Male SJCB respond to females only on contact. Stripping cuticular hydrocarbons from females with solvent makes them unattractive. Reapplying solvent extract to washed females restores their attractiveness, confirming the role of cuticular hydrocarbons in mate recognition. Female cuticular hydrocarbon extracts contain dimethyl-5,7-nonacosane. We synthesized this compound and confirmed that it is the contact pheromone. Information has been shared verbally with foresters and via a poster at a CT. Ag. Exp. Station open house. Laboratory mating of BBB, EAB and A. bilineatus (Weber), the Twolined Chestnut Borer (TLCB) was recorded and scored in detail. All species shared a highly stereotyped mating behavior. EAB mated for 1 h and the two native species for under 10 m. Subsequently, mating pairs of BBB and EAB were separated at different time intervals and dissected. Both species transfer sperm in a spermatophore in the last stage of mating. Details of sperm structure of both species were examined. Information was disseminated via poster at the Entomological Society of America in 2007 and verbally at a workshop to 50 state foresters. Objective 3. We examined 10 logs to ascertain overwintering mortality in SJCB. Of 1023 holes examined we found 730 successful emergences (71%), and 293 (29%) dead adults in tunnels. Cause of death is unknown. We found 86 parasitoid cocoons, apparently all the same species. We are awaiting their identification. We dissected 50+ BBB adults from Connecticut paper birches (B. papayrifia). We found an unidentified microsporidian infesting a female. Its appearance is consistent with a newly discovered Cystosporogenes sp. infesting BBB in Canada (Kyei-Poku et al. 2008). We will confirm its identity using DNA. We found 2 BBB whose gonads were infested with unidentified nematodes. Information has been shared verbally with peers working in USDA EAB programs. PARTICIPANTS: Objective 1. Investigate chemical aspects of host-finding strategies of Cerambycidae and Buprestidae. Ms. Teri Arsenault of CT. Ag. Exp. Station is performing the GC-MS analysis of the birch samples comparing stressed and healthy trees. The work on SJCB pheromone was done in collaboration with Dr. Lawrence Hanks of the Department of Entomology of the University of Illinois Urbana-Champaign and Dr. Jocelyn Millar of the Department of Entomology of the University of California Riverside. Objective 2. Investigate mating behavior of Cerambycidae (C. rufipenne) and Buprestidae (A. anxius, A. bilineatus and A. planipennis) The work on the contact sex pheromone of SJCB was done in collaboration with Drs. Hanks and Millar (see above). The work on EAB mating was done in collaboration with Dr. Juli Gould of USDA PPQ-APHIS at Otis MA and Dr. Melody Keena of USFS at Hamden CT. Microscopy was aided by Dr. Theodore Andreadis of the CT Ag. Exp. Station. Observations of BBB were aided by Ms. Mioara Scott of the CT Ag. Exp. Station. Objective 3. Survey for natural enemies of Cerambycidae and Buprestidae The work on the microbial pathogens of BBB was done in collaboration with Dr. Andreadis (see above) and Dr. Charlie Vossbrinck of the CT. Ag. Exp. Station. TARGET AUDIENCES: The target audiences include state and private foresters, nursery owners, green industry professionals, and home owners. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Objective 1. We investigated chemical aspects of host-finding strategies of SJBC. Both male and female SJCB respond to a lure that attracts other Cerambycinae. Lures have been one of the most successful tools in wood boring insect management, and this discovery opens the way for development of new trapping technologies. Objective 2. The contact sex pheromone of C. rufipenne was identified. The synthesis of the pheromone, 5,13 dimethyl nonocosane was a novel synthesis. A description of the mating stereotyped mating behavior of 3 Agrilus spp. was gained. The discrepancy in mating duration between EAB and the native Agrilus spp. suggests that there are fundamental differences in their ecology. We also ascertained that this genus uses a spermatophore and that it is transferred at the end of the mating process. If mating can be interrupted early in the process, especially for EAB, which mates for an hour, fertilization will not be achieved. In addition, the researchers gained knowledge of dissection and staining techniques to examine the structure of spermatophore and sperm. Objective 3. In our survey for natural enemies, we determined that SJCB does not suffer great winter mortality, however we did confirm that this invasive beetle is being attacked by parasitoids. We found 2 microbial pathogens attacking BBB, one of which is a record for the United States, and the first recorded microsporidian in a species Buprestidae. Nematodes are also poorly known from Buprestidae, and there are few reports that are not related to the common entomopathogenic nematodes used in biological control. Investigation into these pathogens may be useful in EAB management.
Publications
- No publications reported this period
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