Attraction of the Larval Parasitoid Spintherus dubius (Hymenoptera: Pteromalidae) to Feces Volatiles from the Adult Apion Weevil Host.

The behavioral response of the larval parasitoid Spintherus dubius (Hymenoptera: Pteromalidae) to volatile compounds derived from its Apion weevil hosts was investigated in two-choice bioassays. Odor source candidates were the larval and adult stages of weevils, clover flowers, and feces from adult weevils. Despite S. dubius being a larval parasitoid, the odor of weevil larvae isolated from the clover flowers was not attractive to female parasitoids. Surprisingly, S. dubius females were instead attracted by the odor from the feces of adult weevils. The female parasitoids were similarly attracted to the feces produced by the two main hosts, the red clover weevil (A. trifolii) and the white clover weevil (A. fulvipes). Chemical analysis of the volatile composition of feces produced by the two hosts revealed qualitatively similar odor profiles, correlating with the observed attraction by the parasitoid towards both odor sources. Some of the identified volatile compounds are commonly present in clover plant headspace fractions and may function as a kairomone to facilitate orientation by S. dubius to Apion-infested clover flowers. Larval and adult weevils were not attractive for parasitoid females, whereas, for the white clover weevil-plant association, infested flowers were highly attractive. These data show the use by the clover weevil parasitoid of an alternative source of olfactory information for locating its host.

First Report of "Candidatus Liberibacter solanacearum" Associated with Psyllid-Affected Carrots in Sweden.

Carrot (Daucus carota) plants with symptoms resembling those associated with the carrot psyllid Trioza apicalis and the bacterium "Candidatus Liberibacter solanacearum" (1-4) were observed in 70% of commercial fields in southern Sweden in August 2011, with approximately 1 to 45% symptomatic plants per field. T. apicalis, a pest of carrot in northern and central Europe, including Sweden, can cause as much as 100% crop loss and is associated with "Ca. L. solanacearum" (1-4). Symptoms on affected plants include leaf curling, yellow and purple discoloration of leaves, stunted growth of shoots and roots, and proliferation of secondary roots (3). Carrot plant and psyllid samples were collected from fields in the province of Halland. Total DNA was extracted from petiole and root tissues of 33 symptomatic and 16 asymptomatic plants (cvs. Nevis and Florida), with the cetyltrimethylammonium bromide (CTAB) buffer extraction method (2,3). DNA was also extracted from 155 psyllids (3). DNA samples were tested by PCR using primer pairs OA2/OI2c (5''-GCGCTTATTTTTAATAGGAGCGGCA-3'/5'-GCCTCGCGACTTCGCAACCCAT-3') and CL514F/R (5'-CTCTAAGATTTCGGTTGGTT-3'/5'-TATATCTATCGTTGCACCAG-3'), to amplify a portion of 16S rDNA and rplJ/rplL ribosomal protein genes, respectively, of "Ca. L. solanacearum" (2,3). A 1,168-bp 16S rDNA fragment was detected in the DNA from all 33 symptomatic and two asymptomatic plants, and a 668-bp rplJ/rplL fragment was amplified from the DNA of all 33 symptomatic and four asymptomatic plants, indicating the presence of liberibacter. DNA from 23 and 49 psyllid samples yielded similar amplicons with OA2/OI2c and CL514F/R primer pairs, respectively. Amplicons from the DNA of four carrot roots and three T. apicalis with each primer pair were cloned (pCR2.1-TOPO; Invitrogen, Carlsbad, CA) and three clones of each of the 14 amplicons were sequenced (MCLAB, San Francisco, CA). BLAST analysis of the 16S rDNA consensus sequences from carrot (GenBank Accession No. JN863095) and T. apicalis (GenBank Accession No. NJ863096) showed 100% identity to those of "Ca. L. solanacearum" previously amplified from carrot (GU373048 and GU373049) and T. apicalis (GU477254 and GU477255) from Finland (2,3). The rplJ/rplL consensus sequences from carrot (GenBank Accession No. JN863093) and T. apicalis (GenBank Accession No. JN863094) were 99% identical to the sequences of rplJ/rplL "Ca. L. solanacearum" ribosomal protein gene from carrots in Finland (GU373050 and GU373051). To our knowledge, this is the first report of "Ca. L. solanacearum" associated with carrot and T. apicalis in Sweden. The disease associated with this bacterium caused millions of dollars in losses to potato and several other solanaceous crops in North and Central America and New Zealand (1). This plant pathogen is also associated with significant economic damage to carrot crops observed in Finland (2,3). References: (1) J. E. Munyaneza. Southwest. Entomol. 35:471, 2010. (2) J. E. Munyaneza et al. Plant Dis. 94:639, 2010. (3) J. E. Munyaneza et al. J. Econ. Entomol. 103:1060, 2010. (4) A. Nissinen et al. Entomol. Exp. Appl. 125:277, 2007.

Capture rates of the European pine sawfly, Neodiprion sertifer, in pheromone traps, with special regard to effects of wind speed.

Males of the European pine sawfly, Neodiprion sertifer Geoffr., were marked and released downwind from pheromone traps, baited with 100 micrograms of the sex pheromone (2S,3S,7S)-3,7-dimethyl-2-pentadecyl acetate. Males were released 5 m downwind from one trap, or downwind from five traps, 50 m or 200 m away. The average capture rates after 24 hr were 21.5%, 17.7% and 3.8%, respectively. The capture rate was highest at moderate wind speeds (1-2 m/sec) in the 50 m experiments, whereas it decreased above wind speeds of 1.5 m/sec in the 200 m experiments. With no precipitation and > 13.5 degrees C during overcast, wind speed is presumably the most important climatic factor for N. sertifer males flying upwind to a pheromone source. Travel time, the elapsed time form take-off to landing on the trap, varied considerably, and the shortest recorded travel times were 1, 6 and 45 min for the 5, 50, and 200 m experiments, respectively. The trap efficiency i.e., number of captured males per number of males that landed on the trap, was estimated at 52% in the 5 m experiments. The sampling range after 24 hr was calculated at approximately 400 m by regression analysis. The combination of the males' flight ability during upwind progress and their longevity (12 days), suggests a potentially large seasonal sampling range of the traps used in this study.

Release rates for pine sawfly pheromones from two types of dispensers and phenology of Neodiprion sertifer.

Comparisons of release rates, duration in the field, and catch efficiency of polyethylene and cotton roll dispensers for the sex pheromones of sawflies (Hymenoptera: Diprionidae) were conducted. The release rates of the Neodiprion sertifer (Geoffr.) and Diprion pini (L.) sex pheromones, the acetates of pentadecanol and (2S,3S,7S)-3,7-dimethyl (2S,3R,7R)-3,7-dimethyl-2-tridecanol from polyethylene dispensers were measured at different temperatures in the laboratory. The release rates for the substances depended on both the temperature and initial load in the vials. The catch from cotton rolls baited with 100 micrograms of the acetate or propionate of 3,7-dimethyl-2-pentadecanol was compared to the catch from regularly renewed cotton rolls baited with 10 micrograms of the same acetate. The catch was higher for the 100-microgram cotton rolls for, at most, 45 days, and there was no significant differences in catch between the acetate and the propionate. The catch in traps baited with polyethylene or cotton roll dispensers loaded with the acetate of 3,7-dimethyl-2-pentadecanol was compared and showed that cotton roll traps mirrored the decreasing release of the substance rather than the actual flight activity. The length of the flight period of N. sertifer in Sweden, the Czech Republic, Italy, and Greece did not exceed 100 days in any of the countries. By adjusting the initial pheromone load of the polyethylene vials to the expected temperatures, it should be possible to get a constant and sufficiently high release rate during the entire flight period.

Sex pheromone of the pine sawfly Macrodiprion nemoralis (Hymenoptera:Diprionidae): identification of (2S,3R,7R,9S)-3,7, 9-trimethyl-2-tridecanol as the precursor for the active pheromone acetate.

The main component of the sex pheromone precursor in females of Macrodiprion nemoralis was identified as a threo-3,7, 9-trimethyl-2-tridecanol isomer, approximately 800 pg per female, by gas chromatography-mass spectrometry. Comparison of mass spectrometric ion chromatograms showed that the natural compound in the female extract has the same retention time and mass spectrum as one of the two synthetic threo peaks. The acetate of the synthetic 16-isomer mixture caught a large number of males in the field, confirming the structure of the active pheromone. Comparison of gas chromatograms of the natural female extract with the eight synthetic threo stereoisomers showed that the pheromone is the (2S,3R,7R, 9S)-stereoisomer of 3,7,9-trimethyl-2-tridecyl acetate.

Field response of the pine sawflyNeodiprion sertifer to controlled release of diprionyl acetate, diprionyl propionate andtrans-perillenal.

Two dispenser types for pine sawfly (Hymenoptera: Diprionidae) pheromones-glass capillaries and dental cotton rolls-are described, and release rates reported. Glass capillaries of different diameters were used in field tests to determine dose-response relationships of (2S,3S,7S)-3,7-dimethylpentadecan-2-yl acetate (diprionyl acetate), diprionyl propionate, andtrans-perillenal on the trap catch of maleNeodiprion sertifer. A linear relationship between the logarithm of the capillary cross-sectional area and the logarithm of the release rate was found. However, capillaries of the same diameter could vary in the amount released by as much as a factor of 10. Both the acetate and propionate were attractive alone, but no synergistic effect was found.trans-Perillenal did not show any behavioral activity. Dental cotton rolls have been used extensively in diprionid pheromone research without knowledge about release rates. The release rate from rolls loaded with different amounts of diprionyl acetate was determined by extracting rolls after different times under constant temperature and wind conditions or after use in field tests. Quantification of diprionyl acetate was done on GC. The release rate was found to be proportional to the amount applied over several orders of magnitude, and persisted for several weeks. Capillaries and rolls with similar release rates caught similar numbers of sawflies.

Sex pheromone perception in male pine sawflies, Neodiprion sertifer (Hymenoptera; Diprionidae).

Electroantennographic and single sensillum recordings were performed on male pine sawfly, Neodiprion sertifer, antennae. Responses to the sex pheromone component (2S, 3S, 7S)- 3,7-dimethyl-2-pentadecenyl (diprionyl) acetate (SSS:OAc), to the behavioral inhibitor (2S, 3R, 7R)-diprionyl acetate (SRR:OAc), to the six other enantiomers of diprionyl acetate, and to the biosynthetic precursor diprionol were recorded. Responses to trans-perillenal, a monoterpene identified in female gland extracts and to (2S, 3S, 7S)-diprionyl propionate (SSS:OPr), a field attractant for N. sertifer and some related sawfly species were also recorded. EAG recordings demonstrated a high antennal sensitivity to SSS:OAc and to SSS:OPr. A somewhat lower response was elicited by SRR:OAc. Single sensillum recordings revealed 8-12 different cells firing in each sensillum, corresponding to the number of cells observed in earlier morphological investigations. Out of these cells all, except one, responded to SSS:OAc and to SSS:OPr. No differences in the response to the two components could be observed. The largest amplitude cell in each sensillum was specifically tuned to the behavioral antagonist, SRR:OAc. The pheromone perception system encountered in male pine sawflies thus differs clearly from that observed in moths.

Effective attraction radius : A method for comparing species attractants and determining densities of flying insects.

The catches of bark beetles (Coleoptera: Scolytidae) were compared between attractive traps releasing semiochemicals and passive traps (cylindrical sticky screens hung, at 10 heights of 0.7-11.5 m, on poles). A central attractive-trap pole was surrounded by three passive-trap poles spaced 50 or 100 m away at the apices of an equilateral triangle. The catches ofTomicus piniperda and other scolytid species on the attractive-trap pole baited with host monoterpenes, or the catches ofIps typographus attracted to synthetic pheromone, were compared to passive trap catches in a Scots pine forest or in a Norway spruce clear-cut, respectively. Information about flight height distributions of the above scolytid species, andHylurgops palliatus, Cryphalus abietis, Pityogenes chalcographus, P. quadridens, P. bidentatus, andTrypodendron domesticum were obtained on the passive and attractive trap poles. A new method is presented for determining the densities of flying insects based on the passive trap's dimensions and catch, duration of test, and speed of insect. Also, a novel concept, the effective attraction radius (EAR), is presented for comparing attractants of species, which is independent of insect density, locality, or duration of test. The EAR is obtained by the ratio of attractive and passive trap catches and the dimensions of the passive trap, and thus should correlate positively with the strength of the attractant and the distance of attraction. EARs are determined from catch data ofT. piniperda andI. typographus as well as from the data of previous investigations on the same or other bark beetles.

Attraction of bark beetles (Coleoptera: Scolytidae) to a pheromone trap : Experiment and mathematical models.

The movement of bark beetles near an attractive pheromone source is described in terms of mathematical models of the diffusion type. To test the models, two release experiments involving 47,000 marked spruce bark beetles [Ips typographus (L.)] were performed. The attractive source was a pheromone trap, surrounded by eight concentric rings with eight passive trap stations on each ring. Captures were recorded every 2-10 minutes for the pheromone trap and once for the passive traps. The models were fitted to the distribution in time of the central pheromone trap catch and to the spatial distribution of catch among the passive traps. The first model that gives a reasonable fit consists of two phases: Phase one-After release the beetles move according to a diffusion process with drift towards the pheromone trap. The strength of the drift is inversely proportional to the distance from the traps. Phase two-those beetles attracted to, but not caught by, the pheromone trap are no longer influenced by the pheromone, and their movement is described by a diffusion process without drift. In phase two we work with a loss of beetles, whereas the experiment seems to indicate that the loss of beetles in phase one is negligible. As a second model, the following modification of phase one is considered: After release the beetles move according to a diffusion process without drift, until they start responding to the pheromone (with constant probability per unit time), whereafter they start moving according to a diffusion process with drift. This study, like other release experiments, shows that the efficiency of the pheromone trap is rather low. What is specific for the present investigation is that we try to explain this low efficiency in terms of dynamic models for insect movement. Two factors seem to contribute: Some beetles do not respond to pheromone at all, and some beetles disappear again after having been close to the pheromone trap. It also seems that the motility of the beetles decreased after they ceased responding to the pheromone. Furthermore, the data lend some support to the hypothesis that flight exercise increases the response of the beetles to pheromone.