Aggregation pheromone of a newly described spruce bark beetle, Ips shangrila Cognato and Sun, from China

Volatiles from male hindgut extracts of a newly described spruce bark beetle, Ips shangrila Cognato and Sun, from different attack phases were analyzed by GC–MS/FID with both polar and enantioselective columns. The GC–MS/FID analyses showed that unmated males (Phase 1) or males mated with <3 females (Phases 2–4) produced 2-methyl-3-buten-2-ol and 99%-(+)-ipsdienol as major components, and (−)-cis-verbenol, (−)-trans-verbenol, myrtenol and 2-phenyl ethanol as minor or trace components. The release of these male-produced compounds was confirmed by the analysis on aeration sample of an I. shangrila infested wind-thrown spruce trunk. The quantities of 2-methyl-3-buten-2-ol, cis-verbenol and trans-verbenol from male hindgut extracts were almost unchanged or even slightly increased during gallery development, while ipsdienol decreased dramatically after mating with three females. No obvious Ips-related aggregation pheromone components were detected in the female hindgut extract. A field trapping bioassay in Qinghai, China, showed that the ternary blends containing 2-methyl-3-buten-2-ol, (−)-cis-verbenol and 97%-(+)-ipsdienol or (±)-ipsdienol, caught significantly more I. shangrila (♂:♀ = 1:2.14) than did the unbaited control. Replacing 97%-(+)-ipsdienol (close to the naturally produced enantiomeric ratio) with (±)-ipsdienol in the ternary blend seemed to reduce trap catches by 50%, but the difference was not statistically significant. Surprisingly, addition of (−)-trans-verbenol (at 0.2 mg/day) to the active ternary blends significantly reduced traps catches to the level not different from the unbaited control. Our results suggest that the two major components, 2-methyl-3-buten-2-ol and 99%-(+)-ipsdienol, plus a minor component, (−)-cis-verbenol, produced by fed males, are likely the aggregation pheromone components of I. shangrila.     

The aggregation pheromone ofIps duplicatus and its role in competitive interactions withI. typographus (Coleoptera: Scolytidae)

Summary Ips duplicatus withI. typographus co-inhabiting Norway spruce (Picea abies (L.) Karst.) would benefit from a pheromone blend distinct from that of the larger competitorI. typographus. GC-MS analysis showed thatI. duplicatus males feeding in the host produced ipsdienol (Id),cis-verbenol (cV),trans-verbenol (tV), myrtenol (Mt), andE-myrcenol (EM) and traces of 2-methyl-3-buten-2-ol (MB).I. duplicatus produced Id in approximately racemic form (48.9-54.5% (+)-(S)-isomer). The amounts of Id and EM released over a 9 day period had a maximum of 250 and 5 ng/h/male, respectively, on day 2. Exposure ofI. duplicatus males to myrcene and -pinene resulted in the production of small amounts of Id, cV, tV, Mt, andtrans-pinocarveol, but not of EM. In laboratory bioassays with walking beetles, the pheromone component Id alone was weakly attractive while EM was inactive, but in binary combination with Id strongly synergized attraction. A combination of EM and Id at a release rate equivalent to 100–200 males was more attractive in the field than 70 unmated males in a spruce log. The addition of myrcene ( a suggested pheromone precursor of Id) to Id did not enhance trap caches, while addition of EM increased catches > 10-fold. Subtracting EM from a blend of Id, EM, cV and MB drastically reduced trap catches while subtraction of cV or MB or both had no significant effect. Addition of EM over a wide concentration range to the synthetic pheromone ofI. typographus did not reduce the attraction of females of this species in the laboratory. A two-species pheromone interaction field test releasingI. typographus pheromone components (MB + cV) at 10–1000 male equivalents (ME) andI. duplicatus pheromone (Id + EM) at 0, 10–1000 ME in all possible combinations showed both positive intraspecific dose-response effects and an interspecific inhibition. Higher release rates of EM appeared to inhibitI. typographus, especially males. In a tree colonization model, the response of the two competing species to their respective pheromones show a good separation during the mass-attack with a small initial cross-attraction. It remains to be shown whether either of the two pheromone systems have in fact evolved in the present sympatry, or if they are an incidental effect of ancestry of these phylogenetically distantIps.     

Identification of the putative aggregation pheromone components emitted by the European earwig, <Emphasis Type="Italic">Forficula auricularia</Emphasis>

The European earwig, Forficula auricularia is an invasive insect found in many temperate regions of the world and is regarded as an urban and agricultural pest of numerous crops. Several studies have shown that F. auricularia aggregate in large numbers with the use of an aggregation pheromone. However, the identity of compounds which constitute the pheromone remains elusive. Our aim was to isolate and identify the aggregation pheromone used by F. auricularia using both solvent washes of individuals and thermal desorption of substrates exposed to earwigs. Solvent washes of male, female and juvenile earwigs isolated 51 different branched and unbranched alkanes, alkenes and alkadienes. Substrates exposed to aggregating field populations in situ were demonstrated to be attractive to earwigs after less than 24 h of exposure. Analysis of these substrates using thermal desorption and solvent washes showed that hydrocarbons were the only detectable compounds laid down by earwigs on these surfaces. Significant behavioural responses were observed to synthetic blends of the unsaturated hydrocarbons containing (Z)-7-tricosene, (Z)-9-tricosene, (Z)-7-pentacosene and (Z)-9-pentacosene at ≥25 insect equivalents in field-based bioassays. However, behavioural responses to these blends proved inconsistent particularly later in the field season, possibly due to a missing component within the pheromone blend or plasticity in the pheromones production and response.     

GC-EAD responses to semiochemicals by eight beetles in the subcortical community associated with Monterey pine trees in coastal California: similarities and disparities across three trophic levels

Summary.  Antennae of six sympatric bark and ambrosia beetles (Scolytidae), Dendroctonus valens LeConte, Gnathotrichus retusus (LeConte), Hylastes tenuis Eichhoff, Ips mexicanus (Hopkins), Ips plastographus maritimus Lanier, and Pseudohylesinus sericeus (Mannerheim), and two scolytid predators, Enoclerus sphegeus (F.) (Cleridae) and Lascontonus tuberculatus Kraus (Colydiidae), were analyzed by gas chromatographic-electroantennographic detection (GC-EAD) for their responses to synthetic Ips spp. pheromone components, and host and nonhost volatiles. The beetles emerged from cut logs of pitch canker-infected Monterey pine trees, Pinus radiata D. Don. There were significant disparities in EAD response patterns to the hemiterpene and monoterpene alcohol pheromone components that are typically produced by Ips spp. Antennae of I. p. maritimus responded strongly to ( ± )-ipsdienol, ( ± )-ipsenol, amitinol, and lanierone; antennae of I. mexicanus responded strongly to (1S,2S)-(–)-cis-verbenol, with weaker responses to ( ± )-ipsdienol, ( ± )-ipsenol, and amitinol; antennae of H. tenuis responded to (1S, 2R)-(–)-trans-ver-benol, with less pronounced responses to (–)-cis-verbenol and 2-methyl-3-buten-2-ol; and antennae of D. valens, G. retusus, and P. sericeus generally responded to all Ips spp. pheromone components except 2-methyl-3-buten-2-ol (D. valens and G. retusus) and E-myrcenol (G. retusus and P. sericeus). Ips mexicanus responded only to the (–)-enantiomers of ipsenol and ipsdienol, whereas I. p. maritimus responded to (–)-ipsenol, but to both the (+)- and (–)-enantiomers of ipsdienol. The antennae of the two predaceous insects (E. sphegeus and L. tuberculatus) responded to a range of the Ips spp. pheromone components. Host monoterpenes elicited no antennal responses from E. sphegeus, G. retusus, H. tenuis, and I. mexicanus, but several monoterpenes elicited various levels of responses from D. valens and I. p. maritimus antennae. Interestingly, antennae of female D. valens responded to (–), but not (+)-limonene. α- and β-Pinene elicited weak responses from L. tuberculatus antennae. EAD responses to selected nonhost volatiles were almost identical among the six scolytid species, with trans-conophthorin eliciting the strongest response in most cases, followed by three C₆- alcohols and two C₈-alcohols. The antennal responses by most of these species to linalool or geranylacetone were very weak; (E)-2-hexenal, (Z)-3-hexenyl acetate, and benzyl alcohol elicited almost no response. The response pattern of P. sericeus to nonhost volatiles differed slightly from the rest of the scolytids: a strong response to linalool, weaker response to the C₈-alcohols. The two predaceous Coleoptera generally had weak, but detectable, responses to nonhost volatiles, except for a relatively strong response to trans-conophthorin by L. tuberculatus. No notable differences in EAD responses were observed between males and females of the two Ips spp. Our results provide an electrophysiological baseline for future efforts to identify attractive and repellent semiochemicals (aggregation pheromones, host kairomones, or nonhost interruptants) for this guild of scolytids and their key predators that are associated with moribund and pitch canker- infected P. radiata.     

Heritable pheromone blend variation in a local population of the butterbur borer moth Ostrinia zaguliaevi (Lepidoptera: Crambidae)

Summary. In a local population of Ostrinia zaguliaevi Mutuura & Munroe (Lepidoptera: Crambidae), extensive variation was found in the blend ratio of three sex pheromone components, (Z)-9-tetradecenyl acetate (Z9-14:OAc, 10.2-63.8%), (Z)-11-tetradecenyl acetate (Z11-14:OAc, 32.2-86.8%), and (E)-11-tetradecenyl acetate (E11-14:OAc, 2.1-11.9%). The variation was observed over a three-year period (2002-2004). Mother-daughter regression analyses have shown that although the heritability of the minor component E11-14:OAc was not significant, the heritability of the proportions of Z9-14:OAc and Z11-14:OAc were substantial (0.5–0.6). In artificial selection experiments, the mean % Z9-14:OAc in the sex pheromone changed significantly within three generations (37% in the control line, 48 and 52% for two lines selected for increase, and 23 and 30% for two lines selected for decrease). Despite these changes, the amounts of fatty acyl pheromone precursors, (Z)-9-, (Z)-11- and (E)-11-tetradecenoate, in the pheromone gland were not significantly affected by the selection. Taken together, variation in the pheromone blend of O. zaguliaevi is likely to be attributable to a few genes involved in the reduction or acetylation of fatty acyl pheromone precursors, the last two steps in pheromone biosynthesis.     

Phenylacetonitrile is a male specific repellent in gregarious desert locusts, Schistocerca gregaria

Summary. The behavioural response of desert locust individuals to adult male volatiles had been investigated in a dynamic Y-T-olfactometer. Synthetic blends of a postulated adult male-produced aggregation pheromone with or without phenylacetonitrile were not attractive to fifth-instar nymphs, young or mature adults of both sexes. Instead phenylacetonitrile had a repellent effect, which was strongest in sexually mature males. The repellent effect of phenylacetonitrile was not modified by the other components of the adult male pheromone bouquet. Even at low concentrations tested the source of the pheromone was avoided. These results are in agreement with recent studies showing that this male specific pheromone is employed by mature gregarious males in sexual behaviour and acts as a courtship inhibiting pheromone. Low concentrations of phenylacetonitrile or the postulated adult male aggregation pheromone blend without phenylacetonitrile may signal the presence of swarm fellows and eventually initiate visually guided congregation behaviour in gregarious locusts. Whether this effect does significantly contribute to swarm cohesion needs further studies.     

Population differentiation in female sex pheromone and male preferences in a solitary bee

Population differentiation in female mating signals and associated male preferences can drive reproductive isolation among segregated populations. We tested this assumption by investigating intraspecific variation in female sex pheromone and associated male odour preferences among distant populations in the solitary bee Colletes cunicularius (L.) by using quantitative gas chromatography and by performing field bioassays with synthetic blends of key sex pheromone compounds. We found significant differences in sex pheromone blends among the bee populations, and the divergence in odour blends correlated positively with geographic distance, suggesting that genetic divergence among distant populations can affect sex pheromone chemistry. Our behavioural experiments, however, demonstrate that synthetic copies of allopatric female sex pheromones were cross-attractive to patrolling males from distant populations, making reproductive isolation by non-recognition of mating signals among populations unlikely. Our data also show that patrolling male bees from different populations preferred odour types from allopatric populations at the two sites of bioassays. These male preferences are not expected to select for changes in the female sex pheromone, but may influence the evolution of floral odour in sexually deceptive orchids of the genus Ophrys that are pollinated by C. cunicularius males.     

Aggressive chemical mimicry of moth pheromones by a bolas spider: how does this specialist predator attract more than one species of prey?

Summary. The bolas spider, Mastophora hutchinsoni, attracts Lacinipolia renigera and Tetanolita mynesalis males by mimicking the female moth sex pheromones. However, as the prey species use completely different pheromone blends we conducted experiments to determine how this is accomplished by the predator. The periodicity of L. renigera mate-seeking activities occurs early in the scotophase, whereas male T. mynesalis are active late at night, corresponding with periods when these moths are captured by the spider. The pheromone blend of early-flying L. renigera interferes with attraction of late-flying T. mynesalis to its pheromone in a dose-dependent manner, suggesting the spider must always produce a single sub-optimal “compromise” blend for both species or that it adjusts its allomonal blend to optimize capture of the respective prey species at different times during the night. We delayed (L. renigera) or advanced (T. mynesalis) the periodicity of male activity through photoperiodic manipulation and found that the bolas spider attracted both prey species outside their normal activity windows. These results support the idea that bolas spiders produce components of both species at all times rather than producing the pheromone of each prey species at different times of the night. However, using coupled gas chromatography-electroantennography, we also demonstrated that the spider decreases its emission of the L. renigera pheromone over the course of the night. This modification should reduce the behavioral antagonism of the L. renigera pheromone on T. mynesalis males and increase the predator's success of attracting T. mynesalis during this prey's normal activity window late at night. Received 13 October 2001; accepted 28 December 2001.     

Host-plant green-leaf volatiles synergize the synthetic sex pheromones of the corn earworm and codling moth (Lepidoptera)

Summary The capture of adult male moths in female sex pheromone traps of two key agricultural pests, the corn earworm (Helicoverpa zea) and the codling moth (Cydia pomonella), is enhanced or synergized by a certain group of host-plant volatiles, the green-leaf volatiles (GLVs). Since female adults of both species call and release their sex pheromones while perched upon the leaves of their host-plants, the volatile constituents from the leaves of a number of host-plants were compared. Sex pheromone traps containing one of the prominent leaf volatiles of certainH. zea hosts, (Z)-3-hexenyl acetate, not only significantly increased the capture ofH. zea males but were preferred over traps baited only with sex pheromone. Similarly, traps baited with synthetic sex pheromome ofC. pomonella plus a blend of GLVs captured significantly more males than traps baited only with sex pheromone. Since male moths are not captured in traps baited only with these GLVs, it appears that these GLVs act as pheromone synergists which increase or enhance the attraction or arrestment of male moths in pheromone traps.     

Predicting the Probability of Outbreeding Depression

Abstract: Fragmentation of animal and plant populations typically leads to genetic erosion and increased probability of extirpation. Although these effects can usually be reversed by re‐establishing gene flow between population fragments, managers sometimes fail to do so due to fears of outbreeding depression (OD). Rapid development of OD is due primarily to adaptive differentiation from selection or fixation of chromosomal variants. Fixed chromosomal variants can be detected empirically. We used an extended form of the breeders’ equation to predict the probability of OD due to adaptive differentiation between recently isolated population fragments as a function of intensity of selection, genetic diversity, effective population sizes, and generations of isolation. Empirical data indicated that populations in similar environments had not developed OD even after thousands of generations of isolation. To predict the probability of OD, we developed a decision tree that was based on the four variables from the breeders’ equation, taxonomic status, and gene flow within the last 500 years. The predicted probability of OD in crosses between two populations is elevated when the populations have at least one of the following characteristics: are distinct species, have fixed chromosomal differences, exchanged no genes in the last 500 years, or inhabit different environments. Conversely, the predicted probability of OD in crosses between two populations of the same species is low for populations with the same karyotype, isolated for <500 years, and that occupy similar environments. In the former case, we recommend crossing be avoided or tried on a limited, experimental basis. In the latter case, crossing can be carried out with low probability of OD. We used crosses with known results to test the decision tree and found that it correctly identified cases where OD occurred. Current concerns about OD in recently fragmented populations are almost certainly excessive.     

Dynamics of pheromone production and communication in the mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini (Say) (Coleoptera: Scolytidae)

Summary. The mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini (Say), often co-exist in lodgepole pine, Pinus contorta var. latifolia Engelmann. Intra- and interspecific semiochemical communication occurs in both species and their complete semiochemical repertoire and precise dynamics of pheromone production have not been elucidated. Porapak-Q extracts of captured volatiles from beetles of each species aerated at different attack phases (freshly emerged, pioneer sex alone in the log and both sexes paired in new galleries), followed by gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectroscopic analyses identified 17 compounds (seven compounds common to both species, six present in D. ponderosae and four present in I. pini) that excited the antennae of either or both species. Seven compounds for D. ponderosae and nine for I. pini had not been assessed for behavioural activity. In field trapping experiments, 2-phenylethanol produced by both species inhibited the response of D. ponderosae to its aggregation pheromones. exo- and endo-Brevicomin produced by D. ponderosae significantly decreased the response of I. pini to its aggregation pheromone ipsdienol. Nonanal, a ubiquitous compound found in the volatiles of lodgepole pine, various nonhosts and in both beetle species deterred the response of I. pini to ipsdienol. The occurrence of cis-verbenol, trans-verbenol and verbenone in emergent I. pini, and verbenone and 2-phenylethanol in emergent D. ponderosae suggests that these compounds may inhibit aggregation and induce dispersal following emergence. Termination of aggregation in D. ponderosae appears to depend on the production of frontalin in combination with changes in the relative ratios of verbenone, exo-brevicomin, trans-verbenol and 2-phenylethanol. In I. pini, the cessation of ipsdienol production by males is probably the main factor in terminating aggregation. Received 16 November 1999; accepted 7 August 2000     

Experience counts: females favor multiply mated males over chemically endowed virgins in a moth (<Emphasis Type="Italic">Utetheisa ornatrix</Emphasis>)

Determining the factors that affect male mating success is essential to understanding how sexual selection operates, including explanations of the adaptive value of female preferences and how variation in male traits is maintained in a population. Although females may appear to choose males based on a single parameter, female mate choice is often a complex series of assessments of male quality that can only be revealed through manipulation of multiple male traits. In the moth Utetheisa ornatrix (Lepidoptera: Arctiidae), females have been shown to judge males primarily on their production of a courtship pheromone, hydroxydanaidal, derived from defensive chemicals acquired as larvae. Recent work, however, suggested that other factors, including prior mating experience by males, may also influence the outcome of precopulatory interactions with females. I ran mating trials with one female and two males to determine whether there were any differences in male mating success based on their prior exposure to females, mating experience, and time between matings. Previously mated males were favored over virgins when both males lacked the pheromone, but courting experience and mating interval did not explain these differences in male mating success. Furthermore, multiply mated males lacking the pheromone were favored over virgin males that produced the pheromone, thus reversing the commonly observed trend of female precopulatory bias towards males with higher levels of the pheromone. These results demonstrate that males with mating experience can secure copulations despite deficiencies in the pheromone, and I provide possible mechanisms and discuss their implications regarding sexual selection.     

Stereoisomeric composition of the sex pheromone of the pine sawfly <Emphasis Type="Italic">Microdiprion pallipes</Emphasis> (Hym., Diprionidae)

Summary. The basic chemical structure of the sex pheromone of the pine sawfly Microdiprion pallipes (Fallén) has earlier been identified as the propionate ester of (2S,3S,7R/S,11R/S)/(2R,3R,7R/S,11R/S)-3,7,11- trimethyl-2-tridecanol. We now report the results from further investigations on the male response to individual stereoisomers and to blends of stereoisomers, both in electroantennographic (EAG) recordings and in field trapping experiments. We also present our attempts to determine the stereochemistry of the compounds present in females of M. pallipes. By comparing gas chromatograms and mass spectra obtained from natural extracts with those from synthetic compounds it was found that the females contain one or more of the four (2S,3S,7R/S,11R/S)-3,7,11-trimethyl-2- tridecanol isomers (SS++-1). The active pheromone component is the corresponding propionate ester 2. In EAG experiments, males responded most strongly to five propionate ester samples, namely two four-isomer blends: SS++-2 and SR++-2, and three individual stereoisomers: SSSR-, SRRR- and SRSR-2. In a series of field trapping experiments it was found that males were attracted to the SR++-2 four-isomer blend and to the individual isomer SSSR-2. Based on the EAG-recordings and field responses of males and the stereoisomers found in the females, we suggest that the propionate ester of (2S,3S,7S,11R)-3,7,11-trimethyl-2- tridecanol (SSSR-2) is used as a main component of the sex pheromone in M. pallipes. Apparently the males react to other stereoisomers in addition to that or those produced by the females.     

Differential activity of non-fluorinated and fluorinated analogues of the European corn borer pheromone

Summary.  The differing antagonist activity of (Z)-13-hexadecen-2-one (Z11 – 14 :MK, 1) and its 1,1,1-trifluoro derivative (Z11 –14 :TFMK, 2), two closely related analogues of the European corn borer pheromone Ostrinia nubilalis (Z strain), and their rationale is reported. Both chemicals exhibited some electrophysiological activity, and topical application of 10 pg of pheromone analogue on male antennae was sufficient to induce significantly lower depolarization responses to the pheromone versus untreated insects. In a wind tunnel, the number of European corn borer males attracted to sources containing mixtures of 1 + pheromone in ratios ≥ 1 :1 was significantly lower than the number attracted to a source containing pheromone alone. Source contact behaviour was dramatically impaired when the 1 + pheromone blend reached a ratio of 10 :1, in which only 2% of males displayed source contact in the presence of antagonist. When compound 1 was present at the source, males usually flew upwind with occasional downwind reversals; when compound 2 was present at the lure, males performed wider crosswind reversals, with little progress toward the source. In the field, traps baited with mixtures of both compounds with the pheromone in ratios of 5 :1 and 10 :1 elicited a significantly decreased number of male catches. In esterase inhibition assays, compound 2 was a potent inhibitor (IC₅₀ = 70 nM), whereas the non-fluorinated compound 1 was not. The different activity of both compounds is presumed to be due to different mechanisms of action; considerations for using methyl ketone analogues as new behavioural antagonists of the pheromone are outlined.     

Reversible royalty in worker honeybees (<Emphasis Type="Italic">Apis mellifera</Emphasis>) under the queen influence

In most social insects, worker sterility is reversible, and in the absence of the queen, at least some workers develop ovaries and lay male-destined eggs. In the honeybee, reproductive workers also produce queen-characteristic mandibular and Dufour’s pheromones. The evolution of worker sterility is still under debate as to whether it is caused by queen manipulation (queen-control hypothesis) or represents worker fitness maximization (worker-control hypothesis). In this study, we investigated whether worker fertility and royal pheromone production are reversible under the queen influence. To that effect, we induced ovary activation and queen pheromone production in workers by rearing them as queenless (QL) groups. These workers were subsequently reintroduced into queenright (QR) microcolonies for 1 week, and their ovary status and queen pheromone levels were monitored. Workers reintroduced into QR, but not QL colonies, showed a clear regression in ovary development and levels of the queen pheromones. This is the first demonstration that worker sterility and/or fertility is reversible and is influenced by the queen. These results also emphasize the robustness of the coupling between ovary activation and royal pheromone production, as well as lending credence to the queen-control hypothesis. The dynamics of queen pheromone production in QL workers supports the role of Dufour’s gland pheromone as a fertility signal and that of the mandibular gland pheromone in dominance hierarchies.The two authors, Osnat Malka and Shiri Shnieor, contributed equally to this work.     

Comparative studies on the sex pheromones of Ostrinia spp. in Japan: the burdock borer, Ostrinia zealis

Summary. To gain insight into the evolution of the sex pheromone communication system in Ostrinia (Lepidoptera Pyralidae), the sex pheromone of the burdock borer, O. zealis was analyzed by means of gas chromatography-electroantennographic detection (GC-EAD), GC-mass spectrometry and a series of bioassays. Four EAD-active compounds were detected in the female sex pheromone gland extract, and these were identified as tetradecyl acetate (14:OAc), (Z)-9-tetradecenyl acetate (Z9–14:OAc), (E)-11-tetradecenyl acetate (E11-14:OAc) and (Z)-11-tetradecenyl acetate (Z11-14:OAc). The average amounts (ratio) of the four compounds in single sex pheromone glands were 2.5 ng (13%), 11.6 ng (61%), 4.1 ng (21%) and 0.9 ng (5%), respectively. In a wind-tunnel bioassay, the ternary blend of Z9-, E11- and Z11-14:OAc at a ratio found in the sex pheromone gland elicited the same behavioral responses from the males as did virgin females. 14:OAc did not show any enhancement or inhibition of the males’ behavioral responses when added to the ternary blend. The attractiveness of the 3-component lure to O. zealis males was also confirmed by field trapping experiments. Based on these results, we concluded that the sex pheromone of O. zealis is composed of Z9-14:OAc, E11-14:OAc and Z11-14:OAc at a ratio of 70:24:6. The evolutionary changes of the sex pheromones in Ostrinia are also discussed based on the presently available information on the sex pheromones and phylogenetic relationships of Ostrinia spp. Received 25 September 1998; accepted 2 December 1998.     

Inbreeding and Outbreeding Depression in Natural Populations of Chamaecrista fasciculata (Fabaceae)

Abstract: The deleterious consequences of inbreeding have been well documented. There are, however, few empirical studies that have examined the consequences of restoring heterozygosity and hence the fitness of inbred populations by conducting interpopulation crosses and measuring the performance of later-generation hybrids under field conditions. We conducted interpopulation crosses of 100 m to 2000 km, which spans the range of Chamaecrista fasciculata ( Fabaceae) in eastern North America. We then contrasted the performance of the F1 and later-segregating F3 hybrids with the parental generation. We found almost universal F1 superiority over the parents. The F3 hybrids suffered a loss of fitness compared to the F1 hybrids. The drop off in fitness of the F3 reflects both the loss of heterozygosity and the disruption of coadapted gene complexes. The F3 performance, however, was still often equal to that of the parents, suggesting that heterosis can outweigh the loss of coadaptation except for the longest-distance crosses. In a subset of environments, the F3 performance of long-distance (≥1000 km) interpopulation crosses was less than that of both parents and indicated true outbreeding depression. For C. fasciculata , it appears that crossing populations of up to intermediate distances of hundreds of kilometers has a short-term beneficial effect on progeny performance through F1, and that longer-term effects are not necessarily disruptive of fitness, at least relative to parental performance. The degree of F1 heterosis and F3 outbreeding depression varied between site and year, however, indicating an important role for the environment in the expression of these effects.     

Doubly uniparental transmission of mitochondrial DNA length variants in the mussel <Emphasis Type="Italic">Mytilus</Emphasis><Emphasis Type="Italic">trossulus</Emphasis>

Doubly uniparental inheritance of mitochondrial DNA (mtDNA) is an unusual feature found in marine mussels and a few other bivalve species. In addition to a mitochondrial genome (F) inherited through the female line, heteroplasmic individuals (males) contain a second highly diverged genome (M) that is inherited through the paternal line. The Baltic mussel Mytilus trossulus is characterized by the presence of two phylogenetically close female and male genomes. We examined M. trossulus sampled from the Gulf of Gdansk, southern Baltic. Somatic tissues and gametes were surveyed for the presence of different types of mtDNA. Length variants were identified using PCR (polymerase chain reaction) amplification of the mtDNA fragment containing the major noncoding region. Seventeen length variants present in homo- and heteroplasmic individuals were found in a sample of 343 individuals analyzed. Two length variants of the major noncoding region (PCR product lengths: 1520 and 1370 bp) were the most frequent. Length heteroplasmy was found in 46.8% of males, and was mostly caused by the presence of the short length variant (PCR product length: 1370 bp) of the major noncoding region. MtDNA variants were also detected by restriction analysis of a 1.3 kb segment of coding region (ND2-COIII) amplified by PCR. This study provides evidence that the short length variant is always transmitted to sperm and has taken over the role of the M genome. The remaining length variants were present in both males and females. Longer length variants were transmitted mainly through the female line. A possibility of much higher incidence of genome role reversals in Baltic M. trossulus, in comparison with other populations of mussels is discussed.     

Reevaluation of the sex pheromone of the lesser date moth, Batrachedra amydraula, using autosampling SPME-GC/MS and field bioassays

In a previous study of the sex pheromone of the lesser date moth, Batrachedra amydraula, using laboratory females, (Z4,Z7)-4,7-decadien-1-yl acetate, Z4-decen-1-yl acetate, Z5-decen-1-yl acetate and decyl acetate were identified by sequential SPME-GC/MS analysis. Traces of Z5-decen-1-ol were detected only in airborne collections. Concomitant field tests and re-evaluation of the composition of the sex pheromone of B. amydraula with feral female moths, using an improved sequential SPME-GC/MS procedure revealed a complex mixture of candidate pheromonal compounds. The same unsaturated acetates and the corresponding alcohols: (Z4,Z7)-4,7-decadien-1-ol, Z4-decen-1-ol, Z5-decen-1-ol were positively identified. In addition, the corresponding aldehydes, octanol and octyl acetate were also detected. All compounds were found to be released in a circadian rhythm, in a narrow time window of 2 h, approximately 1 h before sunrise. Comprehensive field bioassays indicated that the optimal attractive blend is a three-component mixture of (Z4,Z7)-4,7-decadien-1-yl acetate, Z5-decen-1-yl acetate and Z5-decen-1-ol in a ratio of 1:2:2. This blend gave about fivefold higher trap catch of B. amydraula males as compared to the previously published binary blend of (Z4,Z7)-4,7-decadien-1-yl acetate and Z5-decen-1-yl acetate in a ratio of 1:2. The alcohol Z5-decen-1-ol is an essential synergistic component of the sex pheromone of B. amydraula. All other identified compounds are inert, being neither synergists nor inhibitors of the pheromone. The optimal pheromone blend will be a useful tool in monitoring and control of B. amydraula, which is a serious pest of date plantations throughout the Middle East and northern Africa.     

Identification of the sex pheromone of Conogethes pluto: a pest of Alpinia

In recent years, Conogethes pluto (Lepidoptera: Crambidae) has become a major pest of Alpinia and other ornamental gingers in the Northern Territory and Queensland, Australia. This pest damages the flowers and bores into the stems, causing substantial losses to production. Currently, no synthetic sex pheromone is available to monitor or control this pest. This work aims at the identification of the sex pheromone of this pest. Analysis of the sex pheromone gland of female C. pluto by gas chromatography/electroantennogram detector revealed the presence of seven candidate pheromone compounds that elicited electroantennogram responses. Using gas chromatography/mass spectrometry analysis and micro-derivatization reactions, six compounds were identified as (E)-10-hexadecenal, as the main pheromone compound, (Z)-10-hexadecenal, hexadecanal, (E)-10-hexadecen-1-ol, (10E,12E)-hexadeca-10,12-dienal and (3Z,6Z,9Z)-tricosa-3,6,9-triene as minor pheromone compounds. In two-field trapping experiments, C. pluto responded to the six-component blend, and three of six compounds, i.e., (E)-10-hexadecenal, (3Z,6Z,9Z)-tricosa-3,6,9-triene, and (10E,12E)-hexadeca-10,12-dienal were shown to be necessary for attraction. In a subsequent experiment testing various doses (i.e., 0.01, 0.1, and 1 mg) of the six-component blend, the largest number of males was captured in traps baited with a lure loading of 1 mg. The availability of the sex pheromone of C. pluto will enable monitoring and provides the basis for additional control options for this pest.