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1.
A diagnostic polymerase chain reaction (PCR) method is presented for differentiating among the North American internal apple-feeding pests codling moth, Cydia pomonella (L.); oriental fruit moth, Grapholita molesta (Busck); lesser appleworm, Grapholita prunivora (Walsh); and cherry fruitworm, Grapholita packardi Zeller. An approximately 470-bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in three to six specimens of each species. Consistent and diagnostic differences were observed among the species in two regions of COI from which forward and reverse primers were designed to amplify a 112-116-bp segment of the gene. The primer sets were used to selectively amplify DNA from specimens of diverse geographic origin for each corresponding target species. Protocols were adapted for conventional and quantitative PCR, the latter being substantially faster. The method was validated as a decision-making tool for quarantine identifications for Mexico by representatives of their phytosanitary agency (Sanidad Vegetal). The method can facilitate identification of intercepted internal feeding Lepidoptera in apple and pear for many other importing nations.  相似文献   

2.
This study examined cross-species amplification of 33 microsatellite markers, previously developed for Cydia pomonella, in three related fruit moth species of the same tribe (Grapholitini), namely Grapholita molesta, Grapholita funebrana and Grapholita lobarzewskii. Eight microsatellite loci yielded polymorphic products for Gmolesta, nine for Gfunebrana and 11 for Globarzewskii. At all these loci, the number of alleles ranged between four and 11 in Gmolesta, and between four and nine in Gfunebrana and Globarzewskii each. The successful cross-amplified loci can be used for research on population genetics and gene flow of the three target species.  相似文献   

3.
Accurate species-level identifications underpin many aspects of basic and applied biology;however,identifications can be hampered by a lack of discriminating morphological characters,taxonomic expertise or time.Molecular approaches,such as DNA"barcoding"of the cytochrome c oxidase(COI)gene,are argued to overcome these issues.However,nuclear encoding of mitochondrial genes(numts)and poor amplification success of suboptimally preserved specimens can lead to erroneous identifications.One insect group for which these molecular and morphological problems are significant are the dacine fruit flies(Diptera:Tephritidae:Dacini).We addressed these issues associated with COI barcoding in the dacines by first assessing several"universal"COI primers against public mitochondrial genome and numt sequences for dacine taxa.We then modified a set of four primers that more closely matched true dacine COI sequence and amplified two overlapping portions of the COI barcode region.Our new primers were tested alongside universal primers on a selection of dacine species,including both fresh preserved and decades-old dry specimens.Additionally,Bactrocera tiyoni mitochondrial and nuclear genomes were compared to identify putative numts.Four numt clades were identified,three of which were amplified using existing universal primers.In contrast,our new primers preferentially amplified the"true"mitochondrial COI barcode in all dacine species tested.The new primers also successfully amplified partial barcodes from dry specimens for which full length barcodes were unobtainable.Thus we recommend these new primers be incorporated into the suites of primers used by diagnosticians and quarantine labs for the accurate identification of dacine species.  相似文献   

4.
Stacked wooden fruit bins are frequent overwintering sites for overwintering diapausing codling moth larvae. Control strategies against the codling moth (Cydia pomonella) (Lepidoptera: Tortricidae) in South Africa have been hampered by the reinfestation of orchards from nearby stacked infested fruit bins and by the movement of infested bins between orchards. Worldwide, wooden fruit bins are systematically being replaced with plastic bins, however in South Africa this will not be accomplished in the near future. The objective of this study was to evaluate the potential of two recycled commercially available entomopathogenic nematode (EPN) species, Heterorhabditis bacteriophora and Steinernema feltiae, as well as of a local species, Steinernema yirgalemense, to disinfest miniature wooden fruit bins under controlled conditions in the laboratory. After dipping miniature bins loaded with codling moth larvae in a suspension of 25?IJs/mL of each of the three EPN species, under optimum conditions of temperature and humidity, the highest percentage of control was obtained using S. feltiae (75%). The addition of adjuvants significantly increased S. feltiae infectivity to >95%, whereas it did not result in a significant increase in H. bacteriophora or S. yirgalemense infectivity.  相似文献   

5.
Aoshima  M.  Naka  H.  Tsuchida  K. 《Applied Entomology and Zoology》2020,55(2):231-240

Mating disruption using synthetic sex pheromones is an effective pest-control method to prevent the evolution of insecticide resistance. Although the sex pheromone of the yellow peach moth Conogethes punctiferalis (Guenée) (Lepidoptera, Crambidae) has already been identified and artificially synthesized, mating disruption in the field has been less successful than expected. Such ineffectiveness may be caused by genetic variation in the attractiveness of the sex pheromone across moth populations. To evaluate this possibility, we evaluated the genetic population structure of this species across 10 populations in Japan. We found that two clades were present at the mtDNA COI region, each clade was sympatrically distributed, and no genetic structure was detected among the populations. In addition, the frequencies of the two clades did not differ between the samples in pheromone traps using two types of blend and those hand collected from infested fruit. Our mating experiment revealed random mating between the two clades. Our results strongly suggest that the genetic variation in the attractiveness of the sex pheromone is minimal and symmetrical gene flow likely occurs between the clades in the field.

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6.
Sex pheromone mating disruption (MD) is an approach used to control several moth pest species of pome fruit by disrupting the ability of the males to find females and consequently prevent mating. The following experiments were performed to determine the effectiveness of several new and experimental sex pheromone MD technologies, and dispenser densities for simultaneous control of the codling moth (CM), Cydia pomonella (L.), and the oriental fruit moth (OFM), Grapholita molesta (Busck) (both Lepidoptera: Tortricidae), in Pennsylvania apple orchards. In one study, three MD approaches to control CM and oriental fruit moth – CM and OFM Disrupt Micro‐Flakes, Isomate CM/OFM TT, and both a CideTrak OFM and a CideTrak CM dispenser containing both codlemone and pear ester – and an insecticides‐only treatment were compared over the course of 2 years. In the other studies, the efficacy of several CheckMate Duel dispenser densities (i.e., 250, 375, 425, and 500 dispensers ha?1) were compared against Isomate CM/OFM TT, and an insecticides‐only treatment. The CideTrak CM/pear ester combination and Isomate CM/OFM TT treatments both substantially reduced CM captures in traps in 2007 and 2008. Meanwhile, OFM trap shutdown was highest in the CheckMate Duel densities of 375 (99.9 ± 0.08%) and 500 dispensers ha?1 (98.9 ± 0.07%) and the Isomate CM/OFM TT treatment (98.0 ± 1.13%), and lowest in the 250 dispensers ha?1 density treatment (94.3 ± 3.23%). In orchards where OFM is the dominant pest species, a CheckMate Duel dispenser density of 375 ha?1 is necessary for effective control, whereas higher densities are needed to control CM.  相似文献   

7.
Although several molecular diagnostic techniques are available for the identification of the apple‐feeding pests Grapholita molesta Busck and Grapholita dimorpha Komai, these pests are severely affecting apple orchards in Korea. These two pests may be misidentified or the available molecular diagnostic techniques may not facilitate the simultaneous identification of the morphological features of both species. In this study, we developed a multiplex assay for these two species using the polymerase chain reaction – restriction fragment length polymorphism (PCR‐RFLP) method. Sixty‐two specimens were collected from apples presumed infested with moth larvae and from pheromone traps from 2013 to 2014. Both species were identified morphologically, and a partial region of the cytochrome b gene was sequenced to design primers for PCR‐RFLP. Digestion profiles of G. molesta and G. dimorpha, using the Sau3A1 restriction enzyme, were characterized using three DNA fragments each for G. molesta (363 bp, 91 bp and 31 bp) and G. dimorpha (220 bp, 234 bp and 31 bp). The RFLP assay developed for both species in this study was more efficient and accurate than other currently used diagnostic assays and would be helpful to identify field‐collected specimens for pest control research.  相似文献   

8.
Two molecular identification techniques for differentiating six lepidopteran pests infesting apples in Korea are presented. These six species include two internal fruit feeders (Grapholita molesta and Carposina sasakii), two leaf rollers (Adoxophyes sp. and Archips breviplicanus) and two leaf miners (Phyllonorycter ringoniella and Lyonetia prunifoliella). All species occur until near harvest and reduce apple production. A 489 bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in these six species. The sequence was used to select species-specific restriction enzyme sites and to design diagnostic polymerase chain reaction (PCR) primers, resulting in the development of restriction fragment length polymorphism (RFLP)-PCR and diagnostic PCR. These methods were reliable and rapid in the identification of these six species.  相似文献   

9.
The pyralid moth Eldana saccharina Walker is an indigenous insect widely distributed throughout sub-Saharan Africa. Studies have shown that populations from West Africa have distinct behavioural differences compared to populations from East and southern Africa. In addition, the parasitoid guilds attacking populations in these different regions are markedly different. This marked geographical variation evoked a hypothesis of genetic differentiation. To evaluate this hypothesis a molecular analysis was conducted on populations of E. saccharina from throughout much of the species’ range, using the cytochrome c oxidase subunit I (COI) region of the mitochondrial genome. A minimum spanning network and a maximum parsimony tree separated the 21 specimens into three distinct groups. Results revealed the presence of substantial genetic differentiation that is related to geographic variation.  相似文献   

10.
Experiments were conducted in North and South America during 2012–2013 to evaluate the use of lure combinations of sex pheromones (PH), host plant volatiles (HPVs) and food baits in traps to capture the oriental fruit moth, Grapholita molesta (Busck), and codling moth, Cydia pomonella (L.), in pome and stone fruit orchards treated with sex pheromones. The combination of the sex pheromone of both species (PH combo lure) significantly increased G. molesta and marginally decreased C. pomonella captures as compared with captures of each species with either of their sex pheromones alone. The addition of a HPV combination lure [(E,Z)‐2,4‐ethyl decadienoate plus (E)‐β‐ocimene] or acetic acid used alone or together did not significantly increase the catch of either species in traps with the PH combo lure. The Ajar trap baited with terpinyl acetate and brown sugar (TAS bait) caught significantly more G. molesta than the delta trap baited with PH combo plus acetic acid in California during 2012. The addition of a PH combo lure to an Ajar trap significantly increased catches of G. molesta compared to the use of the TAS bait or PH combo lure alone in 2013. Female G. molesta were caught in TAS‐baited Ajar traps at similar levels with or without the use of additional lures. Ajar traps baited with the TAS bait alone or with (E)‐β‐ocimene and/or PH combo lures caught significantly fewer C. pomonella than delta traps with sex pheromone alone. Ajar traps with 6.4‐mm screened flaps caught similar numbers of total and female G. molesta as similarly baited open Ajar traps, and with a significant reduction in the catch of non‐targets. Broader testing of HPV and PH combo lures for G. molesta in either delta or screened or open Ajar traps is warranted.  相似文献   

11.
A polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP)-based method for species identification was applied to seven Japanese Lymantria species, including four Asian gypsy moth (AGM) species. We sequenced the partial end of the cytochrome c oxidase I (COI) gene, tRNA leucine, COII gene, and partial end of the tRNA lysine in mitochondrial DNA (mtDNA) for one individual of each of the seven species. We analyzed the recognition sites of three restriction endonucleases and constructed a scheme for Lymantria species identification using PCR–RFLP. We then applied the scheme to 291 individuals from 45 populations of seven species. We found that all seven species were correctly identified using PCR–RFLP. These results suggest that PCR–RFLP is useful for identifying Japanese Lymantria species, which may be detected at Japanese ports.  相似文献   

12.
Male and female moth catches of Grapholita molesta (Busck) in traps were evaluated in stone and pome fruit orchards untreated or treated with sex pheromones for mating disruption in Uruguay, Argentina, Chile, USA, and Italy from 2015 to 2017. Trials evaluated various blends loaded into either membrane cup lures or septa. Membrane lures were loaded with terpinyl acetate (TA), acetic acid (AA) and (Z)‐3‐hexenyl acetate alone or in combinations. Two septa lures were loaded with either the three‐component sex pheromone blend for G. molesta alone or in combination with codlemone (2‐PH), the sex pheromone of Cydia pomonella (L). A third septum lure included the combination sex pheromone blend plus pear ester, (E,Z)‐2,4‐ethyl decadienoate (2‐PH/PE), and a fourth septum was loaded with only β‐ocimene. Results were consistent across geographical areas showing that the addition of β‐ocimene or (Z)‐3‐hexenyl acetate did not increase moth catches. The addition of pear ester to the sex pheromone lure marginally increased moth catches. The use of TA and AA together significantly increased moth catches compared with the use of only one of the two components. Traps with the TA/AA lure outperformed the Ajar trap baited with a liquid TA plus sugar bait. The emission rate of AA was not a significant factor affecting the performance of the TA/AA lure. The addition of TA/AA significantly increased moth catches when combined with the 2‐PH lure. The TA/AA lure also allowed traps to catch both sexes. Catch of C. pomonella with the 2‐PH lure was comparable to the use of codlemone; however, moth catch was significantly reduced with the 2‐PH/PE lure. Optimization of these complex lures can likely further improve managers’ ability to monitor G. molesta and help to develop multispecies tortricid lures for use in individual traps.  相似文献   

13.

Lytopylus rufipes (Nees) (Hymenoptera: Braconidae: Agathidinae) is a potential natural enemy of the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), but there is no established method to rear this wasp continuously. In the laboratory, female wasps can produce both female and male progenies without mating (deuterotokous), but host-infested plants are necessary to trigger oviposition behavior. In this study, immature apples were used because they keep well. Grapholita molesta larvae were transferred to immature apples, and then exposed to L. rufipes females. After parasitization, these apples were transferred to blocks of artificial diet (Silkmate 2M) for further rearing. Using this transitional diet system, L. rufipes females develop in 25.1?±?1.8 (mean?±?SD) days from egg to adult, but male wasps require only 23.8?±?1.0 days. Furthermore, the longevity of female wasps was 12.2?±?7.3 (mean?±?SD) days, and the parasitism rate was 26.4 (95% Cl: 22.2–30.6). Female wasps can attack host larvae in all instars, but parasitism of first-instar larvae is more successful. Therefore, first-instar larvae of G. molesta are recommended for establishing a L. rufipes colony under laboratory conditions. This system decreases the requirement for plant material and maintains continuous production of L. rufipes.

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14.
Combined attracticide formulations targeting Oriental fruit moth, Grapholita molesta (Busck), and codling moth, Cydia pomonella (L.), were tested in a field trapping experiment. Capture of male codling moths in traps baited with the combined formulation was reduced compared with traps baited with the codling moth formulation alone, whereas capture of male Oriental fruit moth was increased compared with traps baited with the Oriental fruit moth formulation alone. Subsequent wind tunnel experiments showed that a single locus of the mixed attracticide formulation or close parallel presentation of the two formulations enhanced source contact by male Oriental fruit moths but did not influence earlier behaviors. However, the two formulations presented in a serial arrangement to Oriental fruit moth males in the wind tunnel resulted in enhanced lock-on, upwind flight, and source contact behaviors. In addition, male Oriental fruit moths remained on mixed pheromone droplets of the paste matrix longer than on droplets of the Oriental fruit moth formulation alone. The increased time spent on the mixed droplet was correlated with a more rapid poisoning and a greater proportion of poisoned males compared with males exposed to the Oriental fruit moth attracticide alone. These results demonstrate that a combined attracticide formulation will have different effects on each of the targeted species. It is anticipated that, due to decreased attractiveness, a combined formulation would be less effective against the codling moth. However, a mixed formulation, due to increased attractiveness and toxicity, could be more effective against the Oriental fruit moth under field conditions.  相似文献   

15.
Studies were conducted in Chile and the United States to compare the attractiveness of various commercial sex pheromone lures and two experimental lures for oriental fruit moth, Grapholita molesta (Busck), in peach orchards treated with or without sex pheromone dispensers. The experimental lures contained the three‐component sex pheromone blend of G. molesta: Z‐8‐dodecenyl acetate, E‐8‐dodecenyl acetate and Z‐8‐dodecenol (Z8‐12:OH), and the sex pheromone of codling moth, Cydia pomonella (L.), (E,E)‐8,10‐dodecadien‐1‐ol, (codlemone). Commercial lures varied in their substrate, initial loading and blend ratio of components. Significant differences in male catches were found among commercial lures in orchards treated with or without sex pheromone dispensers. Experimental lures with the addition of codlemone significantly increased the catches of G. molesta using lures loaded with 0%, 1% or 5% Z8‐12:OH in the G. molesta blend compared with the same ratio of components in just the G. molesta blend. The experimental lures were significantly more attractive than all commercial lures in the untreated orchard. However, moth catch with the experimental lures in the sex pheromone‐treated orchard was only intermediate among all of the lures tested. These findings highlight the need to develop more effective and standardized lures that can be used in trap‐based monitoring programme for this important pest.  相似文献   

16.
Studies compared moth captures of three pests (Lepidoptera: Tortricidae) of apple, Malus domestica Borkhausen, in delta traps using removable liners coated with either a sticky gel (SG) or a hot‐melt pressure sensitive (HMPS) adhesive. Laboratory and field studies with Cydia pomonella (L.), Choristoneura rosaceana (Harris) and Grapholita molesta (Busck) demonstrated that traps with either liner, baited with a pair of virgin females catch males, but at significantly different levels. In the field, male moth captures in traps with the HMPS liner were significantly greater than in traps with the SG liner for C. rosaceana, and G. molesta; but not for C. pomonella. Similar results were observed in laboratory studies using flight tunnels. Additional studies demonstrated that this difference in moth captures between liners was not due to levels of female mortality, but instead was correlated with the occurrence of the female's ventral abdominal surface becoming stuck in the adhesive. Studies showed that a significantly greater proportion of females of all species had their ventral abdomen stuck in the SG than HPMS adhesive on day 1, but only G. molesta and C. rosaceana on day 3. In addition, the tackiness of the two adhesives affected moth movement on the liner with males and females of all species moving farther on liners with SG than HMPS adhesive. A greater proportion of female G. molesta and C. pomonella were either stuck supined or laterally on the SG than HMPS adhesive, and females in this position captured as many males as when prone and unstuck on the liner. Our studies demonstrate that adhesives can secondarily influence male moth captures on trap liners when used with bisexual attractants, and that adhesive type should be considered when developing action thresholds.  相似文献   

17.
The phenology of oriental fruit moth, Grapholita molesta (Busck), on apple (Malus spp.) in North Carolina was studied using pheromone traps and egg sampling in abandoned and commercial orchards in 2000 and 2001, with subsequent development of an oviposition degree-day model and management studies in relation to codling moth, Cydia pomonella (L.), phenology. Oriental fruit moth eggs were found in greater numbers on leaves early and on fruit later in the growing season, on the top versus the bottom of the leaf surface, and on the calyx area versus the side or stem end of the fruit. A degree-day (DD) model to predict oriental fruit moth oviposition was developed based on temperature accumulations from peak moth trap capture of the first (overwintering) generation, by using 7.2 and 32.2 degrees C as the temperature limits. The model predicted four ovipositing generations of oriental fruit moth with the second beginning 507 DD after peak moth catch. Using predictions of the oriental fruit moth and codling moth degree-day oviposition models, an experiment was conducted to determine the level of second generation oriental fruit moth control with methoxyfenozide applied under different scenarios for first generation codling moth. Methoxyfenozide was equally effective in managing codling moth and oriental fruit moth for all treatment timings.  相似文献   

18.
The codling moth, Cydia pomonella (L.), and oriental fruit moth, Grapholita molesta (Busck), are two key pests of apple (Malus domestica Borkh.) in North Carolina. Growers extensively relied on organophosphate insecticides, primarily azinphosmethyl, for > 40 yr to manage these pests. Because of organophosphate resistance development and regulatory actions, growers are transitioning to management programs that use new, reduced-risk, and OP-replacement insecticides. This study evaluated the toxicity of a diversity of replacement insecticides to eggs, larvae, and adults, as well as an assessment of their residual activity, to codling moth and oriental fruit moth. Laboratory-susceptible strains of both species were used for all bioassays. Fresh field-harvested apples were used as a media for assessing the ovicidal activity of insecticides. For larval studies, insecticides were topically applied to the surface of lima bean-based diet, onto which neonates were placed. Toxicity was based on two measures of mortality; 5-d mortality and development to adult stage. Ovicidal bioassays showed that oriental fruit moth eggs were generally more tolerant than codling moth eggs to insecticides, with novaluron, acetamiprid, and azinphoshmethyl having the highest levels of toxicity to eggs of both species. In contrast, codling moth larvae generally were more tolerant than oriental fruit moth to most insecticides. Methoxyfenozide and pyriproxyfen were the only insecticides with lower LC50 values against codling moth than oriental fruit moth neonates. Moreover, a number of insecticides, particularly the IGRs methoxyfenozide and novaluron, the anthranilic diamide chlorantriliprole, and the spinosyn spinetoram, provided equal or longer residual activity against codling moth compared with azinphosmethyl in field studies. Results are discussed in relation to their use in devising field use patterns of insecticides and for insecticide resistance monitoring programs.  相似文献   

19.
A series of studies were conducted to examine the residual activity and toxicity of the ecdysone agonists tebufenozide and methoxyfenozide to codling moth, Cydia pomonella (L.), and oriental fruit moth, Grapholita molesta (Busck), in North Carolina apple systems. Methoxyfenozide exhibited greater activity than tebufenozide against codling moth eggs in dose-response bioassays, with a 4.5- and 5.3-fold lower LC50 value to eggs laid on fruit treated before or after oviposition, respectively. Oriental fruit moth eggs were 57- and 12-fold less sensitive to methoxyfenozide than were codling moth eggs on fruit treated before and after oviposition, respectively. Methoxyfenozide was effective in reducing larval entries of both codling moth and oriental fruit moth in field residual activity bioassays, exhibiting activity for at least 28 d after application. Residue breakdown on fruit was approximately 80% at 28 d after treatment for both methoxyfenozide and tebufenozide, with the most rapid residue decline (60%) occurring during the first 14 d after application. Two applications of methoxyfenozide applied at 14-d intervals provided better canopy coverage and higher residue levels than one application. Spray volume (683 versus 2,057 liters/ha) did not affect the efficacy of methoxyfenozide. Leaf and fruit expansion during the season was measured to determine potential plant-growth dilution effects on residual activity. There was very little increase in leaf area after mid May, but increase in fruit surface area over the season was described by a second order polynomial regression. Implications for codling moth and oriental fruit moth management programs are discussed.  相似文献   

20.
The ability of a sufficient number of individuals to disperse is crucial for long‐term survival of populations. However, dispersal is often energetically costly, and thus is expected to trade‐off against other life‐history traits. In insect pest species, the occurrence of individuals with high flight activity challenges management practices. We performed artificial selection on flight activity and measured correlated responses to selection in the oriental fruit moth, Grapholita (= Cydia) molesta, a widely distributed and expanding lepidopteran pest of fruit crops. Both sexes rapidly responded to the imposed regime of divergent selection, indicating an adaptive potential of flight activity in this species. Upward‐selected moths died sooner than downward‐selected ones, providing evidence for a cost of flight activity to adult survival, reputedly associated with enhanced metabolic rates. Oppositely‐selected females had similar total reproductive output, disproving a trade‐off between dispersal and reproduction, although females with higher flight activity laid their eggs sooner. The ratio of body weight to forewing surface (forewing loading) did not significantly differ between selected lines. The present study contributes to the understanding of dispersal evolution, and also provides new insights into life‐history theory as well as important baseline data for the improvement of pest management practices. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 879–889.  相似文献   

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