Patterns of mitochondrial haplotype diversity in the invasive pest Epiphyas postvittana (Lepidoptera: Tortricidae)

The light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), is a horticultural pest of Australia and New Zealand that has more recently invaded Hawaii, Europe, and California. A 2,216-bp region of the mitochondrial genome containing the cytochrome oxidase I and II genes was sequenced from 752 individuals. Haplotype network analyses revealed a major split between a predominantly Western Australian clade and all other samples, suggestive of either a deep genetic divergence or a cryptic species. Nucleotide and haplotype diversity were highest in the country of origin, Australia, and in New Zealand populations, with evidence of haplotype sharing between New Zealand and Tasmania. Nucleotide and haplotype diversity were higher in California than within the British Isles or Hawaii. From the total of 96 haplotypes, seven were found in California, of which four were private. Within California, there have been at least two introductions; based on genetic diversity we were unable to assign a likely source for a single moth found and eradicated in Los Angeles in 2007; however, our data suggest it is unlikely that Hawaii and the British Isles are sources of the major E. postvittana population found throughout the rest of the state since 2006.     

A chemically defined medium for rearing Epiphyas postvittana (Lepidoptera: Tortricidae)

Abstract

The endemic New Zealand genus Neolimnia is redescribed, and a key is given to the 14 species now included. Four new species are described - Neolimnia raiti, N. pepekeiti, N. ura, and N. repo. Two new synonymies are proposed; N. ocellata Tonnoir &; Malloch with N. castanea (Hutton); and N. dubiosa Tonnoir &; Malloch with N. irrorata Tonnoir &; Malloch. Lectotypes are designated for N. tranquilla (Hutton), N. castanea (Hutton), N. obscura (Hutton), and N. striata (Hutton). Male postabdomens of all species are illustrated. Character variation and generic, subgeneric, and species relationships are discussed briefly.     

Pre- and postharvest effects of lufenuron on Epiphyas postvittana (Lepidoptera: Tortricidae)

First-, third-, and fifth-instar Epiphyas postvittana (Walker) were exposed to a range of lufenuron concentrations (0-200 ppm) incorporated into synthetic diet and their subsequent development and mortality responses were determined. For all instars the greatest change in mortality response occurred over lufenuron concentrations < or = 3 ppm. However, third and fifth instars displayed an increase in mortality earlier than first instars, and were more sensitive to the lower lufenuron concentrations in this range. Only first and third instars subjected to < or = 2.5 ppm lufenuron survived the 26-d exposure trial. No larvae first exposed to lufenuron as first or third instars survived to pupation if ingesting concentrations of > or = 1 and > or = 3 ppm, respectively. Consumption of lower lufenuron concentrations by these larvae delayed pupation and resulted in pupal deformity. In contrast, fifth instars subjected to 100 ppm were capable of surviving the 26-d trial period and displayed a slower progressive reduction in survival to pupation with increase in lufenuron concentration. Also in contrast to more immature stages, fifth instars exposed to lufenuron developed more rapidly to pupation than larvae not exposed to the insect growth regulator (IGR), and all resulting pupae were normal. Third instars were exposed to sublethal lufenuron concentrations (0-3 ppm) for 4 d and the fourth-instar survivors subjected to a controlled atmosphere cold storage treatment (2% O2, 2% CO2, 0.6 degree C). Larvae ingesting diet containing 0.5 ppm (and to a lesser extent 1 ppm) lufenuron required longer exposure to the postharvest treatment to achieve > or = 95% mortality than larvae not ingesting the IGR. However, the analogous mortality response of larvae exposed to 3 ppm lufenuron was comparable to the control.     

Aseptic rearing of Epiphyas postvittana (Lepidoptera: Tortricidae) on a meridic diet

The light brown apple moth, Epiphyas postvittana (Walker), was reared aseptically from newly hatched larvae to adults for three generations on a meridic diet. Growth and development on this diet were comparable with that on oligidic diets and on host‐plants. At 25 ± 1°C, 50–60% R.H., and 12‐h photophase, the average larval period was approx. 28 days, pre‐pupal and pupal 11–12 days, and pre‐oviposition approx. 2 days; the incuva‐tion period of eggs was approx. 7 days. The average developmental period from egg to adult was thus about 49 days; 89 % of larvae reached the adult stage. The average fecundity was 598 eggs per female, and the sex ratio was nearly 1:1. There were no differences in the rate of development or in fecundity of insects reared on media with and without a mould inhibitor.     

Light brown apple moth (Epiphyas postvittana) (Lepidoptera: Tortricidae) colonization of California

In response to the confirmed detection of the light brown apple moth, Epiphyas postvittana, in California, approximately 53,000 pheromone-baited Jackson traps were deployed and more than 246,000 males were caught (February 2007–February 2010). Approximately 46,000 manually entered catch records were corrected for errors and converted into catch per trap per day. As empty trap data (zeros) were not recorded, we added zeros between first and last catch for each trap based on the stated servicing period (~30,000), before analyzing for trends. Residual data error rate was estimated as 1.5 %. San Francisco and Santa Cruz counties had relatively high trap catches immediately upon trap set, and remained the leading population centers, while most other counties showed a more general trend of a slow build-up in catch over time (12 counties). An exponential increase in trap catch was observed in four counties with sufficient data. The pattern of spread indicated natural, as well as anthropogenic-assisted spread rates, with populations appearing well ahead of the invasion front. This jump dispersal is probably due to movement of host plants, unsurprising since eggs of this polyphagous moth are readily laid on foliage. There was evidence of seasonality in spread, probably linked to the phenology of the insect. There was a positive relationship between catch and known host tree preference, suggesting that trap placement in preferred hosts could add sensitivity to future surveys. Recommendations include the improved provision of data acquisition by telecommunications, standardization of data input, more archiving, and frequent analysis of trap catches. The rapid rate of population growth demonstrated in two counties and spread across many others supports the hypothesis of the recent arrival of E. postvittana in California.     

Development of Epiphyas postvittana (Lepidoptera: Tortricidae) on leaves and fruit of orange trees

Development of Epipyas postvittna (Walker) (Lepidoptera: Tortricidae), on leaves and fruit of 'Valencia', 'Washington navel', and 'Navelina' orange trees was studied under constant and fluctuating temperatures. E. postvittna was able to complete its life cycle feeding exclusively on leaves or fruit of orange trees. However, larval survival rate was very low (< 20%) on orange tissues compared with that on noncitrus hosts. Among the four types of orange tissues, young orange leaves and fruit afforded larvae higher survival rates than mature orange leaves and fruit. Fruit (young or mature) produced heavier pupae than leaves (young or mature). Larvae developed more slowly on mature orange fruit than on other orange materials and more slowly on orange leaves than on leaves of most noncitrus hosts. Degree-day accumulations based on the fastest developmental rates obtained in this study suggested that E. postvittna is capable of completing 4.4-4.7 generations per year in orange orchards in the Riverina region of New South Wales, Australia. Implications of the results in the management of the insect in citrus are discussed.     

A multiplex real-time polymerase chain reaction assay to diagnose Epiphyas postvittana (Lepidoptera: Tortricidae)

A molecular assay for diagnosis of light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), in North America is reported. The assay multiplexes two TaqMan real-time polymerase chain reaction (RT-PCR) probe systems that are designed to target DNA segments of the internal transcribed spacer region 2 (ITS2) and 18S rRNA gene. The RT-PCR probe designed for the 18S target recognizes a DNA sequence conserved in all of the moths included in the study and functions as a control in the assay. The second probe recognizes a segment of the ITS2 specifically found in E. postvittana and not found in the other moths included in the study, i.e., this segment is not conserved. Inclusion of the two markers in a single multiplex reaction did not affect assay performance. The assay was tested against 637 moths representing > 90 taxa in 15 tribes in all three subfamilies in the Tortricidae. The assay generated no false negatives based on analysis of 355 E. postvittana collected from California, Hawaii, England, New Zealand, and Australia. Analysis of a data set including 282 moths representing 41 genera generated no false positives. Only three inconclusive results were generated from the 637 samples. Spike experiments demonstrated that DNA contamination in the assay can affect samples differently. Contaminated samples analyzed with the ITS2 RT-PCR assay and DNA barcode methodology by using the cytochrome oxidase I gene can generate contradictory diagnoses.     

Pheromone binding proteins of Epiphyas postvittana (Lepidoptera: Tortricidae) are encoded at a single locus

The light brown apple moth, Epiphyas postvittana (Tortricidae: Lepidoptera) uses a blend of (E)-11-tetradecenyl acetate and (E,E)-9,11-tetradecadienyl acetate as its sex pheromone. Odorant binding proteins, abundant in the antennae of male and female E. postvittana, were separated by native PAGE to reveal four major proteins with distinct mobilities. Microsequencing of their N-terminal residues showed that two were general odorant binding proteins (GOBPs) while two were pheromone binding proteins (PBPs). Full length cDNAs encoding these proteins were amplified using a combination of PCR and RACE-PCR. Sequence of the GOBPs revealed two genes (EposGOBP1, EposGOBP2), similar to orthologues in other species of Lepidoptera. Eleven cDNAs of the PBP gene were amplified, cloned and sequenced revealing two major phylogenetic clusters of PBP sequences differing by six amino acid substitutions. The position of the six amino acid differences on the protein was predicted by mapping onto the three-dimensional structure of PBP of Bombyx mori. All six substitutions were predicted to fall on the outside of the protein away from the inner pheromone binding pocket. One substitution does fall close to the putative dimerisation region of the protein (Ser63Thr). Expression of three of the cDNAs in a baculovirus expression system revealed that one class encodes an electrophoretically slow form (EposPBP1-12) while the other encodes a fast form (EposPBP1-2, EposPBP1-3). A native Western of these expressed proteins compared with antennal protein extracts demonstrated that PBP is also expressed in female antennae and that PBP may be present as a dimer as well as a monomer in E. postvittana. The fast and slow forms of EposPBP1 are allelic. Westerns on single antennal pair protein extracts and allele-specific PCR from genomic DNA both show a segregating pattern of inheritance in laboratory and wild populations. Radio labelled (E)-11-tetradecenyl acetate binds to both fast and slow PBP forms in gel assays. Taken together, the genetic and biochemical data do not support the hypothesis that these PBPs are specific for each component of the E. postvittana pheromone. However, duplication of this PBP locus in the future might allow such diversification to evolve, as observed in the other species.     

Multiple mating and its effect on the reproductive success of female Epiphyas postvittana (Lepidoptera: Tortricidae)

Abstract.

• 1 Multiple mating and its effect on reproductive performance of female Epiphyas postvittana (Walker) moths were studied under controlled conditions.
• 2 The age at which the moths mated for the first time ranged from the first to the tenth day after emergence, but 71% of first matings were during the first 3 days.
• 3 The majority (63%) of females had one or two spermatophores in the bursa copulatrix. Some (24%) were found with three to five spermatophores, whereas no successful mating occurred among 13% of individuals. The number of matings was partly dependent on the number of mates available to the female. Between the range of sex ratios of one male to one female and four males to one female maximal mating success occurred at the ratio of three males to one female.
• 4 Virgin females were capable of egg-laying, but mating stimulated and accelerated oviposition. Mated individuals laid twice as many eggs as unmated ones.
• 5 The level of copulatory activity did not influence the longevity of females irrespective of the number of males available to them.
• 6 Sex ratios with greater than one male to a female improved the reproductive success by marginally increasing fecundity and fertility.
• 7 It is concluded that multiple mating would enhance population growth, and is of particular benefit to populations with a preponderance of females, as is known to occur naturally in this species.

     

The hymenopteran parasitoids of light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae) in Australia

Abstract  Epiphyas postvittana (Walker) is a serious pest of a number of horticultural crops including grapes in Australia and New Zealand. This study brings together information on the parasitoid complex associated with E. postvittana that previously was fragmented and largely inaccessible. We include species reared during a 3-year study of the parasitoids of E. postvittana in the vineyards of the Coonawarra region, South Australia, material from several Australian agricultural insect collections and records from the literature. An illustrated key is presented for 25 species of parasitoids and hyperparasitoids associated with E. postvittana , along with information on the taxonomy, identification, distribution and biology of each species. Taxa newly recorded from this host are Perilampus sp. (Perilampidae), and six species of Ichneumonidae: Euceros sp., Labium sp., Netelia sp., Plectochorus sp., Temalucha minuta (Morley) and Eriborus epiphyas sp. n., the latter species being described in full.     

Irradiation of adult Epiphyas postvittana (Lepidoptera: Tortricidae): egg sterility in parental and F1 generations

Adult Epiphyas postvittana Walker were irradiated using a Cobalt 60 source to determine the dose needed to achieve complete egg sterility of mated female moths, and egg sterility of female moths mated to F1 generation males. Adult male and female E. postvittana were irradiated at 100, 200, 250, and 300 Gy and their fertility (when crossed with normal moths) was compared with nonirradiated moths. Viable progeny (determined by egg hatch) were found at doses of 100 and 200 Gy, but very little at 250 and 300 Gy. In particular, there was no survival of female progeny into the F1 generation. Males irradiated at 250 and 300 Gy had very low egg eclosion rates (2.25 and 1.86% at 250 and 300 Gy, respectively) when mated with normal females. The F2 generation from those male progeny had a mean percent hatched of < 1.02%. Based on our results, a dose of 250-300 Gy is recommended for irradiation of E. postvittana adults used for sterile insect technique (SIT) if sterility of parental moths is the desired outcome. Our data also suggests that inclusion of F1 hybrid sterility rather than parental generation sterility into programs using the SIT may allow for doses lower than what we have reported, especially during initial phases of an eradication program where increase fitness of moths might be desirable. Further research is needed to verify the use of F1 hybrid sterility in light brown apple moth SIT programs.     

Modeling the potential global distribution of light brown apple moth Epiphyas postvittana (Lepidoptera: Tortricidae) using CLIMEX

Light brown apple moth Epiphyas postvittana is a significant horticultural pest native to Australia that currently has a limited global distribution. However, this pest can tolerate very heterogeneous climates and has a wide host range. It has recently established in California with considerable consequences for US international and domestic trade. It has resulted in increasing calls for targeted risk assessment so that appropriate quarantine measures can be put in place to prevent its entry into new regions and further spread. Potential global distribution has been predicted by comparing the climatic conditions of its native (Australia) and long-established (New Zealand) ranges to the rest of the world using CLIMEX. It was suggested that E. postvittana has potential to establish mainly in countries in Central and South America, southern Africa, Western Europe and Southeast Asia. The study provides basic information for further assessment of the establishment capacity of this species in new habitats, and adds to the knowledge required to make science-based decisions in biosecurity.     

Effect of temperature and treatment conditions on the mortality of Epiphyas postvittana (Lepidoptera: Tortricidae) exposed to ethanol

Morality responses were determined for 5th-instar lightbrown apple moth, Epiphyas postvittana (Walker), in the presence and absence of apples to immersion in ethanol solutions and exposure to ethanol vapor at a range of concentrations, treatment times, and temperatures. Ethanol may have caused an initial knock-down effect in E. postvittana larvae because there was a trend for larval mortality to reduce with increased periods between treatment and assessment time when immersed at 20 degrees C in 30 or 50% ethanol solutions. Mortality for larvae immersed on apples in a range of ethanol concentrations was higher than for larvae in the absence of apples. Increasing treatment temperature from 20 to 45 degrees C during ethanol immersion significantly increased larval mortality. During ethanol vapor exposure, longer treatment times were required to achieve 99% E. postvittana mortality for larvae on apples compared with those in the absence of apples.     

Morphogenetic and physiological effects of a juvenile hormone analogue on the light brown apple moth,Epiphyas postvittana (Lepidoptera: Tortricidae)

Abstract

The juvenile hormone analogue ZR-619—ethyl 11-methoxy-3,7,11-trimethyl-(2E,4E-dodecadienethiolate) —produced morphological and physiological effects when fed via artificial diet to larvae of Epiphyas postvittana (Walker). Morphological effects included changes in larval head and antennal structures after instar V (supernumerary instars); deformation of pupal and adult structures, particularly in genitalia and wings; and development of individuals with varying mixtures of larval and pupal structures. Physiological effects included prolongation of larval life, increase in larval weight, increase in larval instars, and decrease in fecundity. Effects were directly related to dosage concentrations.     

Host Plants Affect the Foraging Success of Two Parasitoids that Attack Light Brown Apple Moth Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae)

The light brown apple moth, Epiphyas postvittana is a key pest of wine grapes in Australia. Two parasitoids, Dolichogenidea tasmanica and Therophilus unimaculatus, attack the larval stage of this pest. D. tasmanica is dominant in vineyards, whereas T. unimaculatus is mainly active in native vegetation. We sought to understand why they differ in their use of habitats. Plants are a major component of habitats of parasitoids, and herbivore-infested plants influence parasitoid foraging efficiency by their architecture and emission of volatile chemicals. We investigated how different plant species infested by E. postvittana could affect the foraging success of the two parasitoid species in both laboratory and field experiments. Four common host-plant species were selected for this study. In paired-choice experiments to determine the innate foraging preferences for plants, both parasitoid species showed differences in innate search preferences among plant species. The plant preference of D. tasmanica was altered by oviposition experience with hosts that were feeding on other plant species. In a behavioral assay, the two parasitoid species allocated their times engaged in various types of behavior differently when foraging on different plant species. For both parasitoids, parasitism on Hardenbergia violacea was the highest of the four plant species. Significantly more larvae dropped from Myoporum insulare when attacked than from the other three host-plant species, which indicates that parasitism is also affected by interactions between plants and host insects. In vineyards, parasitism by D. tasmanica was significantly lower on M. insulare than on the other three host-plant species, but the parasitism rates were similar among the other three plant species. Our results indicate that plants play a role in the habitat preferences of these two parasitoid species by influencing their foraging behavior, and are likely to contribute to their distributions among habitats.     

Variations in life history traits and flight capacity among populations of the light brown apple moth Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae)

Abstract The empirical study of interpopulation variation in life history and other fitness traits has been an important approach to understanding the ecology and evolution of organisms and gaining insight into possible sources of variation. We report a quantitative analysis for variations of five life history traits (larval developmental time, adult body weight, adult lifespan, age at first reproduction, total fecundity) and flight capacity among populations of Epiphyas postvittana originating from four localities in Australia and one in New Zealand. These populations were compared at two temperatures (15° and 25°C) after being maintained under uniform laboratory conditions for 1.5 generations, so that the relative role of genetic divergence and phenotypic plasticity in determining interpopulation variation could be disentangled. Genetic differentiation between populations was shown in all measured traits, with the greatest divergence occurring in developmental time, fecundity and adult body size. However, these traits were highly sensitive to changes in environmental temperatures; and furthermore, significant interactions between population and temperature occurred in all traits except for flight capacity of female moths. Thus, phenotypic plasticity may be another cause of interpopulation variation. The interpopulation variation for some measured traits was apparently related to climatic differences found where the populations originated. Individuals of the populations from the warmer climates tended to develop more slowly at immature stages, producing smaller and less fecund moths but with stronger flight capacity, in comparison to those from the cooler regions. It seems, therefore, that natural populations of E. postvittana have evolved different strategies to cope with local environmental conditions.     

The effects of the broad-specificity lipase inhibitor, tetrahydrolipstatin, on the growth, development and survival of the larvae of Epiphyas postvittana (Walker) (Tortricidae, Lepidoptera)

The effects of the lipase inhibitor, tetrahydrolipstatin (THL), on neonate Epiphyas postvittana (Walker) (Lepidoptera, Tortricidae) larvae were investigated by feeding on control artificial diets (with and without 2% ethanol) and diets containing 2% ethanol and one of three concentrations of THL (0.011%, 0.037% and 0.11%). Small but significant reductions in growth rate, percent pupation and time to pupation were observed for larvae feeding on 2% ethanol control diet compared with standard control diet, but larger reductions in all parameters occurred with increasing THL concentration. Third instar larvae fed 0.011% THL in the diet had 40% of the midgut lipase activity in the relevant control larvae and showed up-regulation of gene expression of the gastric lipase-like family but not the pancreatic lipase-like family of midgut lipases.     

Effect of a vertical object on the sex pheromone-mediated flight and landing behaviors of the male lightbrown apple moth,Epiphyas postvittana (walker) (Lepidoptera: Tortricidae)

A small (2.5-cm-wide) vertical (10-cm-high) white object influenced the sex pheromone-mediated flight and landing behaviors of maleEpiphyas postvittana. When the vertical object was positioned on a horizontal surface to the side (3–5 cm) and upwind of a pheromone source (in the middle of the surface), the distribution of landing positions of males on the surface was different from that when the object was not present; males tended to land in positions skewed toward the side of the source that the object was on. The closer the object was positioned to the source, the greater the number of males that landed on the object (rather than on the horizontal surface). This difference in landing positions (when the object was present) corresponded with changes in the flight tracks; the tracks of males flying to the surface with an object were skewed toward the object and had higher amplitude intertrack reversal distances than the tracks of males flying to a surface without a vertical object. Positioning of a vertical object progressively upwind of the source resulted, apparently, in decreased effects on the landing (and presumably flight) behavior of males. The effect of the vertical object on the flight and landing behaviors of males corresponded largely with changes in pheromone plume structure (visualized with smoke) induced by the extra turbulence in the airflow over the source. Thus it appears that the vertical object influences the behavior of maleE. postvittana largely through the olfactory sensory modality. However, when a clear, Mylar object, in place of the white object, was placed on the surface, more males landed on the Mylar object (than did on the white object), suggesting that the vertical object may also influence the behavior of males through the visual modality.