Leaf-mining Moth Research Articles

Density effect and intraspecific competition in a leaf-mining moth on bamboo leaves

When confined to a single leaf, the larvae of most leafminers should suffer intense intraspecific competition. The survival and performance of individuals are expected to decline with increasing larval density within a leaf, showing a negative density-dependent effect. The maximum density of an Elachista Treitschke (Lepidoptera: Elachistidae) miner on Indocalamus tessellatus (Munro) Keng f. (Poaceae: Bambusoideae) under natural conditions can be greater than 40 per leaf, making this a good system for studying intraspecific competition and density effects. We counted the number of leaves with different mine densities in the field and found a clumped distribution of leafminer larvae among host leaves. The emergence rates of pupae (and last-instar larvae), adults and parasitoids were inversely correlated with larval density. Pupal weight and adult weight also showed negative relationships with larval density. However, pairwise tests showed that there might be no differences in pupal or adult weight at larval densities of 1–10 mines per leaf, indicating that density-dependent effects are more obvious at densities greater than 10 mines per leaf. Intraspecific competition could lead to stable density and consistent body size in surviving individual insects, which would help maintain a sustainable population size. The density threshold could be an indicator of the balance between intraspecific competitive detriments and conspecific aggregation benefits; however, the validity of the threshold density hypothesis requires further testing and confirmation.

CONTRIBUTION TO THE KNOWLEDGE OF PHYLLONORYCTER ISSIKII (KUMATA, 1963) (LEPIDOPTERA, GRACILLARIIDAE) IN SERBIA

Phyllonorycter issikii (Kumata, 1963) is an invasive leaf-mining moth originating from Asia. It develops on plants from the genus Tilia. The first finding of this species in Serbia was in 2013, and since then, the range of the species has spread, and we have found it at five more localities on T. cordata and T. tomentosa. Ph. issikii was studied at two of these localities. The existence of two overlapping generations was determined. No significant differences in the intensity of the attack between them were noticed. Differences existed only between the localities where the research was conducted and between the Tilia species on which Ph. issikii was found. As regards the lime species, the moth preferred T. cordata to T. tomentosa at both localities. Two species of parasitoids were reared from the collected material: Pediobius saulius (Walker, 1839) and Minotetrastichus frontalis (Nees, 1834).

Systematics of Phyllocnistis leaf-mining moths (Lepidoptera, Gracillariidae) feeding on dogwood (Cornus spp.) in Northeast Asia, with the description of three new species.

During an ongoing DNA-barcoding campaign of the leaf-mining moths that feed on woody plants in Northeast Asia, four lineages of the genus Phyllocnistis (Gracillariidae, Phyllocnistinae) were discovered on dogwood (Cornus spp): P. cornella Ermolaev, 1987 on C. controversa Hemsl. (Japan: Hokkaido) and three new species – one feeding on C. controversa, C. florida L. and C. macrophylla Wall. in Japan (Honshu, Shikoku, Kyushu), a second species on C. macrophylla in China (Yunnan) and a third on Siberian dogwood Cornus alba L. in Russia (Siberia). All these species showed differences in morphology, in the barcode region of the cytochrome c oxidase I gene and in two nuclear genes (histone H3 and 28S ribosomal RNA). No correlation was found between the deep mitochondrial splits observed and the Wolbachia infection pattern. Based on both morphological and molecular evidence, the three recently discovered lineages are described here as new species: P. indistincta Kobayashi & Triberti, sp. n. (Japan), P. saepta Kirichenko, Ohshima & Huang, sp. n. (China) and P. verae Kirichenko, Triberti & Lopez-Vaamonde, sp. n. (Russia). In addition, the authors re-describe the adult morphology of P. cornella, provide the first record of this species from Japan and highlight the diagnostic characters that allow these Cornus-feeding Phyllocnistis species to be distinguished.

Attraction of Three Mirid Predators to Tomato Infested by Both the Tomato Leaf Mining Moth Tuta absoluta and the Whitefly Bemisia tabaci.

Plants emit volatile compounds in response to insect herbivory, which may play multiple roles as defensive compounds and mediators of interactions with other plants, microorganisms and animals. Herbivore-induced plant volatiles (HIPVs) may act as indirect plant defenses by attracting natural enemies of the attacking herbivore. We report here the first evidence of the attraction of three Neotropical mirid predators (Macrolophus basicornis, Engytatus varians and Campyloneuropsis infumatus) toward plants emitting volatiles induced upon feeding by two tomato pests, the leaf miner Tuta absoluta and the phloem feeder Bemisia tabaci, in olfactometer bioassays. Subsequently, we compared the composition of volatile blends emitted by insect-infested tomato plants by collecting headspace samples and analyzing them with GC-FID and GC-MS. Egg deposition by T. absoluta did not make tomato plants more attractive to the mirid predators than uninfested tomato plants. Macrolophus basicornis is attracted to tomato plants infested with either T. absoluta larvae or by a mixture of B. tabaci eggs, nymphs and adults. Engytatus varians and C. infumatus responded to volatile blends released by tomato plants infested with T. absoluta larvae over uninfested plants. Also, multiple herbivory by T. absoluta and B. tabaci did not increase the attraction of the mirids compared to infestation with T. absoluta alone. Terpenoids represented the most important class of compounds in the volatile blends and there were significant differences between the volatile blends emitted by tomato plants in response to attack by T. absoluta, B. tabaci, or by both insects. We, therefore, conclude that all three mirids use tomato plant volatiles to find T. absoluta larvae. Multiple herbivory did neither increase, nor decrease attraction of C. infumatus, E. varians and M. basicornis. By breeding for higher rates of emission of selected terpenes, increased attractiveness of tomato plants to natural enemies may improve the effectiveness of biological control.

A statistical method for analysing cospeciation in tritrophic ecology using electrical circuit theory.

We introduce a new method to test efficiently for cospeciation in tritrophic systems. Our method utilises an analogy with electrical circuit theory to reduce higher order systems into bitrophic data sets that retain the information of the original system. We use a sophisticated permutation scheme that weights interactions between two trophic layers based on their connection to the third layer in the system. Our method has several advantages compared to the method of Mramba et al. [Mramba, L. K., S. Barber, K. Hommola, L. A. Dyer, J. S. Wilson, M. L. Forister and W. R. Gilks (2013): "Permutation tests for analyzing cospeciation in multiple phylogenies: applications in tri-trophic ecology," Stat. Appl. Genet. Mol. Biol., 12, 679-701.]. We do not require triangular interactions to connect the three phylogenetic trees and an easily interpreted p-value is obtained in one step. Another advantage of our method is the scope for generalisation to higher order systems and phylogenetic networks. The performance of our method is compared to the methods of Hommola et al. [Hommola, K., J. E. Smith, Y. Qiu and W. R. Gilks (2009): "A permutation test of host-parasite cospeciation," Mol. Biol. Evol., 26, 1457-1468.] and Mramba et al. [Mramba, L. K., S. Barber, K. Hommola, L. A. Dyer, J. S. Wilson, M. L. Forister and W. R. Gilks (2013): "Permutation tests for analyzing cospeciation in multiple phylogenies: applications in tri-trophic ecology," Stat. Appl. Genet. Mol. Biol., 12, 679-701.] at the bitrophic and tritrophic level, respectively. This was achieved by evaluating type I error and statistical power. The results show that our method produces unbiased p-values and has comparable power overall at both trophic levels. Our method was successfully applied to a dataset of leaf-mining moths, parasitoid wasps and host plants [Lopez-Vaamonde, C., H. Godfray, S. West, C. Hansson and J. Cook (2005): "The evolution of host use and unusual reproductive strategies in achrysocharoides parasitoid wasps," J. Evol. Biol., 18, 1029-1041.], at both the bitrophic and tritrophic levels.

Quantitative analysis of the causes of outbreaks in leaf-mining insects: application to the common leaf-miner Leucoptera malifoliella (Lepidoptera: Lyonetiidae) on apple tree

Leaf-mining by insects usually have limited detrimental consequences on host-trees. Yet, Leucoptera malifoliella, a common leaf-mining moth on Rosaceae, is among those leaf-mining species that can exhibit occasional, sometimes even regular eruptive impacts on their hosts, in particular on apple trees. Here, we address the different causes involved in a particular case of nascent outbreak of this moth observed in the wild that is independently of the artificial conditions that prevail in densely planted apple orchards. Relevantly analyzing the reasons of eruptive outbreaks remains difficult however, as multiple causes are involved and their respective influences on mining-impact level are largely intermingled. Thus, in order to disentangle the respective contributions of each cause in quantitative terms, a specifically dedicated approach, the “Melba” procedure, was implemented and applied to the data issued from an exhaustive sampling of the foliage of the studied wild apple-tree. Out of the three main causal components of mining impact level, neither the average clutch-size nor the severity of mated females in selecting those leaves acceptable for oviposition substantially differ from the ranges of values encountered in other leaf-mining species having high but non eruptive mining impacts on their respective hosts. In fact, the recorded nascent outbreak is mainly attributable to the unusually high density of mated females attracted to the investigated host tree. Accordingly, metapopulation effects obviously play a major role in triggering the recorded eruptive impact.

Native birds exploit leaf-mining moth larvae using a new North American host, non-native Lonicera maackii

ABSTRACTI document observations of birds feeding on the larvae of the leaf-mining moth Phyllonorycter emberizaepenella (Gracillariidae) in Michigan, USA. The larvae were feeding on Amur honeysuckle, Lonicera maackii (Caprifoliaceae), a non-native plant that has not been documented as a host for any species of Gracillariidae in North America, and which has few records of use by invertebrate herbivores anywhere in its range. Most bird observations were of chickadees (Paridae), which have been previously recorded feeding on leaf-mining insects. Observations also include three species of warblers (Parulidae) and one kinglet (Regulidae), which represent the first documentation of these families feeding on leaf-mining moth larvae. I describe this novel multitrophic interaction between a non-native plant, a native insect, and native vertebrate predators, and discuss the potential spread of the moth in North America, its use as a food source by birds, and the ecological implications.

Dynamics and origin of cytokinins involved in plant manipulation by a leaf-mining insect.

Several herbivorous insects and plant-associated microorganisms control the phytohormonal balance, thus enabling them to successfully exploit the plant by inhibiting plant defenses and withdrawing plant resources for their own benefit. The leaf-mining moth Phyllonorycter blancardella modifies the cytokinin (CK) profile of mined leaf-tissues, and the insect symbiotic bacteria Wolbachia is involved in the plant manipulation to the benefit of the insect host. To gain a deeper understanding into the possible origin and dynamics of CKs, we conducted an extensive characterization of CKs in larvae and in infected apple leaves. Our results show the enhanced CK levels in mines, both on green and yellow leaves, allowing insects to control their nutritional supply under fluctuating environmental conditions. The spatial distribution of CKs within the mined leaves shows that hormone manipulation is strictly limited to the mine suggesting the absence of CK translocation from distant leaf areas toward the insect feeding site. Mass spectrometry analyses reveal that major CK types accumulating in mines and larvae are similar to what is observed for most gall-inducers, suggesting that strategies underlying the plant manipulation may be shared between herbivorous insects with distinct life histories. Results further show that CKs are detected in the highest levels in larvae, reinforcing our hypothesis that CKs accumulating in the mines originate from the insect itself. Presence of bacteria-specific methylthio-CKs is consistent with previous results suggesting that insect bacterial symbionts contribute to the observed phenotype. Our study provides key findings toward the understanding of molecular mechanisms underlying this intricate plant-insect-microbe interaction.

New species of leaf-mining Nepticulidae (Lepidoptera) from the Neotropical and Ando-Patagonian regions, with new data on host plants

The paper treats fifteen species of leaf-mining pygmy moths (Insecta, Lepidoptera, Nepticulidae) discovered in the Neotropics (British Virgin Islands, Belize, Costa Rica, Venezuela, and Ecuador), and Ando-Patagonian region (Argentina and Chile). Except for two species, all belong to Stigmella Schrank. Twelve species are new, and are named and described in the current paper: Stigmella apicibrunella Diškus & Stonis, sp. nov.; S. decora Diškus & Stonis, sp. nov.; S. unicaudata Remeikis & Stonis, sp. nov.; S. sanmartini Remeikis & Stonis, sp. nov.; S. patula Remeikis & Stonis, sp. nov.; S. torosa Remeikis & Stonis, sp. nov.; S. monstrata Remeikis & Stonis, sp. nov.; S. huahumi Remeikis & Stonis, sp. nov.; S. venezuelica Remeikis & Stonis, sp. nov.; S. virginica Remeikis & Stonis, sp. nov.; Fomoria miranda Diškus & Stonis, sp. nov.; and Hesperolyra robinsoni Stonis, sp. n. Newly discovered variation of male genitalia of the Andean Stigmella rudis Puplesis & Robinson, 2000 is briefly discussed, and the formerly poorly understood Stigmella hylomaga (Meyrick, 1931) is redescribed and documented with photographs for the first time. We also present more photographs and add some addtional information on Stigmella gallicola van Nieukerken & Nishida, a recently described gall-maker from Costa Rica.The paper also provides new host-plant data: some of the described (or redescribed) species are reported for the first time as leaf-miners on plants belonging to Euphorbiaceae (Acalypha padifolia Kunth), Salicaceae (Azara microphylla Hook. f.), Fabaceae (Inga spectabilis (Vahl) Willd. or I. edulis Mart.), Rhamnaceae (Colletia spinosissima J. F. Gmel.), Geraniaceae or Vivianiaceae (Rhynchotheca spinosa Ruiz & Pav.), and Asteraceae (Mutisia decurrens Cav.). All species treated in the paper are illustrated with photographs of the adults and genitalia, a distribution map, and also photographs of the leaf-mines and host plants when available.

A signature of tree health? Shifts in the microbiome and the ecological drivers of horse chestnut bleeding canker disease.

Host susceptibility to pathogens can be shaped by genetic, ecological, and evolutionary factors. The ability to predict the spread of disease therefore requires an integrated understanding of these factors, including effects of pests on pathogen growth and competition between pathogens and commensal microbiota for host resources. We examined interactions between the leaf-mining moth Cameraria ohridella, the bacterial causal agent of bleeding canker disease Pseudomonas syringae pv aesculi, and the bark-associated microbiota of horse chestnut (Aesculus hippocastanum) trees. Through surveys of >900 trees from 60 sites in the UK, we tested for ecological or life history predictors of leaf miner infestation, bleeding canker, or coinfection. Using culture-independent sequencing, we then compared the bark microbiomes from 46 trees to measure the association between microbiome composition and key ecological variables, including the severity of disease. Both pest and pathogen were found to respond to tree characteristics, but neither explained damage inflicted by the other. However, we found a clear loss of microbial diversity and associated shift in microbiome composition of trees as a function of disease. These results show a link between bark-associated microbiota and tree health that introduces the intriguing possibility that tree microbiota play key roles in the spread of disease.

Ti-catalyzed cross-cyclomagnesiation of 1,2-dienes in the stereoselective synthesis of insect pheromones

The Ti-catalyzed intermolecular cross-cyclomagnesiation reaction of aliphatic and oxygenated 1,2-dienes with Grignard reagents was used as the key step to develop facile stereoselective methods for preparation of the major components of citrus leafminer moth (Phyllocnistis citrella) pheromones as well as cotton pink bollworm (Pectinophora gossypiella) and fruit fly (Drosophila melanogaster) attractants.

Genetic variation in foundation species governs the dynamics of trophic interactions.

Various studies have demonstrated that the foundation species genetic diversity can have direct effects that extend beyond the individual or population level, affecting the dependent communities. Additionally, these effects may be indirectly extended to higher trophic levels throughout the entire community. Quercus castanea is an oak species with characteristics of foundation species beyond presenting a wide geographical distribution and being a dominant element of Mexican temperate forests. In this study, we analyzed the influence of population (He) and individual (HL) genetic diversity of Q. castanea on its canopy endophagous insect community and associated parasitoids. Specifically, we studied the composition, richness (S) and density of leaf-mining moths (Lepidoptera: Tischeridae, Citheraniidae), gall-forming wasps (Hymenoptera: Cynipidae), and canopy parasitoids of Q. castanea. We sampled 120 trees belonging to six populations (20/site) through the previously recognized gradient of genetic diversity. In total, 22 endophagous insect species belonging to three orders (Hymenoptera, Lepidoptera, and Diptera) and 20 parasitoid species belonging to 13 families were identified. In general, we observed that the individual genetic diversity of the host plant (HL) has a significant positive effect on the S and density of the canopy endophagous insect communities. In contrast, He has a significant negative effect on the S of endophagous insects. Additionally, indirect effects of HL were observed, affecting the S and density of parasitoid insects. Our results suggest that genetic variation in foundation species can be one of the most important factors governing the dynamics of tritrophic interactions that involve oaks, herbivores, and parasitoids.

Seasonal monitoring of the flight activity and the incidence of the groundnut leaf minerAproaerema modicella(Lepidoptera: Gelechiidae) at five sites in South Africa

The groundnut leaf miner, Aproaerema modicella (Deventer), invaded the African continent in 1998 and is devastating leaf miner pest of groundnut (Arachis hypogaea L.). There is limited information on this pest's ecology to facilitate the management strategies, and thus a study on its ecology was conducted at five sites in South Africa for two years. We monitored A. modicella flight activity using pheromone traps and larval infestation on various commercial (groundnut, soya bean (Glycine maxima L.), lucerne (Medicago sativa L.) and pigeon pea (Cajanus cajan L.)) and wild hosts in order to understand its dispersal and the role of alternative hosts in the carryover of A. modicella populations from one season to another. The data was correlated with site specific climatic data (rainfall, temperature and humidity). At all locations, groundnut leaf miner moths were caught in traps before crop planting. Though low in numbers, groundnut leaf miner moths were caught during winter at all locations other than Brits. Infestations on pigeon pea and lucerne were observed in March and April 2012. A significant negative association between temperature and A. modicella moth catches was observed at Nelspruit, while Vaalharts had a significant positive association between humidity and A. modicella moth catches. There was no significant correlation between any of the recorded environmental factors and moth catches at Manguzi and Brits. Monitoring A. modicella in the current study provided some useful ecological information regarding the pest which assisted in establishing the pest's peak and off-peak periods. This information should be useful in determining control strategies such as pesticide application schedules.

Novel Chemo-Attractants for Trapping Tomato Leafminer Moth (Lepidoptera: Gelechiidae).

The tomato leafminer moth, Tuta absoluta (Meyrick), is a devastating pest for tomatoes in Iran and throughout the world. This pest reduces tomato yields in farms and greenhouses. It appears some floral odors are significant attractants for tomato leafminer moth. In this study, the effects of three floral compounds, phenylacetaldehyde (PAA), acetic acid (AA), and 3-methyl-1-butanol (MB), were evaluated as trap attractants for tomato leafminer moth. The attractants were tested separately, combined, and blended in binary and tertiary. Lures were tested in delta and water pan traps under field conditions and compared with unbaited traps as controls. Results indicated that water pan traps caught more moths than delta traps. Also treatments with PAA + AA combined and the AA + MB blend were strongly attractive to tomato leafminer moth males compared with other treatments in this study.

Molecular Phylogeny, Revised Higher Classification, and Implications for Conservation of Endangered Hawaiian Leaf-Mining Moths (Lepidoptera: Gracillariidae: Philodoria)1

Abstract: The leaf-mining moth genus Philodoria Walsingham (Lepidoptera: Gracillariidae) is composed of 30 described species, all of which are endemic to the Hawaiian Islands. Philodoria is known to feed on 10 families of endemic Hawaiian host plants, with several species recorded only from threatened or endangered hosts. Beyond their dependence on these plants, little is known of their evolutionary history and conservation status. We constructed a molecular phylogeny of Philodoria to assess validity of its current subgeneric classification and to help guide future work on this threatened Hawaiian lineage. Mitochondrial and nuclear DNA sequences from three genes (CO1, CAD, EF-1α) combining for a total of 2,041 base pairs, were collected from 11 Philodoria species, incorporating taxa from both currently recognized subgenera. These data were analyzed using both parsimony and model-based phylogenetic approaches. Contrary to the most recent systematic treatment of Philodoria, our results indicate strongly tha...

Host plant preference and offspring performance of a leaf‐mining moth, Caloptilia fraxinella, on two Fraxinus species

AbstractThe preference‐performance or ‘mother‐knows‐best’ hypothesis states that female insects choose to oviposit on a host plant that increases the performance of their offspring. This positive link between host plant choice and larval performance is especially important for leaf miners with non‐motile larvae that are entirely dependent upon the oviposition choice of the female for host plant location. Preference and performance of the ash leaf coneroller, Caloptilia fraxinella (Ely) (Lepidoptera: Gracillariidae), a specialist on ash trees, Fraxinus spp. (Oleaceae), were tested in a series of laboratory and field experiments. Female C. fraxinella were exposed to two closely related hosts, black ash, Fraxinus nigra Marshall, and green ash, Fraxinus pennsylvanica Marshall var. subintegerrima (Vahl), in oviposition choice and wind tunnel flight experiments to determine which host is most attractive for oviposition. Caloptilia fraxinella females were inconsistent in host choice, yet performance of larvae was greater on green than black ash. In preference studies, C. fraxinella preferred to oviposit on black ash when leaflets were removed from the tree, but preferred intact green ash over black ash seedlings for oviposition and host location in a wind tunnel. In the field, however, more C. fraxinella visited black ash var. ‘Fallgold’ at leaf flush than green ash at the same sites. Age of the ash leaflet also influences oviposition in this leaf miner and females preferred new over old leaflets for oviposition. Performance of C. fraxinella larvae was evaluated in field and laboratory experiments and was greater on green ash than on black ash in both experiments based on larval survival and development time parameters. The stronger oviposition and host location preference in the field for black ash were not linked to enhanced performance of offspring, as green ash was the superior host, supporting higher larval survival and faster development. A stronger host location preference in the wind tunnel for green ash over black ash, however, suggests that under certain circumstances with this moth species, ‘mother (may) know best’.

Industrial pollution affects behaviour of the leafmining moth Stigmella lapponica

AbstractWe explored the impacts of industrial air pollution on the behaviour of the leafmining moth Stigmella lapponica (Wocke) (Lepidoptera: Nepticulidae) by comparing the characteristics of larval gallery mines in mountain birch [Betula pubescens ssp. czerepanovii (Orlova) Hämet‐Ahti (Betulaceae)] leaves collected from unpolluted forests and from heavily polluted industrial barrens surrounding the copper‐nickel smelter in Monchegorsk in north‐western Russia. Population density of S. lapponica, survival of larvae, length of the completed mine, and width of its terminal part did not differ between polluted and unpolluted sites. Females in unpolluted sites only rarely (16%) oviposited on the apical half of the leaf and the larval mortality in mines that started in the apical part of the leaf was 83%. A significantly larger (38%) proportion of mines started in the apical half of the leaves in polluted sites, and the larval mortality in these mines was only 45%. The between‐habitat difference in the choice of the oviposition sites by S. lapponica is the first demonstration of the adaptive plasticity of oviposition behaviour in a leafmining insect. This difference was not explained by specific leaf weight which did not vary within leaves. Larvae mining in polluted leaves extended 25% farther between turns, and the galleries turned more sharply than in unpolluted leaves. This result confirms the abilities of leafmining larvae to evaluate the quality of the ingested food and adjust their behaviour accordingly. Thus, pollution modifies both the preference of S. lapponica females for oviposition sites within a birch leaf and the behaviour of S. lapponica larvae mining these leaves. This is one of the first records of changes in insect behaviour in natural environment disturbed by industrial pollution.

The Relationship between Polarized Moonlight and the Number of Pest Microlepidoptera Specimens Caught in Pheromone Traps

AbstractPheromone traps were deployed in Borsod-Abaúj-Zemplén County (Hungary) between 1982 and 1988, in 1990 and also between 1993 and 2013. These traps attracted 8 Microlepidoptera species:Phyllonorycter blancardella, P. corylifoliella,Anarsia lineatella,Eupoecilia ambiguella,Lobesia botrana,Grapholita funebrana,G. molestaandCydia pomonella. We examined the trapping data of these species in the context of lunar phases and polarized moonlight. Catches of the European Vine Moth (Lobesia botrana) and the Codling Moth (Cydia pomonella) were higher in the First Quarter, whereas catches of Peach Twig Borer (Anarsia lineatella), Vine Moth (Eupoecilia ambiguella), Plum Fruit Moth (Grapholita funebrana) and Oriental Fruit Moth (Grapholita molesta) were larger in the Last Quarter. Catches of the other two species, the Spotted Tentiform Leafminer (Phyllonorycter blancardella) and Hawthorn Red Midget Moth (P. corylifoliella), were higher in both the First and Last Quarters. When using pheromone traps, insects do not fly to a light source, so moonlight does not modify either the catching distance or flight activity. However, at high levels of polarized moonlight, pheromone trap catches will increase, as in the case of light-trap catches. The results are comprehensible when one considers that the target species can fly both during the daytime and also at night.

Correlation between the green-island phenotype and Wolbachia infections during the evolutionary diversification of Gracillariidae leaf-mining moths.

Internally feeding herbivorous insects such as leaf miners have developed the ability to manipulate the physiology of their host plants in a way to best meet their metabolic needs and compensate for variation in food nutritional composition. For instance, some leaf miners can induce green-islands on yellow leaves in autumn, which are characterized by photosynthetically active green patches in otherwise senescing leaves. It has been shown that endosymbionts, and most likely bacteria of the genus Wolbachia, play an important role in green-island induction in the apple leaf-mining moth Phyllonorycter blancardella. However, it is currently not known how widespread is this moth-Wolbachia-plant interaction. Here, we studied the co-occurrence between Wolbachia and the green-island phenotype in 133 moth specimens belonging to 74 species of Lepidoptera including 60 Gracillariidae leaf miners. Using a combination of molecular phylogenies and ecological data (occurrence of green-islands), we show that the acquisitions of the green-island phenotype and Wolbachia infections have been associated through the evolutionary diversification of Gracillariidae. We also found intraspecific variability in both green-island formation and Wolbachia infection, with some species being able to form green-islands without being infected by Wolbachia. In addition, Wolbachia variants belonging to both A and B supergroups were found to be associated with green-island phenotype suggesting several independent origins of green-island induction. This study opens new prospects and raises new questions about the ecology and evolution of the tripartite association between Wolbachia, leaf miners, and their host plants.

A morphological reappraisal of the immature stages and life history of Elachista synethes Meyrick (Lepidoptera, Elachistidae), an Australian leaf miner alien to Chile

Elachista synethes was recently recognized as an alien species in northern Chile, where its larvae mine the rescue grass Bromus catharticus (Poaceae). In order to provide the necessary information to allow field detection of E. synethes during early ontogeny, we conducted a morphological reappraisal of the immature stages of this leaf-miner moth, based on light and scanning electron microscopy, including the first descriptions of the egg and the first-instar larva. This is the first report of the existence of an apodal early larva for a species of Elachista Treitschke. The legs and prolegs are absent in the first two instars, but are well developed in the last two. Additional observations on the life history are also provided, including a description of the mine.