Laboratory life history and field observations of Poliopaschia lithochlora (Lower) (Lepidoptera: Pyralidae), a potential biological control agent for Melaleuca quinquenervia (Myrtaceae)

Abstract  Melaleuca quinquenervia (Cav.) S.T. Blake, Australian broad-leaved paperbark, has become a serious weed in southern Florida. Poliopaschia lithochlora (Lower) is a promising candidate as a potential biological control agent, and this study describes laboratory and field observations of the life history of this moth. Eggs are laid in small batches, mainly on the surface of leaves, and larvae are voracious leaf feeders, concealed in tubes that are usually found in small colonies attached to leaves and stems. Larvae move from these tubes to feed on surrounding leaves, and saplings and suckers are frequently defoliated. Prepupae form sealed bulbs in the larval tubes in which they pupate. Adult females are mainly active and oviposit at night. Development from egg to adult occurs in approximately 80 d. Field populations appear to be regulated by several egg and larval parasites. Because this moth severely damages saplings and suckers of M .  quinquenervia , prefers low-lying humid sites, and can be successfully mass reared, it is rated highly for its potential as a biological control agent.     

Host Plant Quality Factors That Influence the Growth and Development of Oxyops vitiosa, a Biological Control Agent of Melaleuca quinquenervia

The Australian weevil Oxyops vitiosa was released in 1997 in Florida as a biological control agent of Melaleuca quinquenervia. The larvae of this agent are flush-feeders, found only on the growing tips of their host. Knowledge of this restriction to feeding on the growing tips and other nutritional requirements may assist in the establishment and dispersal of this species. Therefore, O. vitiosa survival was assessed when neonates were fed M. quinquenervia leaves from branches that had dormant buds or emerging bud leaves. Additionally, the influence of leaf quality from different sites and within sites was determined by the feeding of neonates emerging bud leaves collected at three sites and from three leaf qualities (poor, intermediate, and high). Within-site leaf qualities were described in the field by leaf color and in the laboratory by percentage dry mass and nitrogen. Larval survival was lowest when fed leaves from branches that had dormant buds. Associated with this low survival were high leaf toughness and percentage dry mass. When larvae were fed emerging bud leaves, most of the variation in larval survival and performance was attributed to differences in within-site plant quality. Generally, the highest-quality leaves had relatively low percentage dry mass and high percentage nitrogen. Larval survival generally decreased when fed the poor-quality leaves, and in one site, the intermediate-quality leaves. Larvae required less time to develop to adults when fed the high-quality leaves. Development time increased in females but not in males when the larvae were fed the poor-quality leaves. Adult biomass of both females and males generally increased when the larvae were fed the high-quality leaves from two of the three sites. The results indicate that the larvae of O. vitiosa are restricted to feeding on flush foliage with low toughness. Additionally, variations in foliar percentage dry mass and nitrogen influence larval survival and performance. This knowledge benefited the development of mass-production nursery sites and the selection of suitable release sites, which facilitated the establishment of this biological control agent.     

Chemical detoxification vs mechanical removal of host plant toxins in Eucalyptus feeding sawfly larvae (Hymenoptera: Pergidae)

The essential oils that characterize the eucalypts and related Myrtaceae pose a challenge for herbivores. Phytophagous insects that feed on oil-rich Myrtaceae have developed specific mechanisms to deal with these oils, some of which are notoriously toxic (e.g. 1,8-cineole). Some of the eight Australian subfamilies in the sawfly family Pergidae are associated exclusively with Eucalyptus and Melaleuca species that often have high concentrations of essential oils. Unexpectedly, the Perginae and Pterygophorinae use different mechanisms to deal with the same toxic components in their respective host plants. Larvae of the Perginae have the inner surface of their mandibles equipped with soft brush-like structures that are unique among phytophagous insects in general. The proposed role of these ancillary mandibular structures in separating leaf oils from nutritive plant matter could be confirmed in experiments with larvae of two pergine species. The oil sequestration is, however, incomplete and chemical gut content analyses by gas-chromatography (GC) revealed that 1,8-cineole does enter the midgut and is metabolised to hydroxycineole. Although the related Pterygophorinae also feed mainly on oil-rich Myrtaceae, they do not sequester the oil and lack morphological structures on their mandibles. Chemical analysis of the gut content of two pterygophorine species showed that they rely solely on chemical detoxification of the relevant plant compounds, with GC demonstrating that the 1,8-cineole is removed far more rapidly and completely than in the pergine species.     

Quarantine host range studies with Lophyrotoma zonalis, an Australian sawfly of interest for biological control of melaleuca, Melaleuca quinquenervia, in Florida, U.S.A.

The Australian melaleuca tree, Melaleuca quinquenervia (Cav.) S. T. Blake (Myrtaceae), has naturalized in southern Florida,U.S.A., and is now one of that regions most important weeds.Primarily a weed of wetlands, it also infests neighboring drierareas. Current efforts to restore the South Florida ecosystem arethreatened by the continuing range expansion of melaleuca andother weeds. In an effort to supplement the current chemical andcultural control methods for melaleuca, a search for potentialbiological control agents was begun in Australia in 1986. Thesawfly, Lophyrotoma zonalis, was determined after extensive fieldand laboratory studies to have potential as a biological controlagent. Larvae of L. zonalis eat leaves and occasionally defoliatelarge trees in Australia. Host range studies were conducted in aFlorida quarantine facility with native and cultivated plantspecies. Multi-choice and no-choice oviposition tests wereconducted with 36 species in the Myrtaceae and with 18 species inother families. Larvae developed to prepupae and adults from theeggs oviposited on 23 species of Myrtaceae only on 3 species ofbottlebrushes, Callistemon. Medium-sized larvae were tested forfeeding on bouquets of plant cuttings and on potted plants. Theyare the stage that might wander from defoliated trees. Noticeablefeeding, but much less than on melaleuca, was restricted to theMyrtaceae, except for a few individual larvae that fed on waxmyrtle, Myrica cerifera. Medium-sized larvae became prepupae onlyon Melaleuca decora (73%) and on wax myrtle (10%). However,neither species received eggs in the oviposition tests. Thesestudies confirmed the narrow host range of L. zonalis aspreviously reported from field and laboratory studies inAustralia.     

Reticulate evolution in the natural range of the invasive wetland tree species Melaleuca quinquenervia

The Melaleuca leucadendra complex (broad-leaf paperbarks; Myrtaceae) is a dominant component of the tropical and sub-tropical biota of Australia, particularly in wetlands of high conservation significance. In Florida and other parts of the Americas, however, one member of the group (Melaleuca quinquenervia) is a serious ecological and economic weed. Understanding the relationships and evolution of the group is integral to both conservation and biocontrol efforts. Although the complex is currently considered to include up to 14 species, there has been some concern over taxonomic boundaries within the complex because most species are circumscribed only by combinations of characters, each of which also occurs in other species. Here, DNA sequence data derived from the chloroplast and two nuclear regions are used to explore the relationships of M. quinquenervia. We find little evidence for clear species boundaries within the M. leucadendra complex in general, with regional sharing of chloroplast haplotypes across morphologically defined taxa, indicating asymmetrical introgression or retention of ancestral haplotypes (lineage sorting). Phylogenies were further confounded by the recovery of multiple copies of both nuclear regions sequenced (ITS and rpb2) from many individuals. There was no clear evidence of polyploidy or pseudogenes, but multiple duplications of rpb2 could not be ruled out. Parsimony networks of the nuclear ITS region show some clustering of haplotypes by morphospecies but there is also evidence of both hybridisation and recombination. Signals of introgression were also evident in rpb2, supporting an hypothesis of recent or ongoing gene flow between M. quinquenervia and other members of the M. leucadendra complex. Both relaxed and fixed molecular-clock dating estimate the introgression to have occurred sometime within the past seven million years (95% CI: 0.7-18). The New Caledonian population of M. quinquenervia appears to have been established by dispersal from Australia during this period. M. quinquenervia is found to have alleles closely related to multiple different morphotaxa within the M. leucadendra complex, suggesting considerable past introgression into this taxon from some other members of the M. leucadendra complex, and this has implications for biocontrol efforts. The M. leucadendra complex appears to reflect early to intermediate stages of speciation, possibly driven by different ecologies.     

Modification of hormonal balance in larvae of the corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae) due to sublethal Bacillus thuringiensis protein ingestion

Bacillus thuringiensis (Bt) corn, Zea mays L., is highly efficient against the corn borer Sesamia nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae) when the larvae feed only on the transgenic plants. However, when they feed on Bt leaves during only part of their development, thus ingesting sublethal amounts of Bt toxins, some larvae survive. A previous study reported a prolonged development and precocious diapause induction in larvae fed on a diet with sublethal amounts of Cry1Ab protein. To determine whether these effects were accompanied by a modification of the hormonal balance, S. nonagrioides larvae were fed on sublethal amounts of Bt protein provided in Bt leaves or in the diet. The larvae that survived had higher levels of juvenile hormone (JH), whereas their level of ecdysteroids did not increase sufficiently to allow pupation, leading to a longer larval development and more larval molts. This response may be considered a defense mechanism that allows some larvae to survive toxin ingestion; it is similar the response to insecticidal toxins or viruses observed in other larvae. Changes in the hormone levels in diapausing larvae were undetectable, probably because these changes were masked by the higher level of JH in the hemolymph of diapausing larvae and because of lack of ecdysteroid titer increase, a phenomenon that is usually observed a few days before pupation in nondiapausing larvae. These results should be taken into account in the establishment of non-Bt refuges to prevent development of Bt-resistance in S. non-agrioides populations.     

Effects of class-specific,synthetic, and natural proteinase inhibitors on life-history traits of the cotton bollworm Helicoverpa armigera

Herbivorous insects have more difficulty obtaining proteins from their food than do predators and parasites. The scarcity of proteins in their diet requires herbivores to feed voraciously, thus heavily damaging their host plants. Plants respond to herbivory by producing defense compounds, which reduce insect growth, retard development, and increase mortality. Herbivores use both pre- and postdigestive response mechanisms to detect and avoid plant defense compounds. Proteinase inhibitors (PIs) are one example of plant compounds produced as a direct defense against herbivory. Many insects can adapt to PIs when these are incorporated into artificial diets. However, little is known about the effect of PIs on diet choice and feeding behavior. We monitored the diet choice, life-history traits, and gut proteinase activity of Helicoverpa armigera larvae using diets supplemented with synthetic and natural PIs. In choice experiments, both neonates and fourth-instar larvae preferred the control diet over PI-supplemented diets, to varying degrees. Larvae that fed on PI-supplemented diets weighed less than those that fed on the control diet and produced smaller pupae. Trypsin-specific PIs had a stronger effect on mean larval weight than did other PIs. A reduction of trypsin activity but not of chymotrypsin activity was observed in larvae fed on PI-supplemented diets. Therefore, behavioral avoidance of feeding on plant parts high in PIs could be an adaptation to minimize the impact of this plant's defensive strategy.     

Effects of diet-deprivation and physical stimulation on the feeding behaviour of the larvae of the silkworm, Bombyx mori

Continuous observations of larvae of the silkworm, Bombyx mori, revealed that feeding occurred at regular intervals throughout larval development. To investigate possible factors influencing meal-timing, the behaviours of diet-deprived Bombyx larvae were also analysed. Diet-deprivation resulted in longer durations of the first meals after diet replacement, but did not affect feeding patterns. Furthermore, long-term diet-deprivation promoted wandering behaviour and a consequent delay in feeding after diet replacement. Under diet-deprivation conditions, meal-starts appeared to be inducible by defecation and physical stimulation. However, stimulation-induced meal-starts were dependent on the time elapsed since the larvae's previous meals. Provided that more than 1h had elapsed since their previous meals, larvae could be induced to feed by defecation and tapping. At less than 1h post-meal, larvae were less likely to begin feeding after defecation or physical stimulation. Activated locomotions such as wandering and feeding were observed in the long-term diet-deprived larvae only after diet blocks were replaced, while long-term diet-deprived larvae did not show activated locomotion during the absence of diet blocks. Collectively, these data suggest that a combination of elevated locomotion activity and the presence of diet may be necessary for the initiation of feeding in diet-deprived larvae.     

Ecology and management of Melaleuca quinquenervia,an invader of wetlands in Florida,U.S.A.

Invasive weeds are potent agents of environmental change. Wetlands are valuable environments that frequently are impinged by a variety of threats including invasive weeds. Melaleuca quinquenervia (Cav.) S.T. Blake (broad-leaved paperbark), though experiencing major diminishment of native populations in Australia, is naturalized and highly invasive in most wetland habitats of south Florida, U.S.A., where it infests more than 202,000 ha. Wetlands in south Florida, including such renowned areas as the Everglades, are being transformed into M. quinquenervia swamps, with major environmental and economic impacts. Current management methods include herbicides, mechanical or hand removal of plants, flooding, and prescribed burning. Insufficient information, high costs, non-target impacts, and the resilience of M. quinquenervia (trunk and root sprouts and massive canopy seed banks) greatly constrain the effectiveness of these control methods. Biological control offers long-term management potential, most likely by reducing the rate of spread and the vitality and growth rate of plants, thus rendering them more vulnerable to other environmental stresses and control methods. The leaf weevil Oxyops vitiosa Pascoe, a natural enemy of M. quinquenervia in Australia, will likely be the first biocontrol agent released against the weed in Florida. More information is needed, especially ecological data, to better understand the invasiveness of M. quinquenervia in Florida and to facilitate its management there.     

Use of the characters of trophic specialization in the taxonomy of the broad-winged moths (Lepidoptera: Oecophoridae,Chimabachidae, Amphisbatidae,Depressariidae)

Trophic specialization of the larvae of Oecophorid moths from the Palaearctic Region is investigated. The former family Oecophoridae (sensu lato) is presently considered a polyphyletic group and is divided into the separate families Oecophoridae sensu stricto (with subfamilies Deuterogoniinae, Oecophorinae and Pleurotinae), Chimabachidae, Amphisbatidae and Depressariidae. Xyloryctidae, Stathmopodidae, Ethmiidae and Autostichidae are also considered as separate families. In these taxa, two main modes of feeding occur: on dead plants and on living plants. The first mode is inhered in Oecophoridae (in subfamily Pleurotinae both the modes are known) and Autostichidae. The family Amphisbatidae occupies an intermediate position as the larvae of Amphisbatini use dead leaves, whereas the larvae of Fuchsiini and Hypercalliini use green plants. The rest of the families associate with living plants. Trophic specialization in the families formerly included in Oecophoridae mostly corresponds to the taxonomic division based mainly on the morphological characters, although some exceptions are known, e. g. feeding of larvae of Carcina quercana (Oecophoridae, Oecophorinae, Carcinini) on living leaves.     

Egg and larval load assessment and its influence on oviposition behaviour of the leaf beetle Galerucella nymphaeae

The oviposition behaviour of the water-lily beetle Galerucella nymphaeae was examined. This species is a specialist herbivore on the floating leaves of nymphaeids Nymphaeaceae and especially on the yellow water-lily, Nuphar lutea. Females lay their eggs in clutches on the leaves, and after hatching, the larvae feed on the leaves. The quality of the leaves decreases quickly after the larvae hatch, and eventually the leaves will sink below the water surface, whereupon the eggs, 1st-instar larvae and pupae are killed by drowning. The influence of conspecific eggs, larvae and feeding tracks on the oviposition preferences of the beetles was tested. Females were allowed to choose between fresh leaves and leaves with conspecific eggs and larvae as well as between leaves with larvae and leaves with feeding tracks but no larvae. An attempt was also made to determine whether eggs and larvae affect the oviposition rate of females when they are not given the opportunity to oviposit on untouched leaves. The results indicate that females tended to avoid leaves with conspecific larvae or to exhibit a decreased oviposition rate on such leaves. Females also avoided conspecific eggs, although the oviposition rate was not influenced by the presence of conspecific eggs. When females were allowed to choose between leaves with larvae and leaves with feeding tracks, possible discrimination against leaves with larvae just fails to reach the 5% level.     

Morphological and anatomical changes of Melaleuca cajuputi under submergence

Melaleuca cajuputi is a woody plant of the Myrtaceae which is a dominant species in tropical peat swamps in southern Thailand, where the groundwater level fluctuates greatly. Although the current year seedlings are likely submerged, their adaptive responses have never been studied. The objective of the present study was to examine their responses to submergence, and especially their morphological and anatomical changes. Not only did the seedlings of M. cajuputi survive submergence for 56 days, but they could also increase their dry weight, shoot length, and leaf number during submergence. These growth responses to submergence indicate that the seedlings of M. cajuputi could make photosynthetic production under water. The leaves that developed under water were heterophyllous “aquatic leaves” that appear to represent adaptations to improve the uptake of gases from the water. Intercellular spaces in the stems and leaves were more strongly developed in the submerged seedlings than in non-submerged seedlings with the shoot and leaves in the air. The intercellular spaces appear to be schizogenous aerenchyma that facilitates gas exchange. The growth responses and anatomical responses in stems and leaves to submergence, which were found in M. cajuputi, are commonly known in herbaceous plants with amphibious characteristics, but had not been reported in woody plants. And our results suggest that M. cajuputi adapts to submergence similarly to other amphibious plants, thereby ensuring continuing biomass production.     

Seasonality and genetic architecture of development time and body size of the birch feeding sawfly Priophorus pallipes

We tested, using the sawfly Priophorus pallipes feeding on leaves of mountain birch, whether the expression of genetic (co)variation of larval development time and body size can be altered by exposing larvae to diets with differential seasonal changes in quality. In nature, larvae feed mainly on mature leaves, but occasionally they are forced to consume senescing leaves. Sixty families were assayed on three experimentally simulated diets: mature leaves of high quality, senescing leaves of rapidly declining quality, and senesced leaves of low quality. The intuitively obvious positive phenotypic and genetic correlations between development time and final mass were observed when the larvae consumed leaves of stable high quality, but low and declining food quality prevented long-growing individuals and families from achieving high final mass, switching the correlations to close to zero or negative in these treatments. The amount of genetic variation for body size showed a non-linear change across the diet quality gradient, whereas genetic variation for development time increased with decreasing diet quality. The among-trait difference in the degree reaction norms crossed along the diet gradient caused the changes in the expression of genetic (co)variation within the environments. Our results show that seasonally varying diet quality induces dramatic changes in the genetic (co)variation of development time and body size, and that simultaneous analysis of reaction norms and environment-specific expression of genetic (co)variation is necessary for the understanding of the genetic characteristics underlying the construction of phenotypes in heterogeneous environments.     

Cross habituation to feeding deterrents and acceptance of a marginal host plant byPieris rapae larvae

Sensitivity of the cabbage butterfly,Pieris rapae L. to feeding deterrents was compared for larvae reared on different food sources under laboratory conditions. Since cabbage-reared larvae normally reject nasturtium,Tropaeolum majus L., the effects of previous exposure to allelochemicals on larval acceptance or rejection of this plant were also examined. When compared with cabbage-reared larvae, nasturtium-reared larvae were less sensitive to feeding deterrents including cymarin, erysimoside and 2-O-β-d-glucosyl cucurbitacin E. Nasturtium-reared larvae were insensitive to chlorogenic acid, which was deterrent to cabbage-reared larvae. Feeding by larvae reared on a wheat germ diet was not deterred by these compounds. The results indicate that dietary experience can extensively affect larval sensitivity to feeding deterrents and that cross habituation of larvae to deterrents occurs in response to certain chemical constituents of nasturtium and wheat germ diet. Digitoxin, however, proved to be an exception. Larvae reared on either nasturtium or wheat germ diet were as sensitive to digitoxin as those reared on cabbage. Previous results have shown that rejectionof nasturtium by cabbage-reared larvae is due to the presence of strong feeding deterrents in this plant. However, more than 50% of 2nd instar larvae reared from neonate on cabbage leaves treated with strophanthidin, cymarin, erysimoside, digitoxigenin and digitoxin accepted nasturtium as a food source. 2-O-β-d-glucosyl cucurbitacin E, 2-O-β-d-glucosyl cucurbitacin I and rutin were also active in causing larvae to feed on nasturtium. Thus dietary exposure to unrelated plant chemicals can profoundly affect insect acceptance of a plant that contains feeding deterrents.     

Intraguild predation among plant pests: western flower thrips larvae feed on whitefly crawlers

Omnivores obtain resources from more than one trophic level, and choose their food based on quantity and quality of these resources. For example, omnivores may switch to feeding on plants when prey are scarce. Larvae of the western flower thrips Frankiniella occidentalis Pergande (Thysanoptera: Thripidae) are an example of omnivores that become predatory when the quality of their host plant is low. Western flower thrips larvae usually feed on leaf tissue and on plant pollen, but may also attack eggs of predatory mites, their natural enemies, and eggs of the two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae), one of their competitors. Here, we present evidence that western flower thrips larvae prey on Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae), another competitor for plant tissue. We tested this on two host plant species, cucumber (Cucumis sativa L.), considered a host plant of high quality for western flower thrips, and sweet pepper (Capsicum annuum L.), a relatively poor quality host. We found that western flower thrips killed and fed especially on whitefly crawlers and that the incidence of feeding did not depend on host-plant species. The developmental rate and oviposition rate of western flower thrips was higher on a diet of cucumber leaves with whitefly crawlers than on cucumber leaves without whitefly crawlers, suggesting that thrips do not just kill whiteflies to reduce competition, but utilize whitefly crawlers as food.     

Antioxidant enzymes of the black swallowtail butterfly,Papilio polyxenes,and their response to the prooxidant allelochemical,quercetin

The black swallowtail butterfly larvae, Papilio polyxenes, are specialist feeders that have adapted to feeding on plants containing high levels of prooxidant allelochemicals. Third, fourth, and fifth instar larvae were tested for their antioxidant enzyme activities, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPOX), using 850-g supernatants from whole-body homogenates. The overall antioxidant enzyme profile for P. polyxenes was high compared to other insects, with activities ranging as follows: SOD, 1.1–7.5; CAT, 124–343; GR, 1.0–7.5; and GPOX, 0 units. To determine whether these antioxidant enzymes were inducible, P. poly xenes larvae were given a prooxidant challenge by dipping parsley leaves (their diet in the initial studies) in solutions of quercetin, such that the leaves became coated with this prooxidant flavonoid. Mid-fifth instar larvae fed on quercetin-coated leaves were assayed for antioxidant enzyme activities as was previously done with the larvae fed the standard diet. Food consumption and quercetin intake were monitored. SOD activity was increased almost twofold at the highest quercetin concentration tested. CAT and GR activity, on the other hand, were inhibited by increased quercetin consumption, with GR activity completely inhibited at the highest quercetin concentration after 12 h of feeding. GPOX activity, not present in control insects, was also not inducible by a quercetin challenge. These studies point out the key role that the antioxidant enzymes play in insect defenses against plant prooxidants.     

Feeding damage by larvae of the mustard leaf beetle deters conspecific females from oviposition and feeding

Herbivorous insects may be informed about the presence of competitors on the same host plant by a variety of cues. These cues can derive from either the competitor itself or the damaged plant. In the mustard leaf beetle Phaedon cochleariae (Coleoptera, Chrysomelidae), adults are known to be deterred from feeding and oviposition by the exocrine glandular secretion of conspecific co-occurring larvae. We hypothesised that the exocrine larval secretion released by feeding larvae may adsorb to the surface of Chinese cabbage leaves, and thus, convey the information about their former or actual presence. Further experiments tested the influence of leaves damaged by conspecific larvae, mechanically damaged leaves, larval frass and regurgitant on the oviposition and feeding behaviour of P. cochleariae. Finally, the effect of previous conspecific herbivory on larval development and larval host selection was assessed. Our results show that (epi)chrysomelidial, the major component of the exocrine secretion from P. cochleariae larvae, was detectable by GC-MS in surface extracts from leaves upon which larvae had fed. However, leaves exposed to volatiles of the larval secretion were not avoided by female P. cochleariae for feeding or oviposition. Thus, we conclude that secretion volatiles did not adsorb in sufficient amounts on the leaf surface to display deterrent activity towards adults. By contrast, gravid females avoided to feed and lay their eggs on leaves damaged by second-instar larvae for three days when compared to undamaged leaves. Mechanical damage of leaves and treatment of artificially damaged leaves with larval frass or regurgitant did not affect oviposition and feeding of P. cochleariae. Since no adverse effects of previous herbivory on larval development were detected, we suggest that female P. cochleariae avoid Chinese cabbage leaves damaged by feeding larvae for other reasons than escape from competition or avoidance of direct negative effects that result from consuming induced plant material.