Chew and spit: tree-feeding notodontid caterpillars anoint girdles with saliva

Caterpillars of the notodontid Oedemasia leptinoides (formerly Schizura) use their mandibles to cut shallow girdles that encircle the petioles and stems of tree hosts. When girdles are complete, the larvae bathe the girdle surface with fluid. We test whether the fluid originates from the labial salivary glands or ventral eversible gland by blocking the openings to the glands and observing whether fluid is still released onto the girdles. Only larvae with functional labial salivary glands anointed girdles with fluid. Analysis of girdle rinses for a prominent salivary enzyme, glucose oxidase, confirmed that larvae apply saliva and documented that application occurs primarily at the end of girdling. We propose that girdling by notodontids, together with related furrowing and leaf-clipping behaviors exhibited by diverse caterpillar groups, serve at least in part to introduce salivary components to exposed vascular tissues; these compounds presumably function to suppress plant defensive responses normally elicited by caterpillar feeding.     

Theroa zethus Caterpillars Use Acid Secretion of Anti-Predator Gland to Deactivate Plant Defense

In North America, notodontid caterpillars feed almost exclusively on hardwood trees. One notable exception, Theroa zethus feeds instead on herbaceous plants in the Euphorbiaceae protected by laticifers. These elongate canals follow leaf veins and contain latex under pressure; rupture causes the immediate release of sticky poisonous exudate. T. zethus larvae deactivate the latex defense of poinsettia and other euphorbs by applying acid from their ventral eversible gland, thereby creating furrows in the veins. The acid secretion softens the veins allowing larvae to compress even large veins with their mandibles and to disrupt laticifers internally often without contacting latex. Acid secretion collected from caterpillars and applied to the vein surface sufficed to create a furrow and to reduce latex exudation distal to the furrow where T. zethus larvae invariably feed. Larvae with their ventral eversible gland blocked were unable to create furrows and suffered reduced growth on poinsettia. The ventral eversible gland in T. zethus and other notodontids ordinarily serves to deter predators; when threatened, larvae spray acid from the gland orifice located between the mouthparts and first pair of legs. To my knowledge, T. zethus is the first caterpillar found to use an antipredator gland for disabling plant defenses. The novel combination of acid application and vein constriction allows T. zethus to exploit its unusual latex-bearing hosts.     

Biological Control of the Pecan Weevil,Curculio caryae (Coleoptera: Curculionidae), with Entomopathogenic Nematodes

Steinernema carpocapsae (Weiser) strain A11, S. feltiae (Filipjev) strain SN, and Heterorhabditis bacteriophora Poinar strains HP88 and Georgia were tested for their efficacy as biological control agents of the pecan weevil, Curculio caryae (Horn), in pecan orchard soil-profile containers under greenhouse conditions. Percentage C. caryae parasitism by S. carpocapsae and H. bacteriophora strain HP88 and Georgia was consistently poor when applied either prior to or following C. caryae entry into the soil, suggesting that these nematode species and (or) their enterobacteria are poor biological control agents of weevil larvae. Soil taken 21 days following application of S. carpocapsae or H. bacteriophora strain HP88 induced a low rate of infection of Galleria mellonella larvae, whereas soil that had been similarily treated with H. bacteriophora strain Georgia induced a moderate rate of infection. Percentage C. caryae parasitism by S. feltiae was consistently low when applied following C. caryae entry into the soil and was inconsistent when applied as a barrier prior to entry of weevil larvae into the soil. Soil taken 21 days following application of S. feltiae induced a high rate of infection of G. mellonella larvae.     

Dynamic changes in petiole specific conductivity in red maple (Acer rubrum L.), tulip tree (Liriodendron tulipifera L.) and northern fox grape (Vitis labrusca L.)

Diurnal variation in petiole specific hydraulic conductivity and simultaneous measurements of leaf water potential were recorded in red maple, tulip tree and fox grape. Petiole specific conductivity was determined from in situ measurements of water flow into the distal (leaf‐bearing) end of an attached petiole as a function of applied hydrostatic pressure and petiole dimensions. The hydraulic properties of the petiole dominated the measurements, indicating that this technique can be used for rapid estimates of petiole hydraulic conductivity. There was a significant decrease in petiole specific conductivity associated with increasingly more negative leaf water potentials in maple and tulip tree, but not in grape. Petiole specific conductivity increased during the afternoon while the plant was actively transpiring and the xylem sap was under tension. The recovery of petiole conductivity during the afternoon suggests that hydraulic conductivity reflects a dynamic balance between a loss of hydraulic conductivity with increasing water stress, and its restoration as tension within the xylem decreases. Three experimental manipulations were applied to red maple and tulip tree to examine the sensitivity of diurnal changes in petiole conductivity to various physiological perturbations. Both phloem girdling and application of HgCl₂ to the transpiration stream resulted in a marked decrease in the degree to which petiole specific conductivity recovered as xylem tension relaxed during the afternoon. Delivery of a surfactant to the xylem, however, did not significantly alter the relation between leaf water potential and petiole hydraulic conductivity.     

The gut bacteria symbionts from the monophagous insect Acrobasis nuxvorella produce tannase for the digestion of Carya illinoinensis tannins

Acrobasis nuxvorella Neuzing (Lepidoptera: Pyralidae), or the Pecan Nut Casebearer (PNC), is a monophagous pecan nut [Carya illinoinensis (Wang) K. Koch] herbivore. This insect has caused major crop losses, despite pecan nut trees producing a high concentration of tannins, which are deterrent compounds for insects. The PNC consumes and processes tannin-rich tissues without any negative effects on its nutrition. Certain bacterial species of the herbivore gut microbiota have been proven to produce tannase, therefore, we hypothesize that the PNC has symbiotic bacteria that produce tannase for the digestion of tannin contained in their food.In this work, live PNC larvae were extracted from pecan nutlets. The larval guts were dissected and their contents were cultured to obtain the cultivable bacteria. A total of 224 bacterial isolates were recovered, 19 of which tested positive for tannase activity. Isolates LB66, LB24, TT29, and TP8 displayed comparable activity to the control strain Pseudomonas aeruginosa. Further enzyme semi-purification steps showed specific tannase activity values in the range of 39.5–3.7 U/mg of protein. The isolates were identified as Bacillus pumilus strain TT29, Bacillus pumilus strain LB66, Bacillus pumilus strain LB24 and Sthaphylococcus warneri (TP8) by 16S ribosomal RNA gene sequencing. Our findings indicate that PNC larvae contain gut bacteria able to produce tannase that hydrolyze the galloyl ester bonds in tannins.     

Birds girdling activity on exotic tree species as a form of adaptive behavior?

Four tree species in the Kostelec n. ?. l. arboretum (Czech Republic) have been repeatedly damaged by Dendrocopos medius. The unique aspect of this otherwise common behavior called girdling consists in regularly visiting the same trees every spring, although there are more than 1.200 tree species within the arboretum. We monitored transpiration, leaf phenology and the chemical composition of the xylem sap of girdled and nongirdled trees. Spectral analysis revealed slightly higher amounts of sugars, especially saccharose, in Cladrastis Raf. as the most regularly girdled tree among other conditions, comparing girdled to non-girdled trees. Higher transpiration rates were not confirmed in connection with girdling—quite the opposite—Cladrastis Raf. as the most highly favored tree for girdling showed the lowest transpiration rates (in average 6 kg water per day within spring months) compared to other non-girdled trees. We presume that the birds do not choose a particular tree on the basis of any visible or chemical traits but they examine many trees within their territory. Afterwards they probably remember the position of trees whose xylem sap starts to flow early in the spring compared to other trees, as their transpiration stream is enriched with sweet organic substances that represent an advantage for the forthcoming nesting period.     

Nitrogen and methyl jasmonate induction of soybean vegetative storage protein genes

Vegetative storage protein (VSP) and VSP mRNA levels in soybean (Glycine max) leaves correlated with the amount of NH₄NO₃ provided to nonnodulated plants. The mRNA level declined as leaves matured, but high levels of N delayed the decline. This is consistent with the proposed role for VSP in the temporary storage of N. Wounding, petiole girdling, and treatment with methyljasmonate (MeJA) increased VSP mRNA in leaves 24 hours after treatment. The magnitude of the response depended on leaf age and N availability. N deficiency essentially eliminated the response to wounding and petiole girdling. MeJA was almost as effective in N-deficient plants as in those receiving abundant N. Inhibitors of lipoxygenase, the first enzyme in the jasmonic acid biosynthetic pathway, blocked induction by wounding and petiole girdling but not by MeJA. This supports a role for endogenous leaf jasmonic acid (or MeJA) in the regulation of VSP gene expression.     

Xylem structure of four grape varieties and 12 alternative hosts to the xylem-limited bacterium Xylella fastidious

Background and Aims

The bacterium Xylella fastidiosa (Xf), responsible for Pierce''s disease (PD) of grapevine, colonizes the xylem conduits of vines, ultimately killing the plant. However, Vitis vinifera grapevine varieties differ in their susceptibility to Xf and numerous other plant species tolerate Xf populations without showing symptoms. The aim of this study was to examine the xylem structure of grapevines with different susceptibilities to Xf infection, as well as the xylem structure of non-grape plant species that support or limit movement of Xf to determine if anatomical differences might explain some of the differences in susceptibility to Xf.

Methods

Air and paint were introduced into leaves and stems to examine the connectivity between stem and leaves and the length distribution of their vessels. Leaf petiole and stem anatomies were studied to determine the basis for the free or restricted movement of Xf into the plant.

Key Results

There were no obvious differences in stem or petiole vascular anatomy among the grape varieties examined, nor among the other plant species that would explain differences in resistance to Xf. Among grape varieties, the more tolerant ‘Sylvaner’ had smaller stem vessel diameters and 20 % more parenchyma rays than the other three varieties. Alternative hosts supporting Xf movement had slightly longer open xylem conduits within leaves, and more connection between stem and leaves, when compared with alternative hosts that limit Xf movement.

Conclusions

Stem–leaf connectivity via open xylem conduits and vessel length is not responsible for differences in PD tolerance among grape varieties, or for limiting bacterial movement in the tolerant plant species. However, it was found that tolerant host plants had narrower vessels and more parenchyma rays, possibly restricting bacterial movement at the level of the vessels. The implications of xylem structure and connectivity for the means and regulation of bacterial movement are discussed.     

Are leaves ‘freewheelin'? Testing for a Wheeler‐type effect in leaf xylem hydraulic decline

A recent study found that cutting shoots under water while xylem was under tension (which has been the standard protocol for the past few decades) could produce artefactual embolisms inside the xylem, overestimating hydraulic vulnerability relative to shoots cut under water after relaxing xylem tension (Wheeler et al. 2013). That study also raised the possibility that such a ‘Wheeler effect’ might occur in studies of leaf hydraulic vulnerability. We tested for such an effect for four species by applying a modified vacuum pump method to leaves with minor veins severed, to construct leaf xylem hydraulic vulnerability curves. We tested for an impact on leaf xylem hydraulic conductance (K_x) of cutting the petiole and minor veins under water for dehydrated leaves with xylem under tension compared with dehydrated leaves after previously relaxing xylem tension. Our results showed no significant ‘cutting artefact’ for leaf xylem. The lack of an effect for leaves could not be explained by narrower or shorter xylem conduits, and may be due to lesser mechanical stress imposed when cutting leaf petioles, and/or to rapid refilling of emboli in petioles. These findings provide the first validation of previous measurements of leaf hydraulic vulnerability against this potential artefact.     

Impact of phloem girdling on leaf gas exchange and hydraulic conductance in hybrid aspen

We investigated phloem-xylem interactions in relation to leaf hydraulic capacity in hybrid aspen (Populus tremula L. × P. tremuloides Michx.) by using phloem girdling method. Removal of bark tissues (phloem girdling) at the branch base resulted in a substantial decline in stomatal conductance (g_S), net photosynthetic rate (P_N), and leaf hydraulic efficiency, and in increase of leaf water potential (Ψ_L). Although gS declined more than P_N (83 versus 78 %), the ratio of intercellular to ambient CO₂ concentrations (c_i/c_a) increased from 0.67 to 0.87 in three days after girdling. Girdling induced a decrease in leaf hydraulic conductance (K_L) on average by 43 % (P = 0.006). The changes in gS and leaf conductance to water vapour were co-ordinated with K_L only in girdled branches whereas intrinsic water-use efficiency was invariant to K_L. The declines in K_L with girdling were not accompanied by changes in potassium ion concentration ([K⁺]), electrical conductivity, or pH of xylem sap. The results suggest that phloem girdling at the branch base does not influence the recirculation of ions between the phloem and xylem in hybrid aspen and the decrease of K_L in response to the manipulation is not related to changes in [K⁺] and total ionic content of xylem sap.     

Decline in gypsy moth (Lymantria dispar) performance in an elevated CO2 atmosphere depends upon host plant species

Plant species differ broadly in their responses to an elevated CO₂ atmosphere, particularly in the extent of nitrogen dilution of leaf tissue. Insect herbivores are often limited by the availability of nutrients, such as nitrogen, in their host plant tissue and may therefore respond differentially on different plant species grown in CO₂-enriched environments. We reared gyspy moth larvae (Lymantria dispar) in situ on seedlings of yellow birch (Betula allegheniensis) and gray birch (B. populifolia) grown in an ambient (350 ppm) or elevated (700 ppm) CO₂ atmosphere to test whether larval responses in the elevated CO₂ atmosphere were species-dependent. We report that female gypsy moths (Lymantria dispar) reared on gray birch (Betula populifolia) achieved similar pupal masses on plants grown at an ambient or an elevated CO₂ concentration. However, on yellow birch (B. allegheniensis), female pupal mass was 38% smaller on plants in the elevated-CO₂ atmosphere. Larval mortality was significantly higher on yellow birch than gray birch, but did not differ between the CO₂ treatments. Relative growth rate declined more in the elevated CO₂ atmosphere for larvae on yellow birch than for those on gray birch. In preference tests, larvae preferred ambient over elevated CO₂-grown leaves of yellow birch, but showed no preference between gray birch leaves from the two CO₂ atmospheres. This differential response of gypsy moths to their host species corresponded to a greater decline in leaf nutritional quality in the elevated CO₂ atmosphere in yellow birch than in gray birch. Leaf nitrogen content of yellow birch dropped from 2.68% to 1.99% while that of gray birch leaves only declined from 3.23% to 2.63%. Meanwhile, leaf condensed tannin concentration increased from 8.92% to 11.45% in yellow birch leaves while gray birch leaves only increased from 10.72% to 12.34%. Thus the declines in larval performance in a future atmosphere may be substantial and host-species-specific.     

Comparison of water ingestion between black flies (Simuliidae) and a mosquito (Aedes aegypti; Culicidae) using radiolabelled glucose

Ingestion of water by larvae of the black flies Simulium verecundum, Prosimulium mixtum/fuscum and Cnephnia dacotensis and by larvae of the mosquito Aedes aegypti was studied with radiolabelled glucose as a tracer. Extremely low ( 200 dpm/3 h) radiolabel uptake occurred in the black flies species, while Aedes aegypti, a species that is known to drink, had high radiolabel counts (> 6000 dpm/3 h). Radiolabel uptake by S. verecundum did not change for larvae fed in absence of current or with plugged mouthparts versus normal larvae. Radiolabel uptake by Aedes aegypti larvae with plugged mouthparts was < 3% of radiolabel uptake in normal larvae. Aedes aegypti was able to ingest the soluble glucose. Simulium verecundum, Prosimulium mixtum/fuscum and Cnephia dacotensis were unable to ingest the soluble sugar. The rationale for this difference is in the different filter-feeding mechanism of Culicidae and Simuliidae.     

Salmonella Survival on Pecans as Influenced by Processing and Storage Conditions

Survival of Salmonella senftenberg 775W, S. anatum, and S. typhimurium during exposure to currently practiced, as well as abusive, pecan processing and storage conditions was studied. Thermal treatments normally carried out during the processing of pecans are inadequate to consistently destory salmonellae in highly contaminated inshell nuts. Pecan nut packing tissue was toxic to salmonellae, thus affording some protection against high initial contamination and subsequent survival of the organisms. Examinations of inoculated inshell pecans stored at -18, -7, 5, and 21 C for up to 32 weeks revealed that the extent of survival was inversely correlated to the storage temperature. S. senftenberg 775W and S. anatum were not detectable on inshell nuts after 16 weeks of storage at 21 C. Little decrease in viable population of the three species was noted on inoculated pecan halves stored at -18, -7, and 5 C for 32 weeks. Due to organoleptic quality deterioration in pecan nutmeats at elevated temperatures, sterilization methods other than thermal treatment appear to be required for the elimination of viable salmonellae from pecan nuts.     

Variations in leaf characteristics of six species of Sagittaria (alismataceae) caused by various water levels

Plants grown from seeds of Sagittaria falcata, S. lancifolia, S. platyphylla, S. rigida, S. isoetiformis and S. papillosa were grown in water-saturated soil or soil submerged to 4.5, 12, 19.5 or 27 cm. Length and width of leaves and petiole lengths were measured at anthesis of the first flower on the first inflorescence produced by each plant. In general, leaf width and length were decreased by submergence, and petiole length increased. The species × water depth interaction was significant for emersed leaf width, leaf length and petiole length except for S. lancifolia leaf length and S. papillosa leaf length and width. These trends indicate genetic differences among, and variability within, taxa. Leaf width, leaf length and petiole length of plants growing in seed source populations were measured. The means from these measurements, when compared to those from experimental plants, indicate that both groups of plants respond similarly to variations in water depth.     

Larval recognition by Temnothorax longispinosus and T. ambiguus hosts of the slave-making ant Protomognathus americanus revisited: colony-level referent ensures conspecific preference

Slave-making ants exploit the labour of their own or another species. Temnothorax ambiguus and T. longispinosus are both ant species that serve as hosts of the obligatory slave-making ant Protomognathus americanus and are facultative slave-makers themselves. We offered laboratory colonies of T. ambiguus and T. longispinosus a series of choices among different larval types to better understand their brood discrimination abilities. Workers of both species preferentially accepted nestmate over non-nestmate larvae. Both species preferentially retrieved unrelated conspecific larvae over congeneric allospecific larvae, and T. ambiguus workers consumed more allospecifics than conspecifics. When presented with conspecific versus P. americanus larvae, both species manifested a clear bias towards conspecific larvae in terms of earlier retrieval and reduced cannibalism. That workers did not prefer P. americanus larvae over conspecific larvae as documented in previous research likely reflects the fact that in the present study, subject workers had access to the entirety of their colony as a referent during rearing and at the time of testing, as they would in nature. Moreover, reciprocal contact between P. americanus and conspecific larvae increased acceptance of the slave-maker larvae, but did not appear to lessen the acceptability of conspecific larvae. This suggests that transfer of cues through contact may be sufficient to alter the recognition signature of P. americanus larvae increasing acceptability by their hosts.     

Mouthparts of larvae of Opifex fuscus and Aedes australis (Diptera: Gulicidae); a scanning electron microscope study

Abstract

Larvae of the New Zealand culicid species Opifex fuscus and Aedes australis have previously been reported to show dimorphism in the structure of their labral brushes, some larvae having pectinate bristles and others only simple hairs. In the scanning electron microscope all larvae showed some degree of pectination of hairs in these brushes. There is also a gradation in the pectination. Some bristles are only sparsely pectinate; because the dimensions of their teeth are close to the limit of resolution by the compound microscope, the pectination had previously gone undetected. The mouthparts of both species are intermediate in character between those typical of filter-feeding larvae and those typical of browsing larvae. The SEM appearance of maxillary sensoria and bristles on the ventral surfaces of the mandibular brushes is described; the latter bristles comb food particles out of the labral brushes and towards the mouth. Features of the mouthparts are illustrated with scanning electron micrographs.     

Gelechiidae moths are capable of chemically dissolving the pollen of their host plants: first documented sporopollenin breakdown by an animal

Background

Many insects feed on pollen surface lipids and contents accessible through the germination pores. Pollen walls, however, are not broken down because they consist of sporopollenin and are highly resistant to physical and enzymatic damage. Here we report that certain Microlepidoptera chemically dissolve pollen grains with exudates from their mouthparts.

Methodology/Principal Findings

Field observations and experiments in tropical China revealed that two species of Deltophora (Gelechioidea) are the exclusive pollinators of two species of Phyllanthus (Phyllanthaceae) on which their larvae develop and from which the adults take pollen and nectar. DNA sequences placed the moths and plants phylogenetically and confirmed that larvae were those of the pollinating moths; molecular clock dating suggests that the moth clade is younger than the plant clade. Captive moths with pollen on their mouthparts after 2-3 days of starvation no longer carried intact grains, and SEM photographs showed exine fragments on their proboscises. GC-MS revealed cis-β-ocimene as the dominant volatile in leaves and flowers, but GC-MS analyses of proboscis extracts failed to reveal an obvious sporopollenin-dissolving compound. A candidate is ethanolamine, which occurs in insect hemolymphs and is used to dissolve sporopollenin by palynologists.

Conclusions/Significance

This is the first report of any insect and indeed any animal chemically dissolving pollen.     

Cytokinins in the Phloem Sap of White Lupin (Lupinus albus L.)

Cytokinin-like activity in samples of xylem and phloem sap collected from field-grown plants of white lupin (Lupinus albus L.) over a period of 9 to 24 weeks after sowing was measured using the soybean hypocotyl callus bioassay following paper chromatographic separation. The phloem sap was collected from shallow incisions made at the base of the stem, the base of the inflorescence (e.g. stem top), the petioles, and the base and tip of the fruit. Xylem sap was collected as root exudate from the stump of plants severed a few centimeters above ground level. Concentration of cytokinin-like substances was highest in phloem sap collected from the base of the inflorescence and showed an increase over the entire sampling period (from week 10 [61 nanogram zeatin equivalents] to week 24 [407 nanogram zeatin equivalents]). Concentrations in the xylem sap and in the other phloem saps were generally lower. Relatively high concentrations of cytokinin-like substances in petiole phloem sap (70 to 130 nanogram zeatin equivalents per milliliter) coincided in time with high concentrations in sap from the base of the inflorescence (see above). Concentrations in sap (phloem or xylem) from the base of the stem were very much lower. This finding is consistent with movement of cytokinins from leaves into the developing inflorescence and fruit, rather than direct input to the fruit from xylem sap. However, an earlier movement of cytokinins from roots into leaves via the xylem cannot be ruled out. Sap collected at an 18-week harvest was additionally separated by sequential C₁₈ reversed-phase high performance liquid chromatography → NH₂ normal phase high performance liquid chromatography, bioassayed, and then analyzed by electron impact gas chromatography-mass spectrometry. Identification of zeatin riboside and dihydrozeatin as two of the major cytokinins in combined sap samples was accomplished by gas chromatography-mass spectrometry-selected ion monitoring.     

Recycling Potential and Fitness of Steinernematid Nematodes Cultured in Curculio caryae and Galleria mellonella

The recycling potential of entomopathogenic nematodes in the pecan weevil, Curculio caryae, following inundative applications is an important factor in considering whether nematodes could be incorporated into a C. caryae management strategy. Our objective was to determine the recycling potential and fitness of Steinernema carpocapsae and S. riobrave cultured in C. caryae. To estimate fitness and quality, we reared nematodes in larvae of C. caryae and in the commonly used standard host, the greater wax moth, Galleria mellonella. Nematode lipid content, infectivity (power to invade), virulence (power to kill), and reproductive capacity (yield per insect) in C. caryae larvae were compared with G. mellonella data. Lipid content was higher in S. carpocapsae cultured in C. caryae than in G. mellonella, but S. riobrave lipid content was not affected by host source. Host source did not affect subsequent infectivity or virulence to C. caryae (P > 0.05) but did affect reproductive capacity (P < 0.0001). Both nematode species produced more progeny in C. caryae when they were first cultured in G. mellonella than when they were first passed through C. caryae. In terms of potential to recycle under field conditions, we predict that nematodes resulting from one round of recycling in C. caryae larvae would be equally capable of infecting and killing more weevils, but the potential to continue recycling in C. caryae would diminish over time due to reduced reproduction in that host.     

Distribution of the black pecan aphid,Melanocallis caryaefoliae,on the upper and lower surface of pecan foliage

Three aphid species regularly feed on pecan [Carya illinoinensis (Wangenh.) K. Koch (Juglandaceae)] foliage: the black pecan aphid, Melanocallis caryaefoliae (Davis), the yellow pecan aphid, Monelliopsis pecanis Bissell, and the blackmargined aphid, Monellia caryella (Fitch) (all Hemiptera: Aphididae). Adults of M. caryaefoliae and both the nymphs and adults of M. pecanis and M. caryella mainly feed on the lower surface of leaves. Nymphs of M. caryaefoliae appear unique by frequently feeding on the upper surface of pecan leaves. This is risky behavior given the environmental hazards (e.g., rain, solar radiation, and dislodgement) associated with the upper surface. Thus, we determined the leaf surface distribution of M. caryaefoliae on trees in an orchard and on pecan seedlings in the laboratory. A pecan orchard survey found all three aphid species and stages predominantly on the lower leaf surface, except for the nymphs of M. caryaefoliae, which were evenly distributed between upper and lower leaf surfaces. This survey also found aphidophagous lacewing (Neuroptera) larvae predominantly on the lower leaf surface, whereas ladybird beetle (Coleoptera: Coccinellidae) larvae were more evenly distributed between upper and lower surfaces. Laboratory experiments using single or multiple pecan aphid species revealed M. caryaefoliae distribution on pecan seedlings similar to orchard data. Nymphal M. caryaefoliae require nearly 2 days to elicit chlorotic feeding lesions on leaves; without these lesions, nymphal development is hindered. The similar distribution of nymphs of M. caryaefoliae on both leaf surfaces likely reflects a strategy of predator avoidance allowing a proportion of the population to survive.