Maternal death relaxes developmental inhibition in nymphal aphid defenders

In certain aphids, first-instar nymphs defend their gall by attacking intruding arthropod predators. One correlate of such defensive behaviour is a lengthened duration of the first nymphal stadium during the galling phase of the life cycle. A prolonged first stadium allows a large army of first-instar defenders to accumulate, which may be advantageous for gall defence. The factors determining developmental delay have been unclear, however. Our field experiment with Pemphigus obesinymphae, a North American gall-forming aphid with defensive first-instar nymphs, tests whether first-stadium duration is influenced by the death of the colony''s fundatrix (mother). We killed fundatrices in certain galls, left those in control galls alive, and counted aphids in each stadium in each gall. Galls in which fundatrices were killed contained a lower proportion of first-instar defenders and more late-instar nymphs than did galls with living fundatrices, indicating that maternal death dramatically increased developmental rate of nymphs. Possibly nymphal aphids respond adaptively to environmental cues that signal a threat to the colony''s welfare. Alternatively, the fundatrix actively suppresses offspring development in order to maintain a large army of soldiers to protect her gall. The results add a new layer of complexity to our understanding of social aphid systems.     

Shift in life-history strategy from reproduction to defense with colony age in the galling aphidHemipodaphis persimilis producing defensive first-instar larvae

Colony defense in some aphids is performed by sterile soldiers but in others by monomorphic larvae of a specific instar stage. This paper, focusing on a galling aphid Hemipodaphis persimilis with monomorphic defensive first instars, examined the mechanism by which the proportion of defenders is regulated in the colonies. Demographic analyses showed that the ratios of first instars (defenders) were kept constantly high (58% on average) from mid June to late September. High proportions of first instars could be explained by consistently high birth rates (birth rate hypothesis) or by a prolonged duration of the first instar stage (instar span hypothesis). With the progress of colony age, the mature-embryo content of apterous adults, used as an index of the birth rate, decreased and the proportions of advanced instars increased. These results did not support the birth rate hypothesis. By contrast, calculation of a newly proposed index, the molting rate, showed that the duration of the first-instar stage was short in incipient galls but became longer with colony age. The duration of other instar stages was always kept short. These results corroborate the instar span hypothesis and suggest that the prolongation of the first-instar stage is an adaptive mechanism by which the defender ratio is kept high in mature colonies where the birth rate is declining. The frequency of aggressive behavior in first instars increased from incipient to mature galls. Seasonal changes in the instar span and aggressiveness of first instars suggest that in H. persimilis colonies there is a strategic shift from the reproductive to defensive phase with colony age.     

Defenders in the North American aphidPemphigus obesinymphae

Summary Gall-inhabiting individuals of the aphidPemphigus obesinymphae act as defenders, protecting other colony members against attack by dipteran and neuropteran larvae that are the primary predators of this species. As first instar nymphs, the progeny of the fundatrix patrol surfaces of galls and adjoining leaves. These first instar nymphs attack potential predators by mounting and grasping them and inserting their stylets. This defensive behavior, which is not exhibited by nymphs in later instars, appears to be effective in reducing predation. The fundatrix typically produces defenders throughout the extended gall-inhabiting phase, and her progeny delay development beyond the defensive first instar stage. By August, galls contain an average of 101 defenders. Early death of the fundatrix reduces the number of defenders in the gall and advances maturation of defenders into winged migrants, which otherwise mature in September and October. InPemphigus, defensive behavior by first instar nymphs appears to have evolved in the context of several types of derived life cycle, each involving an extended gall-inhabiting phase.     

Soldier caste with morphological and reproductive division in the aphid tribe Nipponaphidini

Summary. We investigated demographic, morphological and histological aspects of Distylaphis foliorum, a gall-forming nipponaphidine aphid from Java, Indonesia, whose first instar nymphs had been reported to have enlarged forelegs and attack other insects. The gall inhabitants of D. foliorum consisted of two discrete populations; one developing normally and the other remaining at the first instar. Morphometric analysis identified two types of first instar nymphs; nymphs with relatively long forelegs and slender abdomen, and nymphs with relatively short forelegs and fat abdomen. The former nymphs were found from both young galls and mature galls, while the latter nymphs were from young galls only. Histological analysis strongly suggested a reproductive division in the first instar. In the former nymphs from mature galls, ovaries and mycetomes were degenerate and replaced by well-developed fat body cells. These results indicated that D. foliorum has a morphologically differentiated sterile soldier caste in the first instar. This study is the first report of a soldier caste with morphological and reproductive division in the aphid tribe Nipponaphidini, and suggests that highly specialized soldier castes have evolved at least four times in aphids.Received 25 June 2004; revised 24 September 2004; accepted 11 October 2004.     

Do aphid galls provide good nutrients for the aphids?: Comparisons of amino acid concentrations in galls among Tetraneura species (Aphididae: Eriosomatinae)

Some aphid species induce leaf galls, in which the fundatrix parthenogenetically produces many nymphs. In order to ensure high performance, galls have to provide the aphids with sufficient nutrients, in particular, amino acids as a nitrogen source. We tested this hypothesis using six Tetraneura aphid species that induce closed galls. We extracted free amino acids from the whole gall tissues of unit weight and quantified the concentration of each amino acid. There were large differences in the total amino acid concentrations among galls of the Tetraneura species. Tetraneura species in which higher concentrations of total amino acids were found in the gall tended to produce larger numbers of offspring. Of the amino acids found, asparagine was predominant in the gall. The asparagine concentration in T. yezoensis galls was several hundred times as high as in control leaves. We discussed why such a high level of asparagine accumulates in aphid galls.     

Formation of long‐lasting galls by overwintered nymphs in the Japanese aphid Quadrartus yoshinomiyai (Hemiptera: Aphididae: Hormaphidinae)

This paper examines the life history of a generation of galls created by the aphid Quadrartus yoshinomiyai (Hormaphidinae: Nipponaphidini) on its primary host plant, Distylium racemosum. First‐instar fundatrix nymphs of Q. yoshinomiyai initiated galls on stems of developing shoots in early April and incipient enclosed galls were found from later the same month. The galls lasted for up to 14 months, during which they grew to maturity, opened in early or mid‐April of the following year and dried up by the end of June. First‐instar fundatrix nymphs were found on winter buds, indicating that they hatched from eggs in autumn and overwintered as nymphs. These results suggest that Q. yoshinomiyai has a three‐year life cycle.     

Defense by a few first-instar nymphs in the closed gall ofdinipponaphis autumna (homoptera, aphididae, hormaphidinae)

The defensive behavior of the aphidDinipponaphis autumna, which forms small, completely closed galls on leaves ofDistylium racemosum, was observed. In mature galls, in addition to tens of mature or nearly mature aphids of the 3rd generation, there remained 3 – 17 1st-instar nymphs of the same generation, which had well-sclerotized legs and the antennae with developed setae. Despite their minority, these 1st-instar nymphs clung to experimentally introduced insect larvae and stung them with their stylets. Fourth-instar wingpadded nymphs, the majority of the 3rd generation at the experiment, also attacked the introduced larvae, but they were readily spilt from the larvae. Four out of 205 1st-instar nymphs remaining in mature galls had the next instar cuticle developing inside, indicating that they are not destined to be sterile.     

Defensive Behavior in Primary- and Secondary-Host Generations of the Soldier-Producing Aphid, Pemphigus bursarius (Hemiptera: Aphididae)

The genus Pemphigus comprises several species that produce soldiers (defensive morphs) in galls on the primary host. At the moment, it is unclear if host-alternating species also produce defenders on their secondary host. We therefore examined how P. bursarius morphs of the secondary host generations respond to predators to test whether they show defensive behavior. We further examined how this response compares with the antipredator behavior of soldiers in P. bursarius and P. spyrothecae occurring on the primary host. We performed two manipulative experiments using two predatory species to quantify the behavior of the different morphs in response to predators. In both experiments, secondary host morphs of P. bursarius showed no attacking behavior and antipredator behavior in these morphs was limited to escaping natural enemies by walking away. In contrast, the first instars of the primary host generations in both Pemphigus species showed attacking behavior and were capable of killing the predators.     

Tuberaphis owadai (Homoptera), a new aphid species forming a large gall on Styrax tonkinensis in northern Vietnam

Tuberaphis owadai sp. nov., an aphid species forming coral‐shaped galls on Styrax tonkinensis in northern Vietnam, is described. We found that the species produces many sterile second‐instar soldiers in the gall. The colony size of a large gall was estimated to be 178 000, approximately half of which were soldiers. Alates emerging from galls contained sexual embryos, indicating that the life cycle is monoecious (non‐host‐alternating). Predaceous larvae of the pyralid moth Assara seminivalis were found in several galls.     

The effects of host-plant properties on gall density,gall weight and clone size in the aphidAploneura lentisci (Pass.) (Aphididae: Fordinae) in Israel

Summary Samples of shoots ofPistacia lentiscus carrying galls of the aphid,Aploneura lentisci, were collected at three localities in Israel. Shoots growing near pruning scars carried more galls than elsewhere on the plant, but these galls weighed less and contained fewer aphids (smaller clones). The proportion of empty galls increased with gall density. Crowding of galls at such sites may be due to the early burst of buds at the time of aphid emergence from the overwintering eggs, and not to active search for preferred sites. Shoots bearing larger numbers of leaves carried heavier galls, which contained larger aphid clones. The position of the galled leaf on the shoot had no effect on gall weight nor on clone size. The physiological condition of the plant may be an important environmental (ecological) factor affecting the variation in clone-size and in aphid morphology among galls.     

First incidence of inquilinism in gall-forming psyllids, with a description of the new inquiline species (Insecta, Hemiptera, Psylloidea, Psyllidae, Spondyliaspidinae)

The two largest lineages of holometabolous gall-forming insects, cynipid wasps and cecidomyiid flies, have given rise to numerous obligate inquilines, species which are unable to form galls themselves and survive by inhabiting galls formed by other species. In contrast, only a single obligate inquiline, an aphid, is known in the sternorrhynchous Hemiptera, the hemimetabolan lineage in which gall-forming is best developed. We describe the first known gall inquiline in psyllids (Sternorrhyncha, Psylloidea), Pachypsylla cohabitans Yang & Riemann sp. n. All other members of this genus produce closed galls on hackberries, Celtis spp. (Ulmaceae). Newly hatched nymphs of P. cohabitans feed next to nymphs of several species of leaf gall-makers, becoming incorporated into the gall as the stationary nymphs are gradually enveloped by leaf tissue. In the mature gall, the inquilines occupy separate, lateral cells surrounding a central cell containing a single gall-maker. Pachypsylla cohabitans is similar in morphology to leaf-gallers, but differs in nymphal and adult colour, allozyme frequency, especially in the malic enzyme, and in adult phenology. Laboratory-reared progeny of side-cell females, when caged alone, never form galls, while progeny of centre-cell individuals alone only form galls comprising single individuals. Multiple-cell galls are formed only when adults of side-cell and centre-cell individuals are caged together. Experimental removal of centre-cell nymphs in early stages of gall initiation leads to smaller galls or death of side-cell individuals. We conclude that the side-cell individual is an obligate inquiline that is incapable of forming a gall on its own but is derived from a leaf-galling ancestor. We speculate on selective forces that might favour this evolutionary transition.     

Secondary host generation of the gall aphid Cerataphis jamuritsu (Homoptera)

Colonies of a Cerataphis species with well‐developed horns were found on the rattan Calamus quinquesstinervis in southern Taiwan. The morphology of first instar nymphs from the colonies accorded well with the morphology of first instar nymphs laid by alates of Cerataphis jamuritsu from galls on Styrax suberifolia, indicating that the rattan aphids are the secondary host generation of C. jamuritsu. Although the aphid colonies were attended by ants, the sharp horns of the first instar nymphs suggest that they might attack predators.     

Seasonal extension of the soldier instar as a route to increased defence investment in the social aphid Pemphigus spyrothecae

Abstract.  1. The possibility that the duration of the soldier instar could be extended over the course of the galling season was examined in Pemphigus spyrothecae Passerini (Hemiptera: Aphididae). In this species, the individuals of one morph of the dimorphic first instar are non-sterile specialised defenders known as soldiers.
2. In the face of a declining birth rate, the proportion of first-instar soldiers to first-instar non-soldiers was found to increase. This provides strong evidence of extension of the soldier instar over time.
3. Instar extension provides a mechanism for post-natal investment in soldiers and allows individuals with insufficient time to mature to remain in the defensive stage so that they may continue to make an indirect contribution to their clone's reproductive success.     

Clonal mixing in the soldier-producing aphid Pemphigus spyrothecae (Hemiptera: Aphididae)

Illuminating the genetic relationships within soldier-producing aphid colonies is an essential element of any attempt to explain the evolution of the altruistic soldier caste. Pemphigus spyrothecae is a soldier-producing aphid that induces galls on the leaf petioles of its host (trees of the genus Populus). At least a quarter of the aphids within the clonally produced gall population are morphologically and behaviourally distinct first-instar soldiers that defend the gall population from predation. Using field trapping and microsatellites, we investigated the degree of clonal mixing within natural gall populations. Field trapping in the UK showed that all the migrants of P. spyrothecae and of two other Pemphigus species were wingless first-instar soldiers. The average degree of mixing estimated from trapping P. spyrothecae migrants was 0.68% (range = 0-15%). Microsatellite genotyping of 277 aphids from 13 galls collected in Italy revealed an average mixing level of 10.4% (range = 0-59%). Six galls contained more than one clone (range = 2-5 clones). Non-kin aphids were not restricted to the soldier caste but were evenly distributed across instars. An additional gall, from which 527 occupants were genotyped, contained 12 non-kin aphids distributed among nine clones, showing that clonal diversity can be high even when mixing is very low. These observations suggest that although soldiers migrate regularly and can moult and reproduce within foreign galls, clonal mixing in this species is generally low and is unlikely to provide a barrier to the evolution of investment by the aphid clones in an altruistic soldier caste.     

Comparative analysis of caste differentiation during embryogenesis of social aphids whose soldier castes evolved independently

Summary. We investigated the morphological characters of normal nymphs, soldier nymphs and developing embryos of a social aphid, Colophina arma, which has a sterile soldier caste in the first instar. Morphometric analysis revealed that normal nymphs and soldier nymphs were clearly distinguishable on the basis of several morphological characters. At late embryonic stages, normal embryos and soldier embryos were also distinguishable morphologically. The younger the embryonic stages, the smaller the morphological differences between them. In young embryos of less than 600 m in body length, normal embryos and soldier embryos were no more distinguishable, suggesting that the onset of soldier differentiation occurs at an early embryonic stage. Through the embryonic development, morphological differentiation of soldier caste proceeded gradually: forelegs and midlegs were exaggerated, and growth of mouthpart was suppressed, which resulted in the soldier morphology specialized for attacking behavior. On the basis of these results, developmental aspects in soldier differentiation of C. arma were compared with those of Pseudoregma bambucicola, a social aphid with a first instar soldier caste of independent evolutionary origin. Ecological and evolutionary relevance of the differences between the two social aphids was discussed.Received 30 June 2004; revised 20 October 2004; accepted 9 November 2004.     

The relationship between conidial dose,moulting and insect developmental stage on the susceptibility of cotton aphid,Aphis gossypii,to conidia of Lecanicillium attenuatum,an entomopathogenic fungus

The cotton aphid is one of the most serious pests of greenhouse vegetable crops worldwide. It is difficult to control because field populations usually include simultaneously several insect developmental stages. The current research evaluated an isolate (CS625) of Lecanicillium attenuatum, a fungal pathogen of aphids, as to its virulence against different developmental stages of cotton aphid, Aphis gossypii. The influence on mortality of several other factors also was examined: (a) insect moulting, (b) the number of conidia attached to insect cuticles and (c) germination rates of conidia on cuticles of aphids at various developmental stages. Mortality of cotton aphids treated with L. attenuatum conidia varied according to the developmental stage of the host, i.e. the LT_50s with third-instar nymphs and adults was shorter than with first-instar nymphs. The number of spores attached to the surface of first-instar nymphs was approximately one-half of that on third-instar nymphs and adults. Also, the level of spore germination on the surface of first-instar nymphs was lower than on the surface of other stages of the aphid. After moulting, the numbers of conidia attached to new insect cuticles were less than on exuviae. These results suggest that early nymphal stages of cotton aphids may escape fungal disease due, at least in part, to a combination of three factors: low numbers of conidia attached to their cuticles; low levels of conidial germination and rapid ecdyses, which removed conidia before their germ tubes penetrated the host hemolymph.     

Effects of a juvenile hormone analogue, pyriproxyfen, on the apterous form of soybean aphid (Aphis glycines)

The soybean aphid, Aphis glycines Matsamura, is an eastern Asian soybean [Glycine max (L.) Merr.] pest which can reduce soybean yield. We determined the effects of pyriproxyfen, a juvenile hormone analogue, on development, mortality, longevity and fecundity of A. glycines under laboratory conditions. Distance Insect Growth Regulator, containing ∼11.2% pyriproxyfen, was applied at two concentrations, 50 and 150 mg/l, to first and fourth instar nymphs. When first or fourth instar A. glycines were treated with pyriproxyfen, some nymphs became supernumerary‐molted nymphs with 1–3 extra molts or were sterilized. Mortality of treated first instar nymphs was >68% greater than the control group and longevity was reduced by >40%. The higher concentration of pyriproxyfen reduced fecundity of first instar nymphs when they reached adulthood by ∼79%. Pyriproxyfen similarly affected fourth instar nymphs. Mortality of treated fourth instar nymphs was ≥15% greater than the control group and longevity was reduced by >24%. Both concentrations of pyriproxyfen lowered the fecundity of fourth instar nymphs by >27%. Pyriproxyfen also had other sublethal effects on fourth instar nymphs which became apparent when they molted to adulthood. In a few instances they developed wing pads and many produced dead, deformed or abnormal neonates that lacked appendages.     

High mean relatedness among communally galling Tamalia aphids revealed by AFLP analysis

Cooperative or eusocial behavior occurs in gall-inducing insects, but the ecological and evolutionary contexts for these behaviors vary in their details. Foundresses of the manzanita leaf-gall aphid, Tamalia coweni, regularly share galls. Because aphids undergo parthenogenesis, communal foundresses are potential clone mates. Under the kin selection hypothesis, a high level of relatedness is predicted to lower genetically based conflict among females in a group and thereby favor communal gall occupation. We recorded the frequency of communal behavior in a population of T. coweni on its host plant, Arctostaphylos patula, and measured reproductive output in single- and multiple-occupant galls. Eleven percent of the 375 galls examined were communally occupied, with double-foundress galls the commonest class, up to a maximum of five foundresses within galls. Total productivity of communal galls (measured by numbers of offspring per gall) was higher than for single-foundress galls on a per-gall basis, but lower per capita. We genotyped foundresses with amplified fragment length polymorphism (AFLP)-PCR markers, to estimate relatedness among gall co-occupants and foundresses from randomly selected galls in the population. Analysis of genetic distance between communal foundresses revealed that relatedness among gall cohabitants was significantly higher, on average, than for foundresses drawn from the population at random (P < 0.001). Phylogenetic analysis using parsimony (PAUP) of the AFLP profiles indicated that half the foundresses sharing galls were most closely related to their gall mates. Our results are consistent with kin selection theory, and suggest that communal gall occupation in this species may be interpreted as cooperative behavior.     

On the pronymphal stage in the migratory locust (Locusta migratoria, Orthoptera, Acrididae)

The structure of the integument, somatic and visceral muscles, midgut, and Malpighian tubules were investigated at the late stages of the embryonic and early postembryonic development of the migratory locust, Locusta migratoria, to assess the organization of its pronymphal stage. In its morphogenetic features, the vermiform locust larva sometimes called the pronymph corresponds to the first nymphal instar covered with the second embryonic cuticle which has not been shed. Since the first-instar locust nymphs before and after the shedding of this embryonic cuticle differ significantly in many morphological characters, two consecutive phases of this nymphal instar can be distinguished: the first phase existing from the moment of development of the third embryonic cuticle to the shedding of the second one; the second phase existing from the shedding of the second embryonic cuticle to the formation of the cuticle of the second nymphal instar. Since the pronymphal stage should precede the nymph stage, it may be concluded that the pronymph of the locust is fully embryonized and covered with the second embryonic cuticle, which is also typical of other insects with hemimetabolous development (Konopová and Zrzavý, 2005). Therefore, it would be erroneous to refer to the vermiform first-instar nymph as the pronymph, because the two stages are separated by molting and formation of a new cuticle.     

Aphid galls accumulate high concentrations of amino acids: a support for the nutrition hypothesis for gall formation

The nutrition hypothesis for the adaptive significance of insect gall formation postulates that galls accumulate higher concentrations of nutritive compounds than uninfested plant tissue, resulting in a high performance of the gall former. This hypothesis has been supported by some taxa of gall insects, but not by taxa such as cynipid wasps. Aphid galls are expected to require higher levels of nitrogen than other insects’ galls with a single inhabitant, because aphid galls are required to sustain a number of aphids reproducing parthenogenetically over two generations. The present study tested this hypothesis by evaluating aphid performance and amino acid concentration in phloem sap, using the aphid Rhopalosiphum insertum (Walker) (Homoptera: Aphididae), which establishes colonies on leaves of Sorbus commixta Hedlund or in galls of the aphid Sorbaphis chaetosiphon Shaposhnikov (Homoptera: Aphididae). We prepared the gall and non‐gall treatments on trees of S. commixta, in which R. insertum fundatrices were reared and allowed to reproduce. In S. chaetosiphon galls, R. insertum colonies propagated more rapidly, and the second generation grew larger and more fecund than on ungalled leaves. The amount of amino acids exuding from cut galled leaves was fivefold that in ungalled leaves; however, there was no significant difference in the amino acid composition between galled and ungalled leaves. In the intact leaves, total amino acid concentration in the phloem sap declined rapidly from late April to late May; however, the galls retained this high amino acid concentration in developing leaves for 1 month. These results indicate that the improved performance in R. insertum is ascribed to the increased concentration of amino acids in galled leaves. We suggest that S. chaetosiphon galls function to promote the breakdown of leaf protein, leading to an increased performance of gall‐inhabiting aphids.