Aetiology of melanized inclusions in the body cavity of Neodiprion sertifer (Geoffroy) (Hym., Diprionidae) adults

Abstract: The aetiology of large melanized inclusions occurring in the haemocoel of adults of some diprionid sawflies is not fully understood. Their occurrence has been associated with nuclear polyhedrosis virus (NPV), but also with extreme temperatures, parasitism, and prolonged diapause. High frequencies of inclusions are found most often during NPV epizootics. In laboratory experiments with Neodiprion sertifer larvae, ingestion of large doses of a common bacterium, Enterobacter cloacae , was shown to induce inclusions indistinguishable from those associated with NPV infection. It is argued that a bacterial aetiology is not contradicted by the other conditions reported to be associated with inclusions. The bacterial aetiology provides an explanation for the low frequencies of inclusions in N. sertifer populations with no NPV infection. It is proposed that the ability to form inclusions may have evolved as a defence against bacterial infection.     

Responses of Neodiprion sertifer (Hym., Diprionidae) larvae to variation in needle resin acid concentration in Scots pine

Summary We have studied how differences in needle resin acid concentrations between two clones of Scots pine influenced larval survival, larval developmental time, and cocoon weight in Neodiprion sertifer. Larvae were reared under controlled conditions in the laboratory on needles showing a three-fold difference, in resin acid concentration. Larval developmental time was significantly longer for larvae fed needles high in resin acids. No significant differences were found in cocoon weight. Larvae fed needles high in resin acids suffered significantly higher mortality, most of which occurred, in the first two instars. Data on relative growth rate for last-instar larvae, based on a traditional growth efficiency experiment, did not show any negative effects of resin acids, indicating that a possible induction takes place during early instars. Observations on the feeding behaviour of last-instar larvae, the high consumption of bark tissue (with high concentrations of resin acids) by larvae fed needles low in resin acids and preference for needle tissues high in resin acids, indicate that these larvae might actively search for tissues rich in resin acids. We conclude that the physiology and behaviour of N. sertifer has been shaped, in part, by the conflict between the negative effects of resin acids, primarily in early instars (longer developmental time with a corresponding increase in exposure to natural enemies and an increase in direct mortality), and the positive effect of resin acids in later instars (a need for resin acids for use in its own predator defence secretion).     

Areal distribution of a Neodiprion sertifer (Hym., Diprionidae) outbreak on Scots pine as related to stand condition

Analyses of a recent N. sertifer outbreak in southern Sweden were made during on-going attack, comprising a regional and local mapping of defoliation severity, as well as a study of intensity of larval consumption, abundance of cocoons and parasitation of larvae in relation to fertilizer application. It was found that (1) the outbreak was preceded by summer and autumn droughts, (2) it was limited to infertile soils, (3) pine stands aged 20–40 yr suffered most, (4) it was less severe in trees benefiting from a reduced inter-tree competition, as measured from tree-ring growth, and (5) less severe in a fertilized stand. The findings support the hypothesis on a fundamental effect of stand nutritive state on changes in number of the sawfly.     

Interspecific competition between the principal larval parasitoids of the pine sawfly,Neodiprion sertifer (Geoff.) (Hym.: Diprionidae)

Summary Interspecific competition between the 3 principal larval parasitoids of the pine sawfly, Neodiprion sertifer, is of common occurrence when total larval parasitism, and hence multiple attack rates, are high. At the intrinsic level, the ectoparasitoid Exenterus abruptorius is superior to the 2 endoparasitoids, Lophyroplectus luteator and Lamachus eques, respectively, whereas L. luteator is superior to L. eques only. During mass outbreaks of the host competition between Exenterus and Lophyroplectus is most intense, whilst Lamachus fails to build up in the presence of its 2 competitors (Fig. 1). However, due to its superior host finding ability, Lamachus dominates in areas of low host densities where the 2 other species are less efficient (Table 1). L. luteator frequently sustains heavy losses in direct competition with E. abruptorius, but these can be largely compensated in the next generation because its fertility is about 5-times higher than that of Exenterus.     

Utilization of dry matter and bioelements in larvae of Neodiprion sertifer Geoffr. (Hym., Diprionidae) feeding on Scots pine (Pinus sylvestris L.)

Summary In the laboratory the consumption (C), egestion (F), assimilation (A) and assimilation efficiency (A/C) of dry matter and of the bioelements C, N, P, K, Ca, Mg, S and Na were studied weekly during the whole feeding stage. The larvae were reared in clusters of 10 individuals (on average 3.5 and 6.5 ), fed one-year-old needles. The average values of (C), (F) and (A) were 2,950, 2,530 and 414 mg dw per cluster, respectively, implying a dry matter assimilation efficiency of only 14.0%. The mean production of tissue (P) per cluster amounted to 250 mg dw. The concentration of bioelements in faeces (in order as above) was 51.2, 0.80, 0.08, 0.36, 0.41, 0.11, 0.10 and 0.02% of dw, and in larval tissue 57.5, 7.60, 0.64, 0.85, 0.24, 0.22, 0.24 and 0.06% of dw, respectively. The concentration of N and P in larval tissue was 5.8 and 4.9 times higher than in food, that of K, Mg, S and Na about twice, that of C and Ca about equal or less. The assimilation efficiency of the bioelements was estimated at 14.5, 59.3, 47.4, 20.9, 4.6, 12.5, 13.8 and 44.4%, and the energetics (C), (F), (A), (P) and (R=A-P) at 61,900, 53,600, 8,320, 6,710 and 1,610 J per cluster, respectively. It is argued that in N. sertifer the high consumption rate, low efficiency of dry matter assimilation and low nitrogen accumulation, in all attained at a low maintenance cost, are adaptations to a food low in nitrogen content (1.3% of dw). The study is part of an investigation on the interaction between insect consumption and pine growth, performed within the Swedish Coniferous Forest Project.The authors are given in alphabetic order; for further information see Acknowledgements     

Sex pheromone perception in male pine sawflies,Neodiprion sertifer (Hymenoptera; Diprionidae)

Summary Electroantennographic and single sensillum recordings were performed on male pine sawfly, Neodiprion sertifer, antennae. Responses to the sex pheromone component (2S, 3S, 7S)- 3,7-dimethyl-2-pentadecenyl (diprionyl) acetate (SSS:OAc), to the behavioral inhibitor (2S, 3R, 7R)-diprionyl acetate (SRR:OAc), to the six other enantiomers of diprionyl acetate, and to the biosynthetic precursor diprionol were recorded. Responses to trans-perillenal, a monoterpene identified in female gland extracts and to (2S, 3S, 7S)-diprionyl propionate (SSS:OPr), a field attractant for N. sertifer and some related sawfly species were also recorded.EAG recordings demonstrated a high antennal sensitivity to SSS:OAc and to SSS:OPr. A somewhat lower response was elicited by SRR:OAc.Single sensillum recordings revealed 8–12 different cells firing in each sensillum, corresponding to the number of cells observed in earlier morphological investigations. Out of these cells all, except one, responded to SSS:OAc and to SSS:OPr. No differences in the response to the two components could be observed. The largest amplitude cell in each sensillum was specifically tuned to the behavioral antagonist, SRR:OAc. The pheromone perception system encountered in male pine sawflies thus differs clearly from that observed in moths.Abbreviation EAG electroantennogram - OAc acetate - OPr propionate     

Mating disruption of the pine sawfly Neodiprion sertifer (Hymenoptera: Diprionidae) in isolated pine stands

• 1 Previous applications of the mating disruption technique to Neodiprion sertifer resulted in decreased numbers of males caught in the treated area but no effect on sex ratio or overall population density.
• 2 The present study assessed the efficacy of mating disruption against N. sertifer outbreaks in pine stands surrounded by agricultural areas or pasture, and therefore isolated from other infested areas.
• 3 Pine stands were treated by placing dispensers with an erythro‐mixture of the acetate ester of 3,7‐dimethyl‐2‐pentadecanol every 10 m in a grid.
• 4 The efficacy of this technique was evaluated by comparing the number of males caught in sticky traps baited with synthetic pheromone, and by comparing the sex ratio and the population density of the sawfly in the subsequent generation between treated and control pine stands.
• 5 The number of males caught within treated areas was significantly lower than in the control area.
• 6 In the treated pine stands 46% of the egg clusters resulted in male cocoons only, compared to 3% of the clusters in the control stand.
• 7 A significant reduction of the sawfly population was observed in the treated pine stands. The results contrast to the earlier mating disruption attempts with N. sertifer and can probably be ascribed to the isolation of the experimental stands.

     

Two Naturally Occurring Nuclear Polyhedrosis Virus Variants of Neodiprion sertifer Geoffr. (Hymenoptera; Diprionidae)

Two isolates of Neodiprion sertifer (Geoffr.) nuclear polyhedrosis virus from Britain and North America were compared using three biochemical techniques. Alkaline protease assays of polyhedra revealed the presence of endogenous enzyme activity in the British isolate but not in the North American isolate. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of virus particle structural polypeptides revealed only minor differences, with the exception that the North American virus was contaminated with polyhedral protein. The restriction endonucleases SalI, HindIII, and HpaII were used as a definitive method of distinguishing the two variants, with all endonucleases achieving this to a greater or lesser extent. The possible significance of all of these observations is discussed in terms of their possible influence on the registration and field application of this virus.     

Fecundity and egg viability in relation to female body size in Neodiprion sertifer (Hymenoptera: Diprionidae)

Relationships between female size and fecundity, egg viability and embryonal development were studied in Neoriprion sertifer. Sawfly females, reared from the 1st instar larvae on Scots pine needles heavily affected by industrial pollutants or on relatively unpolluted needles, were induced to lay eggs parthenogenetically in the field in Finland. Females reared on polluted needles were smaller and oviposited an average of 68.8 eggs, and those reared on unpolluted needles 79.4 eggs per female. However, the higher egg viability in the former group masked the effect of reduced female size. This compensation was primarily related to the size of the females, larger individuals producing relatively fewer viable eggs in both groups.     

Effect of Mating Disruption on Reproductive Behavior in the European Pine Sawfly, Neodiprion sertifer (Hymenoptera: Diprionidae)

Females of the European pine sawfly, Neodiprion sertifer (Geoffr.) (Hymenoptera: Diprionidae), were released and observed inside and outside (=control) of areas treated with their sex pheromone for the purpose of mating disruption. In 1992 and 1993, respectively, 0 and 2.5% of the females were observed mating in the pheromone treated area during the first day, compared with 28 and 26% in the control area. Of the females that mated, approximately 50% stayed on their twigs and oviposited, while the remaining 50% disappeared. Significantly more females disappeared from their twigs inside the treated area compared to the control area in both years. No difference was found in the proportion of unmated females ovipositing (2–12%) between the treated and the control area. Predation was responsible for much of the disappearance, but deliberate dispersal was also noted. Mating and dispersal in female N. sertifer are discussed in relation to population density and overall fitness.     

Fecundity of the European pine sawfly Neodiprion sertifer (Hymenoptera, Diprionidae) related to cyclic outbreaks: Invariance effects

At the end of ontogenesis, individual variability of the fecundity of females of Neodiprion sertifer constitutes 30–120 eggs (with rare fluctuations from 16 to 140 eggs). Average fecundity as an adaptive index in a certain biocenosis, however, depends on many factors, including the structural organization of the sawfly population. The presence of a long diapause in some individuals results in the cleavage of the offspring into groups with different dates of reactivation. At the same time, mainly heavy eonymphs, giving rise to large females, stay in the diapause. Their participation in the reproduction together with females, raised in other years under different conditions, equalize the dimensional structure of the breeding fraction of the population. During population waves, revealed during 35 years of my continuous observations, fecundity fluctuated closely to the average value with deviations constituting 19–25%. It hardly depends on the population density of the phyllophagan. The intraspecific structuredness of the population in relation to the size of females and its transformation by the moment of their emergence promotes the evolutionary stability of the degree of fecundity of the sawfly.     

Johnston's organ in Neodiprion sertifer (Insecta: Hymenoptera)

The fine structure of Johnston's organ in the pine sawfly, Neodiprion sertifer, was studied by electron microscopy to determine if there exists a dimorphism in this organ corresponding to the sexual dimorphism in antennal shape and surface area. The organ is made up of scolopidia that are ultrastructurally similar to those of other insects. The scolopidia, identical in both sexes, comprise three sensory cells bearing two types of sensory processes: Two are shorter and smaller in diameter than the third, which extends into the cuticle of the membrane connecting pedicel and flagellum and terminates at an epicuticular invagination. The dendrites and sensory processes are surrounded by two types of enveloping (glial) cells-a scolopale cell and an attachment cell. Other enveloping cells occur at different levels of the scolopidium. Sexual dimorphism is evident only in the numbers of scolopidial groups: Males have more groups with fewer scolopidia, but both sexes possess about the same total number of scolopidia.     

Transmission agents of the nuclear polyhedrosis virus ofNeodiprion sertifer [Hym.: Diprionidae]

Colonies ofNeodiprion sertifer (Geoffroy) larvae infected by nuclear polyhedrosis virus were visited by parasitoids, predators, and scavengers. Six transmission pathways were studied in detail. Viable virus polyhedra (PIB) were isolated from external surfaces of parasitic wasps and faeces of tachinid flies, coccinellid larvae, and scavenging flies. Faeces of the scavenging flies remained infectious 9 months on foliage in the field. A bird fed on earthworms from contaminated soil passed infectious faeces. Raindrops on foliage below diseased larval colonies contained more than 10⁸ PIB/ml. It was concluded that although there are numerous transmission pathways, only a few may be available at any one time. The stochastic nature of many pathways implies that the overall transmission rate is highly variable. These results are consistent with frequent observations of low transmission levels between larvae of the same generation.      

A test of the sympatric host race formation hypothesis in Neodiprion (Hymenoptera: Diprionidae)

Theory suggests that sympatric speciation is possible; however, its prevalence in nature remains unknown. Because Neodiprion sawflies are host specialists and mate on their hosts, sympatric speciation via host shifts may be common in this genus. Here, we test this hypothesis using near-complete taxonomic sampling of a species group, comprehensive geographical and ecological data, and multiple comparative methods. Host-use data suggest that host shifts contributed to the evolution of reproductive isolation in Neodiprion and previous work has shown that gene flow accompanied divergence. However, geographical data provide surprisingly little support for the hypothesis that host shifts occurred in sympatry. While these data do not rule out sympatric host race formation in Neodiprion, they suggest that this speciation mode is uncommon in the genus and possibly in nature.     

Virulence of a nucleopolyhedrovirus to Neodiprion abietis (Hymenoptera: Diprionidae)

Different larval stages of balsam fir sawfly, Neodiprion abietis (Harris) (Hymenoptera: Diprionidae), were challenged by different concentrations of a nucleopolyhedrovirus in the laboratory to determine larval susceptibility to the virus and to test treatment effects of the virus on the sawfly survivors. The results indicated that younger larvae were more susceptible to the virus than older instars. The speed of larval death depended on the larval age and the virus concentration. Generally, the virus killed second or third instars in approximately 5 d and fourth or fifth instars in 10-12 d at concentrations of 10(7) polyhedral inclusion bodies (PIB) /ml. The virus had profound treatment effects on sawfly survivors. Feeding activity of the survivors was reduced by > 40% compared with that of the control group, pupal weight by approximately 25%, and adult emergence by > 30%. There was also a higher percentage of male adults in the virus-treated groups than in the control.     

Phylogenetic analysis of nuclear and mitochondrial genes reveals evolutionary relationships and mitochondrial introgression in the sertifer species group of the genus Neodiprion (Hymenoptera: Diprionidae)

Neodiprion Rohwer (Hymenoptera: Diprionidae) is a Holarctic genus of conifer-feeding sawflies with a remarkable amount of inter- and intraspecific diversity in host use, behavior, and development. This variation is thought to play a central role in Neodiprion diversification, but speciation hypotheses remain untested due to a lack of a robust phylogenetic estimate. Here, we utilize sequence data from three nuclear genes (CAD, ANL43, EF1alpha) to obtain a phylogenetic estimate for the genus. These analyses suggest that: (1) North American and Eurasian Neodiprion are monophyletic sister clades, (2) the sertifer group is paraphyletic with respect to the monophyletic lecontei group, and (3) on at least two occasions, dispersal from eastern to western North America proceeded via southern host bridges. Based on these results and host biogeography, we revise a previous scenario for the evolution of Neodiprion and suggest maximum ages for the genus and for the lecontei group (25 My and 14 My, respectively). In addition, because a previous study reported rampant mitochondrial introgression in the lecontei group, we assess its prevalence in the sertifer group. Analysis of three mitochondrial genes (COI, tRNA-leucine, and COII) reveals that mito-nuclear discordance is prevalent in the sertifer group, and patterns of species monophyly are consistent with those expected under frequent mitochondrial introgression. As was the case for lecontei group species, we find that introgression appears to be most pronounced between species that occasionally share hosts, suggesting that divergent host use is an important barrier to gene flow in Neodiprion. Finally, we suggest that the lack of phylogenetic resolution and prevalence of species non-monophyly in the non-Pinus feeding Neodiprion may result from the rapid divergence (possibly with gene flow) of these species following their entry into a novel adaptive zone.     

Zur Kenntnis der Kleinen Kiefern-Buschhornblattwespe,Diprion (Microdiprion) pallipes (Fall.) (Hym., Diprionidae)

Contributions to the knowledge of Diprion (Microdiprion) pallipes (Fall.) (Hym., Diprionidae). 1. Systematics, distribution and morphology Studies were made on D. pallipes var. politum injurious to young pine stands near Munich as well in laboratory as in high-fens and pine cultures. The Tenthredinid is native within the Alps and their foreland and had migrated northward along the river Isar in last decades. Part 1 deals with problems of systematics, distribution and morphology. D. pallipes (Fall.) is to be found in two European areas: a nominate race (D. pallipes pallipes) in northern Europe and a southern race (D. pallipes politum) in the alpine region. The original food plant of the alpine race is Pinus mugo Turra. The most important morphological characteristics of the developmental stages of the subspecies are described.