Sawfly Athalia Rosae Research Articles

Survival of the Sawfly Athalia rosae Upon Infection by an Entomopathogenic Fungus and in Relation to Clerodanoid Uptake.

Larvae of the turnip sawfly Athalia rosae are a pest of Brassicacae plants, as their feeding can cause defoliation of various crops of economic importance. The larvae and the adults of this sawfly species are known to take up different classes of chemical compounds from their respective host plants, with potentially deterrent functions against predators. In addition, compounds taken up by the adults, the clerodanoids, are known for their antimicrobial activity. These features could be a challenge to biocontrol strategies. Several natural enemies of A. rosae have been identified, targeting larval and pupal stages of A. rosae, which could potentially be used as biocontrol agents. However, targeting the adult stage of a larval pest in addition to targeting the juvenile stages may improve population control. In this study, we ask whether a strain of the entomopathogenic fungus Beauveria bassiana shows biological activity against A. rosae adults. We also investigate whether the behavior of clerodanoid uptake by the adults, which is commonly found, affects their survival in response to a B. bassiana exposure. We found a clear dose-response relationship, i.e., with increasing fungal conidia concentrations survival of A. rosae decreased. However, there was only a low incidence of mycelial growth and sporulation from A. rosae cadavers, indicating that either the fungus is not successfully developing inside this host, or it is not able to re-emerge from it. Clerodanoid uptake decreased the survival of healthy adults; however, it did not increase their survival to B. bassiana. Our results revealed that this strain of B. bassiana if applied alone is probably not suitable for biocontrol of this sawfly species, because A. rosae showed a high baseline resistance against this fungus. The behavior of clerodanoid uptake is unlikely to have evolved as a defense against this entomopathogenic fungus.

Wing vein development in the sawfly Athalia rosae is regulated by spatial transcription of Dpp/BMP signaling components

Wing venation among insects serves as an excellent model to address how diversified patterns are produced. Previous studies suggest that evolutionarily conserved Decapentaplegic (Dpp)/Bone Morphogenetic Protein (BMP) signal plays a critical role in wing vein development in the dipteran Drosophila melanogaster and the hymenopteran sawfly Athalia rosae. In sawfly, dpp is ubiquitously expressed in the wing during prepupal stages, but Dpp/BMP signal is localized in the future vein cells. Since localized BMP signaling involves BMP binding protein Crossveinless (Cv), redistribution of BMP ligands appears to be crucial for sawfly wing vein formation. However, how ubiquitously expressed ligands lead to a localized signal remains to be addressed. Here, we found that BMP binding protein short gastrulation (Sog) is highly expressed in the intervein cells. Our data also reveal that BMP type I receptors thickveins (Tkv) and saxophone (Sax) are highly expressed in intervein cells and at lower levels in the vein progenitor cells. RNAi knockdown of Ar-tkv or Ar-sax indicates that both receptors are required for localized BMP signaling in the wing vein progenitor cells. Taken together, our data suggest that spatial transcription of core- and co-factors of the BMP pathway sustain localized BMP signaling during sawfly wing vein development.

Wolbachia infections of the butterfly Eurema mandarina interfere with embryonic development of the sawfly Athalia rosae

Although maternally transmitted microorganisms such as Wolbachia are well known to have a variety of effects on the reproduction of diverse insect species, little is known about the underlying mechanisms of actions. Artificial transfer of Wolbachia between taxonomically distant host species may provide insights into Wolbachia-induced manipulations of hosts. Here we performed a cross-order transfer of feminizing Wolbachia derived from a butterfly, Eurema mandarina. The Wolbachia were propagated in the Eurema hecabe cell line, called NTU-YB, and then used to inject prepupal/pupal females of a Wolbachia-free hymenopteran sawfly, Athalia rosae. The 14 females that emerged as adults looked morphologically and behaviorally healthy, and ovarian development appeared normal on dissection. However, in contrast to the control, none of the 333 eggs harbored by the seven Wolbachia-injected females developed successfully. Similarly, none of the 140 eggs laid on host plant by the four Wolbachia-injected females, which were mated with males, showed any signs of development. Wolbachia infection was detected from whole-body samples of the inoculated individuals, but not from the eggs they produced. Disruption of embryonic development despite the absence of Wolbachia in the egg cytoplasm may represent a new phenotype involving maternal effects that result in female sterility.

Uncovering different parameters influencing florivory in a specialist herbivore

1. It has become increasingly recognised that several herbivores switch from folivory (leaf-feeding) to florivory (flower-feeding) during larval development. Yet, it remains poorly understood which cues influence this behaviour, whether a switch to florivory is consistently shown on different hosts, and to what extent florivory could be hindered by plant traits. 2. Using the sawfly Athalia rosae and two Brassicaceae differing in architecture and surface structure, the cues that influence larval movement to the flowers were investigated. A broad set of behavioural assays was employed and physical and chemical plant traits potentially affecting the larvae were analysed. Furthermore, the consequences of folivory versus florivory on insect performance were studied. 3. The larvae preferred flowers over leaves. Consumption of particular flower parts correlated partly with measured plant traits such as glucosinolate distribution. Visual cues were of higher importance than volatile cues. The initial position of newly hatched larvae on plants influenced the probability of the larvae reaching the flowers during development. Trichomes and surface waxes hindered the larvae from moving upwards to the flowers. Larvae developed slower and gained less body mass when feeding on inflorescences of Brassica nigra than when feeding on leaves, in contrast to the patterns observed before on Sinapis alba, where florivory led to an improved performance. 4. This study demonstrates that florivory depends on various host plant traits. It reveals new insights into different parameters influencing this multifaceted phenomenon and into the expected impact on the ecology and fitness of both the attacking herbivores and the plants.

Insights into the molecular mechanisms underlying diversified wing venation among insects.

Insect wings are great resources for studying morphological diversities in nature as well as in fossil records. Among them, variation in wing venation is one of the most characteristic features of insect species. Venation is therefore, undeniably a key factor of species-specific functional traits of the wings; however, the mechanism underlying wing vein formation among insects largely remains unexplored. Our knowledge of the genetic basis of wing development is solely restricted to Drosophila melanogaster. A critical step in wing vein development in Drosophila is the activation of the decapentaplegic (Dpp)/bone morphogenetic protein (BMP) signalling pathway during pupal stages. A key mechanism is the directional transport of Dpp from the longitudinal veins into the posterior crossvein by BMP-binding proteins, resulting in redistribution of Dpp that reflects wing vein patterns. Recent works on the sawfly Athalia rosae, of the order Hymenoptera, also suggested that the Dpp transport system is required to specify fore- and hindwing vein patterns. Given that Dpp redistribution via transport is likely to be a key mechanism for establishing wing vein patterns, this raises the interesting possibility that distinct wing vein patterns are generated, based on where Dpp is transported. Experimental evidence in Drosophila suggests that the direction of Dpp transport is regulated by prepatterned positional information. These observations lead to the postulation that Dpp generates diversified insect wing vein patterns through species-specific positional information of its directional transport. Extension of these observations in some winged insects will provide further insights into the mechanisms underlying diversified wing venation among insects.

Rapid incorporation of glucosinolates as a strategy used by a herbivore to prevent activation by myrosinases

Various plants have a binary defence system that consists of a substrate and a glucosidase, which is activated upon tissue disruption thereby forming reactive hydrolysis products. Insects feeding on such plants have to overcome this binary defence system or prevent the activation. In this study, we investigated the strategy used by a herbivore to deal with such binary defence. We studied, how the larvae of the sawfly Athalia rosae (Hymenoptera: Tenthredinidae) circumvent the activation of glucosinolates by myrosinase enzymes, which are found in their Brassicaceae host plants. Myrosinase activities were low in the front part of the larval gut but activities increased over the gut passage. In contrast, the glucosinolates were only highly concentrated in the first gut part and were rapidly incorporated into the haemolymph before the food reached the second half of the gut. Thus, the uptake and concentration of glucosinolates, i.e., sequestration, must occur in the front part of the gut. Using Matrix Assisted Laser Desorption Ionization-Mass Spectrometry Imaging (MALDI-MSI), we could demonstrate that the incorporated glucosinolate sinalbin circulates in the haemolymph where it accumulates around the Malpighian tubules. This study highlights the pivotal role of the gut of an adapted herbivore as a regulatory functional organ to cope with plant toxins. MALDI-MSI turned out as a highly useful technique to visualise glucosinolates in a herbivore, which has to deal with plants exhibiting a binary defence system, and may be applied to follow the fate of plant metabolites in other insect species in the future.

Dpp/BMP transport mechanism is required for wing venation in the sawfly Athalia rosae

The pattern of wing venation varies considerably among different groups of insects and has been used as a means of species-specific identification. However, little is known about how wing venation is established and diversified among insects. The decapentaplegic (Dpp)/bone morphogenetic protein (BMP) signaling pathway plays a critical role in wing vein formation during the pupal stages in Drosophila melanogaster. A key mechanism is BMP transport from the longitudinal veins (LVs) to the posterior crossvein (PCV) by the BMP-binding proteins, short gastrulation (Sog) and twisted gastrulation2/crossveinless (Tsg2/Cv). To investigate whether the BMP transport mechanism is utilized to specify insect wing vein patterns in other than Drosophila, we used the sawfly Athalia rosae as a model, which has distinct venation patterns in the fore- and hindwings. Here, we show that Ar-dpp is ubiquitously expressed in both the fore- and hindwings, but is required for localized BMP signaling that reflects distinct wing vein patterns between the fore- and hindwings. By isolating Ar-tsg/cv in the sawfly, we found that Ar-Tsg/Cv is also required for BMP signaling in wing vein formation and retains the ability to transport Dpp. These data suggest that the BMP transport system is widely used to redistribute Dpp to specify wing venation and may be a basal mechanism underlying diversified wing vein patterns among insects.

Effects of Indole Glucosinolates on Performance and Sequestration by the Sawfly Athalia rosae and Consequences of Feeding on the Plant Defense System

In this study, the performance and sequestration specificity of the sawfly Athalia rosae L. (Hymenoptera: Tenthredinidae) reared on two wild Brassica oleracea L. (Brassicaceae) populations (KIM and WIN), which express indole glucosinolates (GSs) in very high concentrations, were investigated. The local and systemic plant responses to larval feeding also were analyzed. Indole GSs predominated in KIM plants, whereas aliphatic GSs prevailed in the WIN population, which had several times higher total GS concentrations than KIM plants. Plants of the KIM population had lower specific leaf areas, and higher carbon/nitrogen ratios than WIN plants. The insects reared on WIN plants performed better for most traits than insects reared on the KIM population. The larvae preferentially sequestered aliphatic GSs but when feeding on KIM plants, indole GSs were also concentrated in the hemolymph. In response to feeding by A. rosae larvae, indole GSs were induced locally and systemically, whereas aliphatic GSs were reduced only in systemic leaves of the WIN population. Soluble myrosinase activities were constitutively higher in WIN than in KIM plants, and feeding damage led to a significant reduction of this glucosinolate-degrading enzyme in WIN plants only. The data suggest that the sawfly is well adapted to high concentrations of aliphatic GSs, which are sequestered by the larvae. In contrast, the larvae may be less adapted to plants dominated by indole GSs. Selective induction of indole GSs by the plants in response to larval feeding may be adaptive as accumulation of indole GSs may lower the performance of the larvae.

Matching plant defence syndromes with performance and preference of a specialist herbivore

Summary 1. Plants use various strategies to cope with opponents. According to the plant defence theory these traits are predicted to covary across taxa and were shown to be grouped into several syndromes for Apocynaceae. Specialist herbivores tolerate or detoxify components of the plants' chemical weapons. Their development might mirror the putative defence syndromes of their hosts. This hypothesis was tested by measuring nutritive values and potential defence properties of seven species of Brassicaceae, considering leaf age. Effects of these traits were assessed on various lifehistory traits of the oligophagous sawfly Athalia rosae. 2. Positive correlations were found between particular plant traits. A hierarchical cluster analysis assembled plants in three distinct groups with either low nutritional quality or higher nutritional quality together with either only chemical or with chemical and mechanical defences. Although young and old leaves of each species grouped within the same clusters, age was a significant source of variation, demonstrating that ontogenetic changes in plant quality influence associations. 3. The correlations of several life-history parameters of A. rosae with each other and with plant traits were investigated. Mortality rates, developmental times and adult mass were correlated and important for insect fitness. Preference of adult females was largely influenced by larval performance. Three distinct clusters were determined, with fitness of this specialist highly depending on host plant quality. 4. Multivariate studies of the long-term performance and preference of the specialist in relation to the plant defence syndromes revealed general implications for plant - insect interactions, namely that insect traits mirror the defence syndromes of their hosts. Larval performance and adult preference were more influenced by general mechanical and chemical than by chemical plant defence traits, to which this specialist is adapted. 5. The 'plant defence syndromes hypothesis' is of general importance, however, as defence strategies and nutritional value change drastically throughout ontogeny, tissue age should be considered, and modifications on single trait associations are needed. Clusters forming plant syndromes mostly matched with clusters determined from the parameters of a specialist herbivore. Further research is needed on generalist performance, which might be differently influenced by the plants' defence strategies.

Interactions between glucosinolate- and myrosinase-containing plants and the sawfly Athalia rosae

Several insects have specialised on using Brassicaceae as host plants. Therefore, they evolved metabolic pathways to cope with the defensive glucosinolate–myrosinase system of their diet. Larvae of the turnip sawfly, Athalia rosae L. (Hymenoptera: Tenthredinidae), incorporate various glucosinolates from their hosts into their haemolymph. The ability to sequester these metabolites makes A. rosae a useful model system to study mechanisms of glucosinolate metabolism in this species compared to other specialists, and to study effects of sawfly feeding on levels of glucosinolates and their hydrolysing enzymes in plants. The levels of plant metabolites might in turn directly affect the performance of the insect. On the one hand, costs for glucosinolate uptake and avoidance of myrosinase activity were postulated. On the other hand, sequestration of glucosinolates can be part of the insect’s defence against several predators. Here, the findings on glucosinolate metabolic pathways are compared between different herbivores and the sawfly. The impact of different glucosinolate levels and myrosinase activities on the performance of A. rosae is discussed. Furthermore, effects of feeding by A. rosae larvae on the chemical composition and enzyme activities of various Brassicaceae species are summarised. Induction patterns vary not only between different plant species and cultivars but also due to the inducing agent. Finally, the plant–herbivore interactions are discussed with regard to the sawflies’ defence abilities against different carnivore guilds.

Uptake and turn-over of glucosinolates sequestered in the sawfly Athalia rosae

Larvae of the sawfly Athalia rosae sequester glucosinolates from their various host plants of the Brassicaceae into their hemolymph for defensive purposes. We found that the glucosinolate concentration in the insect varies in a fluctuating manner during larval development. Analyses of larvae which had been offered diets with different glucosinolate profiles showed that there is an equilibrium between a rapid uptake of glucosinolates into the hemolymph and a continuous turn-over. Injection of glucotropaeolin into the hemolymph and ingestion of the same amount resulted in similar levels of intact glucosinolates recovered from larvae after different periods of time. This indicates that hemolymph glucosinolates are the principal source for glucosinolate degradation. Feeding experiments with [ 14C]-labeled glucotropaeolin revealed that the majority of the ingested glucosinolate is excreted as one or more unidentified metabolite(s) within 14 h. We found no indication for the presence of an insect myrosinase, or sulfatase in A. rosae, which have been shown to be involved in glucosinolate metabolism in other specialists feeding on Brassicaceae. Furthermore, the metabolism of sinalbin in A. rosae seems to result in different products than its metabolism in the caterpillar Pieris rapae. Obviously, A. rosae has yet another way of coping with the glucosinolates.

Host plant derived feeding deterrence towards ants in the turnip sawfly Athalia rosae

AbstractLarvae of the sawfly Athalia rosae ruficornis Jakovlev (Hymenoptera: Tenthredinidae) feed on several glucosinolate‐containing plants and have been shown to sequester the main glucosinolates of different hosts, namely sinalbin (p‐hydroxybenzylglucosinolate) from Sinapis alba L., sinigrin (allylglucosinolate) from Brassica nigra (L.) Koch, and glucobarbarin ((S)‐2‐hydroxy‐2‐phenylethylglucosinolate) from Barbarea stricta Andrz. (Brassicaceae). These plant metabolites are stored in the haemolymph, which is readily released when larvae are attacked by predators. In a dual‐choice bioassay the bio‐activity of sawfly haemolymph collected from larvae reared on different host plants (S. alba, B. nigra, and B. stricta) was tested against the ant Myrmica rubra L. (Hymenoptera: Formicidae). The haemolymph had a stronger deterrence effect when the corresponding sawfly larvae were reared on S. alba than when reared on B. nigra and B. stricta. Haemolymph of caterpillars of Pieris rapae L. (Lepidoptera: Pieridae) that had fed on S. alba was not deterrent to the ants. No sinalbin could be detected in their haemolymph. The glucosinolates sinalbin and sinigrin, offered in a concentration comparable to that in the sawfly haemolymph, were deterrent to the ants, but not as strongly as the corresponding haemolymph samples. This suggests, that glucosinolates are not the only compounds involved in the chemical defence of A. rosae. However, the presence of sequestered glucosinolates is already a sufficient defence towards predators such as ants, and their effectiveness is modulated by the host plant chemistry.

Sequestration of host plant glucosinolates in the defensive hemolymph of the sawfly Athalia rosae.

Interactions between insects and glucosinolate-containing plant species have been investigated for a long time. Although the glucosinolate-myrosinase system is believed to act as a defense mechanism against generalist herbivores and fungi, several specialist insects use these secondary metabolites for host plant finding and acceptance and can handle them physiologically. However, sequestration of glucosinolates in specialist herbivores has been less well studied. Larvae of the tumip sawfly Athalia rosae feed on several glucosinolate-containing plant species. When larvae are disturbed by antagonists, they release one or more small droplets of hemolymph from their integument. This "reflex bleeding" is used as a defense mechanism. Specific glucosinolate analysis, by conversion to desulfoglucosinolates and analysis of these by high-performance liquid chromatography coupled to diode array UV spectroscopy and mass spectrometry, revealed that larvae incorporate and concentrate the plant's characteristic glucosinolates from their hosts. Extracts of larvae that were reared on Sinapis alba contained sinalbin, even when the larvae were first starved for 22 hr and, thus, had empty guts. Hemolymph was analyzed from larvae that were reared on either S. alba, Brassica nigra, or Barbarea stricta. Leaves were analyzed from the same plants the larvae had fed on. Sinalbin (from S. alba), sinigrin (B. nigra), or glucobarbarin and glucobrassicin (B. stricta) were present in leaves in concentrations less than 1 micromol/g fresh weight, while the same glucosinolates could be detected in the larvae's hemolymph in concentrations between 10 and 31 micromol/g fresh weight, except that glucobrassicin was present only as a trace. In larval feces, only trace amounts of glucosinolates (sinalbin and sinigrin) could be detected. The glucosinolates were likewise found in freshly emerged adults, showing that the sequestered phytochemicals were transferred through the pupal stage.

Production of haploid-haploid chimeras by sperm injection in the sawfly, Athalia rosae (Hymenoptera).

Mature unfertilized eggs (oocytes) dissected from the ovary of the sawfly Athalia rosae (Hymenoptera) begin parthenogenetic development if exposed to distilled water and produce haploid males. Injection of sperm into mature oocytes through the anterior pole resulted in karyogamy in a fraction of cases which developed as diploid females. No haploid-haploid chimeras due to independent participation of the injected sperm in development were produced. When sperm were injected through the posterior pole, however, fertilization never occurred but haploid-haploid chimeras were produced in a smaller fraction of cases. Both egg nucleus-derived and injected sperm-derived nuclei contributed in forming the germ cells of the chimeric males.

Experiments inducing prospective polar body nuclei to participate in embryogenesis of the sawfly Athalia rosae (Hymenoptera)

Mature eggs dissected from ovaries of unmated females of Athalia rosae (Hymenoptera: Tenthredinidae), if placed on a filter-paper soaked with distilled water, are activated and develop to haploid males. Occasionally, however, diploid females develop from these artificially activated eggs. Treatment of mature unfertilized eggs dissected from diploid females with ice-cold temperatures immediately before activation and with a high temperature (36° C) upon and immediately after activation resulted in the production of diploid males, diploid females, triploid females and gynandromorphs at high frequency. The same treatment of mature unfertilized eggs dissected from triploid females resulted in the production of only triploid survivors. These results, together with the results on the segregation of a marker mutation, yellow fatbody (yfb), appear to indicate that meiotic divisions were complete in the treated eggs, and that all four nuclei became potentially capable of participating in development with or without automictic fusion.

The Wings of Insects. Chapter III. (Continued)

Previous articleNext article FreeThe Wings of Insects. Chapter III. (Continued)J. H. Comstock, and J. G. NeedhamJ. H. Comstock Search for more articles by this author , and J. G. Needham Search for more articles by this author PDFPDF PLUS Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by The American Naturalist Volume 32, Number 377May, 1898 Published for The American Society of Naturalists Article DOIhttps://doi.org/10.1086/276868 Views: 110 Citations: 3Citations are reported from Crossref PDF download Crossref reports the following articles citing this article:Laura C.V. Breitkreuz, Shaun L. Winterton, Michael S. Engel Wing Tracheation in Chrysopidae and Other Neuropterida (Insecta): A Resolution of the Confusion about Vein Fusion, American Museum Novitates 3890, no.38903890 (Dec 2017): 1–44.https://doi.org/10.1206/3890.1Shinya Matsuda, Naotoshi Yoshiyama, Jaana Künnapuu-Vulli, Masatsugu Hatakeyama, Osamu Shimmi Dpp/BMP transport mechanism is required for wing venation in the sawfly Athalia rosae, Insect Biochemistry and Molecular Biology 43, no.55 (May 2013): 466–473.https://doi.org/10.1016/j.ibmb.2013.02.008Charles Robertson Homologies of the Wing Veins of Hymenoptera, Science 11, no.264264 (Jan 1900): 112–113.https://doi.org/10.1126/science.11.264.112