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.

Quantity and quality of venom released by a spider (Cupiennius salei, Ctenidae).

The amount of venom injected by the spider Cupiennius salei depended on the efficiency of the mechanical defence of the prey species. Spiders were milked for the first venom (i.e. the first microlitre of venom emitted) versus remaining venom, and for venom regenerated from emptied glands. HPLC gel filtration and IEF electrophoresis showed that the protein content of the first venom was only half as compared to that of the remaining venom, and that this was due to a dilution of all proteins. Venom regeneration came in two speeds. The amount of venom was regenerated more rapidly than the protein concentration. Newly regenerated venom as compared to older venom was characterized by a lower concentration of all proteins and by a higher total concentration of free amino acids, whereas histamine and taurine did not follow this trend. K+ concentration and pH remained similar during venom regeneration. Crickets injected with the venoms showed less acute symptoms when the protein concentration was lower, namely with the first venom and with newly regenerated venom. Consequently, a spider which modulates the quantity of venom injected into a prey also directly changes the venom quality. The ecological consequences of this are discussed. This paper also discusses which region of a gland (ampulla, extracellular and intracellular parts of the glandular sac) is involved in the changes of the venom quality.

Sequestration ofVeratrum alkaloids by specialistRhadinoceraea nodicornis konow (Hymenoptera, Tenthredinidae) and its ecoethological implications.

The larvae of the specialist sawflyRhadinoceraea nodicornis Konow (Hymenoptera, Tenthredinidae) store in their hemolymph ceveratrum alkaloids originating from the host plantVeratrum album L. (Liliales, Melanthiaceae). The major alkaloid found in the hemolymph is 3-acetyl-zygadenine. Qualitative and quantitative data showed that the plant alkaloid 3-angeloylzygadenine is most probably metabolized in the larval gut to zygadenine and then acetylated. A still unidentified alkaloid with a molecular weight of 591 Da was detected in plant leaves as well as in the gut, hemolymph, and excrement of larvae. Protoveratrine A and B, on the other hand, seem to be degraded by the larvae. These findings indicate that the pathway of ceveratrum alkaloids inR. nodicornis larvae is fourfold: direct sequestration, metabolism followed by sequestration, excretion of intact alkaloids, and degradation. In contrast, no ceveratrum alkaloids were detected in the hemolymph and excrement of larvae of the generalist sawflyAglaostigma sp. fed withV. album leaves. Bioassays with the antMyrmica rubra L. proved that the hemolymph ofR. nodicornis larvae is highly deterrent and toxic. In bioassays evaluating defensive efficiency against predators (ants, spiders, and bushcrickets), no larvae were eaten. Ceveratrum alkaloids were also detected in the hibernating prepupae ofR. nodicornis. In feeding bioassays, the shrewCrocidura russula Hermann rarely fed upon prepupae, suggesting that this stage is also protected from predation to some degree. In field surveys, the only parasitoids recorded were two ichneumonid species that are believed to be specialized onR. nodicornis. Bioassays and field observations enable us to suppose thatR. nodicornis and its enemies produce a food web of ion connectance.

Gregariousness, field distribution and defence in the sawfly larvae Croesus varus and C. septentrionalis (Hymenoptera, Tenthredinidae).

The larvae of Croesus varus are cryptic and those of C. septentrionalis aposematic. In both species, females lay eggs in groups, but the groups formed by young larvae are smaller in C. varus than in C. septentrionalis. In both species, group size decreases during successive larval instars, and the cryptic species becomes solitary, while the aposematic species remains gregarious during all larval instars. On Alnus shrubs in the field, the distribution of the groups ofC. septentrionalis larvae is more aggregative. They are found mainly on mediumsized shrubs (1.5-2.5 m high), whereas groups of C. varus larvae are also found on larger shrubs. Moreover, during the 2 years of observation, C. septentrionalis was present for a shorter time than C. varus, even when the former was much more abundant than the latter. Thus, conspicuousness of the aposematic species should be amplified by this spatial and temporal aggregation. The efficiency of both defensive strategies is compared and discussed, taking into account these particular field distributions, predation and some parasitism factors, and the chemical defences of both species.

Aldehydic contact poisons and alarm pheromone of the antCrematogaster scutellaris (Hymenoptera: Myrmicinae) : Enzyme-Mediated Production from acetate precursors.

The Dufour gland ofCrematogaster scutellaris stores a mixture of long-chain primary acetates bearing a cross-conjugated dienone (Scheme 1, la-c). The poison gland contains two highly active enzymes: an acetate esterase and an alcohol oxidase. During venom emission, the constituents of both glands mix and accumulate on the sting, where the formation of the highly electrophilic aldehydes (Scheme 1, 2a-c) from their acetate precursors is initiated. Acetic acid, produced during the reaction, acts as alarm pheromone. The toxicity of the acetates (Scheme 1, la-c) and of the crude secretion has been assessed by topical application onMyrmica rubra. The acetatecontaining secretion from the Dufour gland was less toxic than the enzymatically altered secretion that was rich in aldehydes. The production of acids (Scheme 1, 3a-c) was an artifact resulting from the nonenzymatic oxidation of the unstable aldehydes.

Modes of defense in nematine sawfly larvae : Efficiency against ants and birds.

Ventral glands are common in nematine larvae (Hymenoptera: Symphyta), but they show various degrees of development and are functional for defense only in some species. In those species, volatile irritants are produced which are effective against ants. Alternative or complementary mechanisms against ants are the pubescence ofTrichiocampus spp., the foam pillars constructed byStauronema compressicornis, various movements of the abdomen, which occur independently of the glandular secretion in several species, immobility of the flat larvae ofNematinus luteus, and burrowing within plant tissues in gallicolous larvae or miners. Glandular development is not clearly related to the appearance of the larvae, either cryptic or aposematic. The secretion, even when it is produced in large amounts by species with well-developed glands, is only moderately efficient against great tits. Bright colors are found in gregarious larvae; these were accepted only with reluctance by great tits and sometimes rejected, even species in which the ventral glands are reduced. We suggest that the various volatile irritants secreted by ventral glands are aimed primarily against insects (e.g., ants) and only secondarily against birds.