Metabolism and excretion of acetylchromenes by the migratory grasshopper

The extent of metabolism and excretion of three acetylchromenes (two toxic, one relatively nontoxic) were examined in adult migratory grasshoppers (Melanoplus sanguinipes) following topical administration. Both the total amount excreted (parent plus metabolites) and the proportion of parent compound in the excreta were inversely correlated with contact toxicity. Both toxic and nontoxic acetylchromenes are rapidly absorbed from the cuticle, with maximum excretion of parent and metabolite chromenes from 4 to 8 h posttreatment in each case. Much of the applied compounds (60–80%) apparently remains within the insect, and cannot be recovered by extraction of the insect. Metabolites formed result from simple oxidative and reductive transformations. For all of the compounds tested (including the allatocidin precocene II), the major mode of metabolism results from aliphatic hydroxylation of one of the geminal methyl groups on the chromene. No conjugated metabolites were found in the excreta.     

Precocene causes male determination in the aphid Myzus persicae

When adult apterous viviparous females of Myzus persicae, reared in short night conditions at 21–23°C, are treated with the precocene analogue 6-methoxy-7-ethoxy-2,2-dimethylchromene, they deposit males towards the end of their reproductive lives. The first-born normal daughters of the treated females also deposit some males at various times in their reproductive lives. Karyotypic analysis was used to investigate the sequence of male and female embryos in the ovarioles of precociously metamorphosed aphids. The experiments support the hypothesis (Mittler et al., 1979) that juvenile hormone level controls the sex determination process in aphids. Since male aphids have an XO sex chromosome constitution, this implies that juvenile hormone level influences the behaviour of the X-chromosomes at or before the single maturation division of the egg. At this division one X-chromosome is eliminated from eggs which will develop as males. Aphids provide the first example of a specific endocrine influence on chromosome behaviour in sex determination.     

Involvement of an Inducible Cytochrome P-450 in Precocene II Oxidation by Streptomyces Griseus

Streptomyces griseus oxidizes the insecticide precocene II to its cis- and trans-dihydrodiols and 3-chromenol after growth on an enriched medium containing soybean flour. Oxidation of precocene II is dependent on the level of cytochrome P-450 in this organism. Extracts of cells grown on media lacking soybean flour were devoid of cytochrome P-450 and could not oxidize precocene II. In an in vitro reconstituted system containing NADPH, spinach ferredoxin reductase, spinach ferredoxin and ammonium sulfate fractions enriched in cytochrome P-450, precocene II was oxidized to its dihydrodiols. An aerial mycelium-negative variant of S. griseus (AMY mutant), that was unable to elicit cytochrome P-450 when grown on soybean flour-enriched medium, failed to oxidize precocene II.     

Precocious metamorphosis of the aphid Myzus persicae induced by the precocene analogue 6-methoxy-7-ethoxy-2,2-dimethylchromene

For up to three days after being treated with the precocene analogue 6-methoxy-7-ethoxy-2,2-dimethylchromene, adult green peach aphids (Myzus persicae) gave birth to offspring (alate and apterous virginoparae, males) which underwent precocious metamorphosis. Only a few precocious gynoparae and no precocious oviparae were recorded. The precocious aphids became adultoid in the third or fourth-instar as a result of corpus allatum destruction. They were able to develop mature embryos, but could not deposit them because of incomplete differentiation of the reproductive tract. The treatment did not induce the production of alate virginoparae in the experimental clone of Myzus persicae. However, a few males were born late in the reproductive sequence of treated apterae. The corpora allata of the treated adults appeared on histological examination to be unaffected by precocene.     

Role of Corpus Allatum on the Modulation of Feeding Rhythm in the Semilooper Caterpillar,Achaea Janata (L)

A circadian feeding rhythm which may be entrained by photoperiod was found in fifth instarcaterpillars of the lepidopteran, Achaea Janata (L.). AUatectomy reduced the amount of food consumed; this consumption was significantly lower during the fifth instar in both allatectomized and sham-operated insects. An apparent circadian feeding pattern appeared on day 2 in the sham-operated caterpillars. Topical application of the anti-allatin, Precocene-II (50μg/animal) also reduced leaf consumption significantly compared to the respective controls, although these controls maintained the same apparent circadian feeding rhythm on day 2.     

Hormonal Control of Egg Dumping and Guarding in the Lace Bug, Gargaphia solani (Hemiptera: Tingidae)

This study examines the effect of juvenile hormone (JH) or a similarly acting juvenoid on the expression of maternal egg guarding and its life history alternative, egg dumping, in the lace bug, Gargaphia solani Heidemann. JH manipulations were indirect: methoprene, a synthetic JH analog, and precocene II, an allatocidal phytochemical commonly used to reduce JH synthesis, were applied exogenously to test the hypothesis that high JH titers promote egg production and egg dumping behavior, while low titers terminate egg production and initiate maternal care. As predicted, egg dumpers treated with precocene II ignored dumping opportunities and became egg guarders. Similarly, egg guarders that were treated with methoprene became gravid within 2 days and abandoned their eggs to become egg dumpers. These manipulations suggest that hormones can trigger the expression of both egg dumping and egg guarding in G. solani even when environmental conditions are inappropriate.     

Detoxification of naturally occurring chromenes in larvae of the generalist herbivore Spodoptera littoralis (Noctuidae)

Five naturally occurring chromenes from the Asteraceae including the insecticidal compounds precocene II (1) and encecalin (2) were administered to last instar larvae of Spodoptera littoralis via the food or by topical contact. Metabolites formed and excreted via the frass were analysed by GC-MS and by direct comparison with reference compounds obtained by partial synthesis. In total 28 different metabolites were identified, many of them reported here for the first time. All metabolites detected originated from phase I reactions (most probably catalysed by Cytochrom P-450-dependent monooxygenases) by hydroxylation, demethylation or reduction of the parent chromenes. The resulting metabolites can be regarded as detoxification products based on previous structure-activity studies. The increased polarity of the metabolites will furthermore facilitate their excretion by the larvae compared to the more apolar parent chromenes. The largest number of metabolites (eight for each compound) was detected following oral treatment with precocene II and encecalin respectively. 3-Monool as well as 3,4-trans-diol derivatives predominated in the frass of larvae treated with the latter compounds whereas the 6-hydroxyethyl derivatives were the major metabolites of the other chromenes investigated. The patterns of metabolites originating from precocene II or encecalin were the same following oral application or topical treatment.     

Vitellogenin titre in haemolymph of Locusta migratoria in normal adults,after ovariectomy,and in response to methoprene

Vitellogenin in the haemolymph of Locusta migratoria was assayed by rocket immunoelectrophoresis to elucidate aspects of its regulation. In many normal adult females, vitellogenin first appeared on days 5–9, rose quickly to peak levels, and declined before a second vitellogenic cycle; in others, it appeared later and built up more slowly. The timing of first appearance of vitellogenin, and proportions of early and late-developing individuals, differed markedly in groups from the same colony assayed in different years, suggesting effects of both genetic and environmental variation. Average peak levels of vitellogenin were 25–30 mg/ml. After ovariectomy, vitellogenin appeared near the normal time and increased for several weeks to about 300 mg/ml; haemolymph volume also increased greatly, so that the total haemolymph-vitellogenin pool reached about 300 mg/individual, or 100 times the normal amount. After ovariectomy, no cyclicity of vitellogenin accumulation was apparent. These results show that the ovary is not required for stimulation of vitellogenin synthesis, and suggest that normal cycling may depend on inhibition by the mature ovary. Females treated with ethoxyprecocene on day 1 of adult life to inactivate the corpora allata did not produce vitellogenin, but were induced to do so with the juvenile hormone analogue, methoprene. After injection of 150 μg of methoprene in mineral oil, there was one day lag, then vitellogenin increased in the haemolymph to the normal peak level and declined slowly to zero during 5 weeks; after a second injection of methoprene, vitellogenin re-appeared more rapidly, with less lag, reflecting accelerated secondary hormonal stimulation of vitellogenin synthesis in the fat body. Adult males showed no detectable haemolymph vitellogenin even after injection of large doses of methoprene.     

Behavioral changes and growth inhibition in last instar larvae of Heliothis zea induced by oral and topical application of precocene II

In diet choice tests, newly-ecdysed fifth instar Heliothis zea larvae randomly chose diet containing 4 mM precocene II or diet without precocene II but initially preferred to feed on diet without precocene II. After two days, however, they were found more frequently on diet containing precocene II. Nevertheless, larval growth and development were inhibited when forced to eat diet containing precocene II. Removal of the maxillae, the site of the primary gustatory receptors, did not prevent the growth inhibition caused by larval consumption of diet with precocene II. Topical treatments with precocene II also caused larval growth inhibition, and daily treatments were more detrimental to the larvae than a large single dose. Addition of methylenedioxyprecocene (6,7-methylenedioxy-2,2-dimethylchromene) to the diet did not reverse the observed larval growth inhibition and this compound was itself significantly toxic. These results indicate that larval growth and development are disrupted after precocene II ingestion, but not because of preingestive discrimination by olfaction and gustation.     

Antigonadotropic and antiallatal effects of precocene II in the firebrat, Thermobia domestica (Thysanura,Lepismatidae)

The sensitivity of an apterygote insect to precocene II was investigated for the first time in females of Thermobia domestica. Topical applications of this compound, which has been said to possess anti-juvenile hormone activity, were carried out at various times in the postecdysial period of a reproductive cycle, during intense vitellogenesis in the terminal oöcytes. The effects of treatment were observed on mortality, fecundity, insemination and also on the ultrastructure of corpora allata; they depend on the dose used and on the day of treatment. A single application of 10 μg/insect at the beginning of the postecdysial period induces antigonadotropic effects in females reared both with and without males. The effects of precocene add to those of non-insemination, which also elicits inhibition of oöcyte maturation. In addition, the cytotoxic action of precocene, leading to degeneration of the corpora allata, which is most probably irreversible, was demonstrated.