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Injection of the hormone 20-hydroxyecdysone (20-E) into partially fed (virgin) female adults of the American dog tick, Dermacentor variabilis, while they are attached and feeding on the rabbit host, initiated the expression of the vitellogenin (Vg) gene, and Vg protein secretion and uptake by the ovary. The induction of egg production by 20-E in this bioassay was dose dependent in the range of 1-50 times the concentration normally found in a replete, vitellogenic female. Ticks examined 4 d after the 50 x treatment were still attached to the host, had numerous enlarged vitellin-filled (brown) oocytes in their ovaries, but had not engorged to repletion. The ovaries reached weights similar to those found in untreated, replete (mated) females (pre-oviposition) while solvent-injected controls demonstrated no increase in oocyte size or increase in ovary weight. An increase in the levels of a putative Vg protein was observed in hemolymph samples collected 1, 2 and 3d post-20-E injection but was not observed in the corresponding solvent controls as determined by native PAGE. Analysis of the ecdysteroid-induced protein by tryptic digestion-mass fingerprinting and BLASTP found that the putative Vg had the strongest match to GP80 (U49934), the partial sequence for the vitellogenin protein from Boophilus microplus. A partial Vg cDNA was cloned and sequenced from replete females of D. variabilis with a high similarity to GP80. Using this message as a probe, Northern blots conducted with RNA collected from partially fed, virgin females 1, 2 and 3d post-20-E injection showed upregulation of the Vg mRNA on all 3 days. Controls injected with solvent only showed no Vg mRNA. Injections with juvenile hormone III did not stimulate Vg expression, oocyte growth or full engorgement. These studies indicate that ecdysteroids and not JH can initiate expression of the Vg gene, Vg protein synthesis and release into hemolymph, and Vg uptake into developing oocytes under bioassay conditions mimicking normal feeding on the host.

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Ionizing radiation is used as a phytosanitary treatment to mitigate risks from invasive species associated with trade of fresh fruits and vegetables. Commodity producers prefer to irradiate fresh product stored in modified atmosphere packaging that increases shelf life and delays ripening. However, irradiating insects in low oxygen may increase radiation tolerance, and regulatory agencies are concerned modified atmosphere packaging will decrease efficacy of radiation doses. Here, we examined how irradiation in a series of oxygen conditions (0.1-20.9 kPa O2) alters radiotolerance of larvae and pupae of a model lepidopteran Trichoplusia ni (Hubner) (Diptera: Noctuidae). Irradiating in severe hypoxia (0.1 kPa O2) increased radiation tolerance of insects compared with irradiating in atmospheric oxygen (20.9 kPa O2). Our data show irradiating pharate adult pupae at 600 Gy in moderately severe hypoxia (5 kPa O2) increased adult emergence compared with irradiation in atmospheric oxygen (20.9 kPa O2). Our data also show that in one of the three temporal replicates, irradiating T. ni larvae in moderately severe hypoxia (5 kPa O2) can also increase radiotolerance at an intermediate radiation dose of 100 Gy compared with irradiating in atmospheric oxygen conditions, but not at higher or lower doses. We discuss implications of our results in this model insect for the current generic doses for phytosanitary irradiation, including the recently proposed 250 Gy generic dose for lepidioptera larvae, and temporary restriction on irradiating commodities in modified atmosphere packaging that reduces the atmosphere to < 18 kPa O2.

BACKGROUND: Phytosanitary irradiation is a sustainable alternative to chemical fumigants for disinfesting fresh commodities from insect pests. However, irradiating insects in modified atmospheres with very low oxygen (<1 kPa O 2 ) has repeatedly been shown to increase radioprotective response. Thus, there is a concern that modified atmosphere packaging could reduce the efficacy of phytosanitary irradiation. One hurdle slowing the widespread application of phytosanitary irradiation is a lack of knowledge about how moderate levels of hypoxia relevant to the modified atmosphere packaging of most fresh commodities (3-10 kPa O 2 ) may affect phytosanitary irradiation treatments. Therefore, we hypothesize that critical PO 2 (P crit ), the level of oxygen at which an insect's metabolism becomes impaired, can be used as a diagnostic biomarker to predict the induction of a radioprotective response.RESULTS: Using the cabbage looper Trichoplusia ni (Hübner), we show that there is a substantial increase in radiation resistance when larvae are irradiated in atmospheres more hypoxic than their P crit (3.3 kPa O 2 ). These data are consistent with our hypothesis that P crit could be used as a diagnostic biomarker for what levels of hypoxia may induce radioprotective effects that could impact phytosanitary irradiation treatments.CONCLUSION: We propose that the relationship between P crit and radioprotective effects could allow us to build a framework for predicting the effects of low-oxygen atmospheres on the efficacy of phytosanitary irradiation. However, more widespread studies across pest species are still needed to test the generality of this idea.

This research suggests that aliphatic PEG polymers can be used as a new method for increasing the activity of insecticidal proteins.