SummaryThe green-fluorescent protein (GFP) from Aequorea victoria has been shown to be a convenient and flexible reporter molecule within a variety of eukaryotic systems, including higher plants. It is particularly suited for applications in vivo, since the mechanism of fluorophore formation involves an intramolecular autoxidation and does not require exogenous co-factors. Unlike standard histochemical procedures of fixation and staining required for analysis of the cellular or tissue-specific expression of other popular reporter molecules, such as the I]-glucuronidase (GUS) marker, analysis of GFP can be done in living cells with no specific pretreatments. This implies that GFP might also be particularly suited for studies of intracellular protein targeting. In this paper, the use of GUS is compared with that of GFP for the analysis of nuclear targeting in tobacco. A novel oligopeptide motif from a tobacco protein is described which confers nuclear localization of GUS. The use of this oligopeptide and two from potyviral proteins to target GFP to the nucleus is examined. An essential modification of GFP is described, which specifically increases its molecular weight to eliminate its passive penetration into the nucleus. Three examples of the targeting of these enlarged GFP molecules to the nucleus are illustrated. GFP, in combination with confocal microscopy, offers significant advantages over traditional methods of studying nuclear targeting.

The proximate forces that create omnivores out of herbivores and predators have long fascinated ecologists, but the causal reasons for a shift to omnivory are poorly understood. Determining what factors influence changes in trophic position are essential as omnivory plays a central role in theoretical and applied ecology. We used sevenspotted lady beetles (Coccinella septempunctata) to test how prey nutrient content affects beetles’ propensity to engage in herbivory. We show that beetles consuming an all‐prey diet demonstrate normal growth and development, but suffer a complete loss of fitness (spermatogenic failure) that is restored via herbivory and supplementation with phytosterols and cholesterol. Furthermore, we show that lady beetles possess a state‐dependent sterol‐specific appetite and redressed their sterol deficit by feeding on foliage. These results demonstrate that predators balance their nutrient intake via herbivory when prey quality is low, and reveal a selective force (sterol nutrition) that drives predatory taxa to omnivory.

Plant produced insect molting hormones, termed phytoecdysteroids (PEs), are thought to function as plant defenses against insects by acting as either feeding deterrents or through developmental disruption. In spinach (Spinacia oleracea), 20-hydroxyecdysone (20E) concentrations in the roots rapidly increase following root damage, root herbivory, or methyl jasmonate (MJ) applications. In this inducible system, we investigated the plant defense hypothesis by examining interactions of roots, 20E concentrations, and larvae of the dark-winged fungus gnat (Bradysia impatiens). Root herbivory by B. impatiens larvae resulted in a 4.0- to 6.6-fold increase in root 20E concentrations. In paired-choice tests, increases in dietary 20E stimulated B. impatiens feeding deterrency. B. impatiens larvae preferred control diets, low in 20E, to those constructed from induced roots and those amended with 20E (25 to 50 micro g/g wet mass). When confined to 20E-treated diets, concentrations as low as 5 micro g/g (wet mass) resulted in significantly reduced B. impatiens survivorship compared to controls. The induction of root 20E levels with MJ resulted in a 2.1-fold increase in 20E levels and a 50% reduction in B. impatiens larval establishment. In a paired-choice arena, untreated control roots were damaged significantly more by B. impatiens larvae than MJ-induced roots that contained 3-fold greater 20E levels. Based on dietary preference tests, the 20E concentrations present in the MJ-induced roots (28 micro g/g wet mass) were sufficient to explain this reduction in herbivory. Interactions between spinach roots and B. impatiens larvae demonstrate that PEs can act as inducible defenses and provide protection against insect herbivory.

Summary1. Arthropods, including insects, are unique among animals in that they cannot synthesize sterols, including cholesterol, de novo. Some phytophagous insects (e.g. caterpillars, grasshoppers) generate tissue cholesterol by metabolizing plant phytosterols. Currently, little is known about sterols in plant phloem sap, and their significance to the nutritional physiology and ecology of phloem-feeding insects.2. The sterol profiles of leaves from two plant species, Chinese cabbage and tobacco, were dominated by the phytosterols sitosterol and stigmasterol, respectively. In contrast, the principle sterol in the phloem sap of both plants was cholesterol, which is traditionally considered an animal sterol. Cholesterol was also the most abundant sterol in the carcass and honeydew of Myzus persicae aphids feeding on these plants. 3. The effect of sterol structure on M. persicae was investigated using modified tobacco plants that contained high levels of atypical steroids, specifically ketone-steroids. Aphids reared on the modified tobacco plants had a high atypical steroid content, severely reduced reproduction and high mortality. 4. Our data indicate that the total sterol composition of plants is not necessarily representative of the sterol profile available to phloem-feeding insects, and that the sterol utilization patterns of phloem-feeding insects may differ from chewing insect herbivores utilizing the same plant. Atypical steroids are naturally at insufficient concentrations for significant deleterious effects on insect herbivores, and possible reasons why plants apparently do not use them as defensive compounds are considered.

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