Temperature-allelochemical interactive effects on performance ofManduca sexta caterpillars from its host and certain nonhosts

A series of experiments was conducted to ascertain the effects of temperature and allelochemicals on the performance of an insect herbivore, with the goal of methodically expanding the cumulative data set on interactive effects of temperature and allelochemicals. The allelochemicals examined were caffeine, chlorogenic acid, quercetin, tannic acid and tomatine with the thermal regimes 20:15 °C versus 30:15 °C. Growth, molting time and food utilization efficiencies of third instar tobacco hornworms (Manduca sexta (L.)) were measured. Our results indicated that reductions in developmental rate by the phenolics were primarily due to effects occurring around and during molt initiation, that different phenolics may affect molt processes differently, and that some of the effects of the phenolics were a function of temperature, with greater negative effects at cool temperature. Negative effects of caffeine were most pronounced earlier in the stadium rather than during molt processes and, for some variables, the effect of caffeine was also a function of temperature.     

Effects of light availability on host plant chemistry and the consequences for behavior and growth of an insect herbivore

We examined how light availability influenced the defensive chemistry of tomato (Lycopersicon esculentum: Solanaceae). Tomato plants were grown either in full sunlight or under shade cloth rated at 73%. Leaves from plants grown in full sunlight were tougher, had higher concentrations of allelochemicals (chlorogenic acid, rutin and tomatine), and had less protein than leaves from plants grown in shade. We determined how these differences in host plant quality due to light availability affected the behavior and growth of a Solanaceae specialist, Manduca sexta. Both in the greenhouse and in the field, caterpillars on shade-grown plants grew heavier in a shorter amount of time than those on plants that had previously been grown in full sunlight. In contrast, the effects of previous light availability to plants on caterpillar behavior appeared to be minor.To further investigate how light availability to plants influenced herbivore growth, we examined the effects of leaf-powder diets made from tomato leaves of different ages (new, intermediate, or mature) grown in full sunlight or shade on caterpillar performance. Caterpillars fed diets made from plants grown in shade consumed less but grew faster than larvae fed diets made from tomato plants grown in full sunlight. Caterpillars fed diets made from new leaves grew larger in less time than caterpillars fed diets made from intermediate aged leaves. Caterpillars did not survive on the mature leaf powder diets. There were plant-light treatment by larval thermal regime interactions. For example, at 26:15 °C , plant-light treatment had no effect on stadium duration, but at 21:10 °C, stadium duration was prolonged with the full sunlight-new leaf diet compared with the shaded-new leaf diet. In a second diet experiment, we examined the interactive effects of protein and some tomato allelochemicals (rutin, chlorogenic acid and tomatine) on the performance of caterpillars. There were food quality by thermal regime interactions. For instance, at 26:15 °C , neither protein nor allelochemical concentration influenced stadium duration, whereas at 21:10 °C, stadium duration was prolonged with the low protein-high allelochemical diet, which simulated full sunlight leaves. In sum, light availability to plants affected defensive chemistry and protein concentration. The difference in food quality was great enough to influence the growth of a specialist insect herbivore, but the effects were temperature-dependent.     

Response of five insect herbivores to multiple allelochemicals under fluctuating temperatures

Analysis of the combined effects of allelochemicals on insect herbivores is useful because there may be adverse additive or even synergistic effects. Analysis of the simultaneous effects of temperature and alleochemicals is also necessary because these factors may interact. We examined the effects of three allelochemicals found in tomato (chlorogenic acid, rutin and tomatine) and thermal regime (21:10 °C and 26:15 °C, representing spring and summer respectively) on five insect herbivores (a Solanaceae specialist, Manduca sexta, and the polyphagous Heliothis virescens, Pseudoplusia includens, Spodoptera frugiperda and Trichoplusia ni). There were allelochemical interactions and thermal regime-allelochemical interactions for all species, and so the patterns were complex. In some cases, paired allelochemicals or the combination of three allelochemicals showed adverse additive effects on insect performance. But that was not always the case, and there were only a few examples of synergism. Negative effects of the allelochemicals were sometimes, but not always, damped by the cooler thermal regime. Comparing the growth rates of the five species in this study with those of a previous study (a total of seven species) revealed five patterns. For two of three pairs of closely-related species, the paired species had distinctly different patterns. For example, for H. virescens, tomatine prevented development and chlorogenic acid slowed growth, whereas for Helicoverpa zea, tomatine just slowed growth and the phenolics had little effect. The specialist Manduca sexta had a pattern that was midway between patterns of the generalists; it was not the most tolerant of the allelochemicals.     

Isolation of tomatine from cultured excised roots and callus tissues of tomato

Tomatine was present in cultured excised tomato roots but in lower concentrations than in seedling radicles of the same age. The alkaloid was not detected in 'spent' root medium. Newly-initiated callus cultures of hypocotyl, radicle and cotyledon origin produced roots, and tomatine was isolated from both roots and callus. Roots contained more tomatine than callus, but neither contained as much as the organ explants from which the cultures were initiated. The number of roots produced decreased with time, as did also the tomatine content of the callus tissues. After 447 days, when no organized structures were produced by callus cultures, tomatine was not detected. An established hypocotyl callus contained small amounts of tomatine when grown on certain nutrient media, but a chlorophyllous sub-isolate of this callus did not produce detectable quantities of the alkaloid. Tomatine was not detected in an established root callus isolate or in suspension cultures initiated from established, tomatine-containing hypocotyl callus.     

A radioligand assay of tomatine

A novel method for the assay of tomatine in plant extracts is described. It is based on precipitation of tomatine with cholesterols-4-¹⁴C and determination of the radioactivity remaining in solution by scintillation counting. Pigments that cause quenching are removed by chromatography on Sephadex LH20.