Insect herbivory on plants is a complex incident consisting of at least two different aspects, mechanical damage and chemical factors. Only the combination of both is able to induce the respective plant defenses. Thus, diverse plant species emit volatile organic compounds (VOCs) in response to herbivory (HW), whereas mechanical damage inflicted as single wounding event (MD) does not induce increased VOC emissions. In contrast, a robotic worm (MecWorm, MW) allowed demonstrating that continuous mechanical damage is sufficient to induce volatile emission in Lima bean. However, the induced VOC blends remain characteristic for the respective stimulus. In order to identify putative differences in plant signaling leading to defenses, we compared time courses of early signals induced by wounding in Lima bean. Neither MD nor MW alone was able to induce plasma membrane (V (m)) depolarization, as observed after Spodoptera littoralis HW, but V (m) depolarization occurred in both treatments when used in combination with herbivore-derived oral secretions. A significant increase in cytosolic Ca(2+) concentrations was observed only after HW, whereas MD and MW did not affect this second messenger. H(2)O(2) was generated within 2-3 h after leaf damage by HW and MW, whereas MD induced only half of the H(2)O(2) levels compared to the other treatments. Both HW and MW induced a marked accumulation of NO, but with distinct temporal patterns. NO production after MD followed the same trend but reached significantly lower values. The results indicate that chemical signals from the herbivores are responsible for the induction of the earliest signaling events. These changes appear to be characteristic for the reaction to herbivory.

Background Ginkgo biloba (Ginkgoaceae) is one of the most ancient living seed plants and is regarded as a living fossil. G. biloba has a broad spectrum of resistance or tolerance to many pathogens and herbivores because of the presence of toxic leaf compounds. Little is known about early and late events occurring in G. biloba upon herbivory. The aim of this study was to assess whether herbivory by the generalist Spodoptera littoralis was able to induce early signaling and direct defense in G. biloba by evaluating early and late responses.Methodology/Principal FindingsEarly and late responses in mechanically wounded leaves and in leaves damaged by S. littoralis included plasma transmembrane potential (Vm) variations, time-course changes in both cytosolic calcium concentration ([Ca2+]cyt) and H2O2 production, the regulation of genes correlated to terpenoid and flavonoid biosynthesis, the induction of direct defense compounds, and the release of volatile organic compounds (VOCs). The results show that G. biloba responded to hebivory with a significant Vm depolarization which was associated to significant increases in both [Ca2+]cyt and H2O2. Several defense genes were regulated by herbivory, including those coding for ROS scavenging enzymes and the synthesis of terpenoids and flavonoids. Metabolomic analyses revealed the herbivore-induced production of several flavonoids and VOCs. Surprisingly, no significant induction by herbivory was found for two of the most characteristic G. biloba classes of bioactive compounds; ginkgolides and bilobalides.Conclusions/SignificanceBy studying early and late responses of G. biloba to herbivory, we provided the first evidence that this “living fossil” plant responds to herbivory with the same defense mechanisms adopted by the most recent angiosperms.

Conservation biological control aims to enhance natural enemy populations in crop habitats, e.g. by providing flowering plants as food resources. Suitable flower species must enhance the survival and fecundity of natural enemies but in addition they also need to be highly attractive and thus frequently visited. To date, few examples exist that have considered both criteria. In this study, we tested the effects of the flowering plants alyssum (Lobularia maritima), buckwheat (Fagopyrum esculentum), French marigold (Tagetes patula) and sweet basil (Ocimum basilicum) on the fecundity and olfactory attractiveness of the egg parasitoid Trissolcus basalis, an important biological control agent of the stink bug Nezara viridula. Our results showed that access to buckwheat and basil flowers increased the parasitoid offspring. However, in olfactometer experiments where T. basalis was allowed to choose between flowering and non-flowering plants, only buckwheat floral scent was attractive. Headspace analyses of the odour emitted by the four plant species revealed very distinct profiles with little overlap in compounds. Buckwheat floral scent was characterized by an unpleasant smell for the human nose due to the presence of short-chain carboxylic acids. Headspace extracts of buckwheat flowers and a blend of six buckwheat plant volatiles consisting of butanoic, 2-methylbutanoic, 3-methylbutanoic and pentanoic acids, (Z)-3-hexenyl acetate and α-farnesene were significantly attractive in olfactometer bioassays. Furthermore, electrophysiological experiments showed most of these compounds elicited significant responses in T. basalis antennae. Integrating chemo-ecological methods into conservation biological control allowed us to identify a potential resource plant and attractive compounds for field studies

We investigated the role of polyamines (PAs) in lima bean (Phaseolus lunatus) leaves on the production of herbivorous mite (Tetranychus urticae)-induced plant volatiles that attract carnivorous natural enemies of the herbivores. To do this, we focused on the effects of the exogenous PAs [cadaverine, putrescine, spermidine and spermine (Spm)] on the production of volatiles, H(2)O(2) and jasmonic acid (JA) and the levels of defensive genes, cytosolic calcium and reactive oxygen species (ROS). Among the tested PAs, Spm was the most active in inducing the production of volatile terpenoids known to be induced by T. urticae. An increase in JA levels was also found after Spm treatment, indicating that Spm induces the biosynthesis of JA, which has been shown elsewhere to regulate the production of some volatile terpenoids. Further, treatment with JA and Spm together resulted in greater volatile emission than that with JA alone. In a Y-tube olfactometer, leaves treated with Spm + JA attracted more predatory mites (Phytoseiulus persimilis) than those treated with JA alone. After treatment with Spm + JA, no effects were found on the enzyme activity of polyamine oxidase and copper amine oxidase. However, induction of calcium influx and ROS production, and increased enzyme activities and gene expression for NADPH oxidase complex, superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and glutathione peroxidase were found after treatment with Spm + JA. These results indicate that Spm plays an important role in the production of T. urticae-induced lima bean leaf volatiles.