The role of primary metabolism in plant resistance against herbivory: a study with the native annual Nicotiana attenuata

Pflanzen haben im Laufe einer langen Koevolution mit Herbivoren verschiedene Mechanismen entwickelt, um das Ausmaß und die Auswirkungen von Biomassenschädigung und -verlust durch Herbivorie zu verringern. Direkte und indirekte Verteidigung reduzieren den Schaden an der Pflanze, indem Herbivoren mit mechanischer Abwehr (z. B. Dornen) abgewehrt, mit chemisch-physiologischer Abwehr (Nervengiften wie Nikotin oder verdauungshemmenden Proteinen, Proteaseinhibitoren) in ihrer Entwicklung gestört werden, oder indem die Pflanze durch Abgabe flüchtiger Verbindungen Fraßfeinde oder Parasitoiden von Herbivoren anlockt. Die Untersuchung dieser Mechanismen erfährt heute ein zunehmend starkes Interesse, weniger gut untersucht ist jedoch, inwieweit es Pflanzen möglich ist, nicht nur die Schädigung an sich, sondern auch die negativen Auswirkungen eines bereits eingetretenen Schadensereignisses abzumildern, oder sogar ein bestimmtes Ausmaß an Schaden ohne (Darwin´sche) Fitnessverluste hinzunehmen. Diese - im Allgemeinen als Toleranz bezeichnete - Pflanzenantwort kann auf unterschiedlichen Mechanismen beruhen, die größtenteils nur theoretisch beschrieben sind

The role of primary metabolism in plant resistance against herbivory: a study with the native annual Nicotiana attenuata

Pflanzen haben im Laufe einer langen Koevolution mit Herbivoren verschiedene Mechanismen entwickelt, um das Ausmaß und die Auswirkungen von Biomassenschädigung und -verlust durch Herbivorie zu verringern. Direkte und indirekte Verteidigung reduzieren den Schaden an der Pflanze, indem Herbivoren mit mechanischer Abwehr (z. B. Dornen) abgewehrt, mit chemisch-physiologischer Abwehr (Nervengiften wie Nikotin oder verdauungshemmenden Proteinen, Proteaseinhibitoren) in ihrer Entwicklung gestört werden, oder indem die Pflanze durch Abgabe flüchtiger Verbindungen Fraßfeinde oder Parasitoiden von Herbivoren anlockt. Die Untersuchung dieser Mechanismen erfährt heute ein zunehmend starkes Interesse, weniger gut untersucht ist jedoch, inwieweit es Pflanzen möglich ist, nicht nur die Schädigung an sich, sondern auch die negativen Auswirkungen eines bereits eingetretenen Schadensereignisses abzumildern, oder sogar ein bestimmtes Ausmaß an Schaden ohne (Darwin´sche) Fitnessverluste hinzunehmen. Diese - im Allgemeinen als Toleranz bezeichnete - Pflanzenantwort kann auf unterschiedlichen Mechanismen beruhen, die größtenteils nur theoretisch beschrieben sind

Oviposition by Spodoptera exigua on Solanum dulcamara Alters the Plant’s Response to Herbivory and Impairs Larval Performance

Plant resistance traits against insect herbivores are extremely plastic. Plants respond not only to the herbivory itself, but also to oviposition by herbivorous insects. How prior oviposition affects plant responses to larval herbivory is largely unknown. Combining bioassays and defense protein activity assays with microarray analyses and metabolite profiling, we investigated the impact of preceding oviposition on the interaction of Solanum dulcamara with the generalist lepidopteran herbivore Spodoptera exigua at the levels of the plant’s resistance, transcriptome and metabolome. We found that oviposition increased plant resistance to the subsequent feeding larvae. While constitutive and feeding-induced levels of defensive protease inhibitor activity remained unaffected, pre-exposure to eggs altered S. dulcamara’s transcriptional and metabolic response to larval feeding in leaves local and systemic to oviposition. In particular, genes involved in phenylpropanoid metabolism were more strongly expressed in previously oviposited plants, which was reflected by reciprocal changes of primary metabolites upstream and within these pathways. Our data highlight that plants integrate signals from non-threatening life stages of their natural enemies to optimize their response when they become actually attacked. The observed transcriptional and metabolic reshaping of S. dulcamara’s response to S. exigua herbivory suggests a role of phenylpropanoids in oviposition-primed plant resistance

Induced, Imprinted, and Primed Responses to Changing Environments: Does Metabolism Store and Process Information?

Metabolism is the system layer that determines growth by the rate of matter uptake and conversion into biomass. The scaffold of enzymatic reaction rates drives the metabolic network in a given physico-chemical environment. In response to the diverse environmental stresses, plants have evolved the capability of integrating macro- and micro-environmental events to be prepared, i.e., to be primed for upcoming environmental challenges. The hierarchical view on stress signaling, where metabolites are seen as final downstream products, has recently been complemented by findings that metabolites themselves function as stress signals. We present a systematic concept of metabolic responses that are induced by environmental stresses and persist in the plant system. Such metabolic imprints may prime metabolic responses of plants for subsequent environmental stresses. We describe response types with examples of biotic and abiotic environmental stresses and suggest that plants use metabolic imprints, the metabolic changes that last beyond recovery from stress events, and priming, the imprints that function to prepare for upcoming stresses, to integrate diverse environmental stress histories. As a consequence, even genetically identical plants should be studied and understood as phenotypically plastic organisms that continuously adjust their metabolic state in response to their individually experienced local environment. To explore the occurrence and to unravel functions of metabolic imprints, we encourage researchers to extend stress studies by including detailed metabolic and stress response monitoring into extended recovery phases

Reverse Genetics in Ecological Research

By precisely manipulating the expression of individual genetic elements thought to be important for ecological performance, reverse genetics has the potential to revolutionize plant ecology. However, untested concerns about possible side-effects of the transformation technique, caused by Agrobacterium infection and tissue culture, on plant performance have stymied research by requiring onerous sample sizes. We compare 5 independently transformed Nicotiana attenuata lines harboring empty vector control (EVC) T-DNA lacking silencing information with isogenic wild types (WT), and measured a battery of ecologically relevant traits, known to be important in plant-herbivore interactions: phytohormones, secondary metabolites, growth and fitness parameters under stringent competitive conditions, and transcriptional regulation with microarrays. As a positive control, we included a line silenced in trypsin proteinase inhibitor gene (TPI) expression, a potent anti-herbivore defense known to exact fitness costs in its expression, in the analysis. The experiment was conducted twice, with 10 and 20 biological replicates per genotype. For all parameters, we detected no difference between any EVC and WT lines, but could readily detect a fitness benefit of silencing TPI production. A statistical power analyses revealed that the minimum sample sizes required for detecting significant fitness differences between EVC and WT was 2–3 orders of magnitude larger than the 10 replicates required to detect a fitness effect of TPI silencing. We conclude that possible side-effects of transformation are far too low to obfuscate the study of ecologically relevant phenotypes

A Signaling Protease Required for Melanization in Drosophila Affects Resistance and Tolerance of Infections

Organisms evolve two routes to surviving infections—they can resist pathogen growth (resistance) and they can endure the pathogenesis of infection (tolerance). The sum of these two properties together defines the defensive capabilities of the host. Typically, studies of animal defenses focus on either understanding resistance or, to a lesser extent, tolerance mechanisms, thus providing little understanding of the relationship between these two mechanisms. We suggest there are nine possible pairwise permutations of these traits, assuming they can increase, decrease, or remain unchanged in an independent manner. Here we show that by making a single mutation in the gene encoding a protease, CG3066, active in the melanization cascade in Drosophila melanogaster, we observe the full spectrum of changes; these mutant flies show increases and decreases in their resistance and tolerance properties when challenged with a variety of pathogens. This result implicates melanization in fighting microbial infections and shows that an immune response can affect both resistance and tolerance to infections in microbe-dependent ways. The fly is often described as having an unsophisticated and stereotypical immune response where single mutations cause simple binary changes in immunity. We report a level of complexity in the fly's immune response that has strong ecological implications. We suggest that immune responses are highly tuned by evolution, since selection for defenses that alter resistance against one pathogen may change both resistance and tolerance to other pathogens

Tracing Personalized Health Curves during Infections

By concentrating on the relationship between health and microbe number over the course of infections, most pathogenic and mutualistic infections can be summarized by a small alphabet of curves, which has implications not only for basic research but for how we might treat patients

Tobacco Rattle Virus Vector: A Rapid and Transient Means of Silencing Manduca sexta Genes by Plant Mediated RNA Interference

Background: RNAi can be achieved in insect herbivores by feeding them host plants stably transformed to express double stranded RNA (dsRNA) of selected midgut-expressed genes. However, the development of stably transformed plants is a slow and laborious process and here we developed a rapid, reliable and transient method. We used viral vectors to produce dsRNA in the host plant Nicotiana attenuata to transiently silence midgut genes of the plant’s lepidopteran specialist herbivore, Manduca sexta. To compare the efficacy of longer, undiced dsRNA for insect gene silencing, we silenced N. attenuata’s dicer genes (NaDCL1- 4) in all combinations in a plant stably transformed to express dsRNA targeting an insect gene. Methodology/Principal Findings: Stable transgenic N. attenuata plants harboring a 312 bp fragment of MsCYP6B46 in an inverted repeat orientation (ir-CYP6B46) were generated to produce CYP6B46 dsRNA. After consuming these plants, transcripts of CYP6B46 were significantly reduced in M. sexta larval midguts. The same 312 bp cDNA was cloned in an antisense orientation into a TRV vector and Agro-infiltrated into N. attenuata plants. When larvae ingested these plants, similar reductions in CYP6B46 transcripts were observed without reducing transcripts of the most closely related MsCYP6B45. We used this transient method to rapidly silence the expression of two additional midgut-expressed MsCYPs. CYP6B46 transcripts were further reduced in midguts, when the larvae fed on ir-CYP6B46 plants transiently silenced for tw

Metabolomic Plasticity in GM and Non-GM Potato Leaves in Response to Aphid Herbivory and Virus Infection

An important aspect of ecological safety of genetically modified (GM) plants is the evaluation of unintended effects on plant–insect interactions. These interactions are to a large extent influenced by the chemical composition of plants. This study uses NMR-based metabolomics to establish a baseline of chemical variation to which differences between a GM potato line and its parent cultivar are compared. The effects of leaf age, virus infection, and aphid herbivory on plant metabolomes were studied. The metabolome of the GM line differed from its parent only in young leaves of noninfected plants. This effect was small when compared to the baseline. Consistently, aphid performance on excised leaves was influenced by leaf age, while no difference in performance was found between GM and non-GM plants. The metabolomic baseline approach is concluded to be a useful tool in ecological safety assessment