Meredith C. Schuman

Jasmonate and ppHsystemin regulate key malonylation steps in the biosynthesis of 17-hydroxygeranyllinalool diterpene glycosides, an abundant and effective direct defense against herbivores in Nicotiana attenuata

This study describes the biosynthetic dynamics of diterpene glycosides abundant in a wild tobacco and highlights glycosylation and malonylation as key biosynthetic steps producing the diversity of compounds observed. It shows that plants silenced in diterpene glycoside production suffered more damage from herbivores in their natural habitat and were higher-quality food for a specialist herbivore. We identified 11 17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) that occur in concentrations equivalent to starch (mg/g fresh mass) in aboveground tissues of coyote tobacco (Nicotiana attenuata) and differ in their sugar moieties and malonyl sugar esters (0-2). Concentrations of HGL-DTGs, particularly malonylated compounds, are highest in young and reproductive tissues. Within a tissue, herbivore elicitation changes concentrations and biosynthetic kinetics of individual compounds. Using stably transformed N. attenuata plants silenced in jasmonate production and perception, or production of N. attenuata Hyp-rich glycopeptide systemin precursor by RNA interference, we identified malonylation as the key biosynthetic step regulated by herbivory and jasmonate signaling. We stably silenced N. attenuata geranylgeranyl diphosphate synthase (ggpps) to reduce precursors for the HGL-DTG skeleton, resulting in reduced total HGL-DTGs and greater vulnerability to native herbivores in the field. Larvae of the specialist tobacco hornworm (Manduca sexta) grew up to 10 times as large on ggpps silenced plants, and silenced plants suffered significantly more damage from herbivores in N. attenuatas native habitat than did wild-type plants. We propose that high concentrations of HGL-DTGs effectively defend valuable tissues against herbivores and that malonylation may play an important role in regulating the distribution and storage of HGL-DTGs in plants.

Herbivory-induced volatiles function as defenses increasing fitness of the native plant Nicotiana attenuata in nature

From an herbivores first bite, plants release herbivory-induced plant volatiles (HIPVs) which can attract enemies of herbivores. However, other animals and competing plants can intercept HIPVs for their own use, and it remains unclear whether HIPVs serve as an indirect defense by increasing fitness for the emitting plant. In a 2-year field study, HIPV-emitting N. attenuata plants produced twice as many buds and flowers as HIPV-silenced plants, but only when native Geocoris spp. predators reduced herbivore loads (by 50%) on HIPV-emitters. In concert with HIPVs, plants also employ antidigestive trypsin protease inhibitors (TPIs), but TPI-producing plants were not fitter than TPI-silenced plants. TPIs weakened a specialist herbivores behavioral evasive responses to simulated Geocoris spp. attack, indicating that TPIs function against specialists by enhancing indirect defense. DOI: http://dx.doi.org/10.7554/eLife.00007.001

Polymorphism in jasmonate signaling partially accounts for the variety of volatiles produced by Nicotiana attenuata plants in a native population

Herbivore- and jasmonate-induced volatile organic compounds (VOCs), which mediate indirect defense, must provide reliable information for predators that frequently learn to associate their release with feeding herbivores. Yet little is known about variation of these cues within populations of native plants, on a scale encountered by predators. We examined variation in herbivore-elicited VOC emissions and patterns of herbivore-induced jasmonate signaling from accessions of Nicotiana attenuata co-occurring in a native population. VOC emissions elicited by herbivore oral secretions (OS) and by methyl jasmonate (MJ) were characterized using gas chromatography-mass spectrometry (GC-MS), high-resolution two-dimensional gas chromatography-time-of-flight mass spectrometry (GCxGC-ToF-MS) and micro-hydrolysis and micro-hydrogenation reactions. Accessions varied in emissions of abundant (trans-alpha-bergamotene, alpha-duprezianene, trans-beta-ocimene, and cis-3-hexenol) and total detectable VOCs, as well as the accumulation of jasmonates, the jasmonate antagonist salicylic acid (SA), abscisic acid (ABA) and jasmonate signaling-related transcripts after OS elicitation. Yet MJ treatment exacerbated differences in VOC emission, suggesting that much variation in VOC emission is caused by processes downstream of jasmonate signaling. Co-occurring N. attenuata plants emit different VOCs following simulated herbivore elicitation as a result in part of differences in jasmonate production and responsiveness, which could reduce the effectiveness of induced indirect defense.

The layers of plant responses to insect herbivores

Plants collectively produce hundreds of thousands of specialized metabolites that are not required for growth or development. Each species has a qualitatively unique profile, with variation among individuals, growth stages, and tissues. By the 1950s, entomologists began to recognize the supreme importance of these metabolites in shaping insect herbivore communities. Plant defense theories arose to address observed patterns of variation, but provided few testable hypotheses because they did not distinguish clearly among proximate and ultimate causes. Molecular plant-insect interaction research has since revealed the sophistication of plant metabolic, developmental, and signaling networks. This understanding at the molecular level, rather than theoretical predictions, has driven the development of new hypotheses and tools and pushed the field forward. We reflect on the utility of the functional perspective provided by the optimal defense theory, and propose a conceptual model of plant defense as a series of layers each at a different level of analysis, illustrated by advances in the molecular ecology of plant-insect interactions.

A comparison of two Nicotiana attenuata accessions reveals large differences in signaling induced by oral secretions of the specialist herbivore Manduca sexta

Genetic variation within and among populations provides the raw material for evolution. Although many studies describe inter- and intraspecific variation of defensive metabolites, little is known about variation among plant populations within early signaling responses elicited by herbivory or by herbivore oral secretions (OS) introduced into wounds during feeding. In this study, we compare the OS-elicited early responses as well as the antiherbivore defensive metabolites in two accessions of the wild tobacco Nicotiana attenuata and show that, compared with an accession collected from Utah, an Arizona accession has lower herbivore-elicited activity of the salicylic acid-induced protein kinase, an important mitogen-activated protein kinase involved in herbivore resistance. These differences in salicylic acid-induced protein kinase activity were associated with substantially different levels of OS-elicited jasmonic acid, jasmonic acid-isoleucine conjugate, and ethylene bursts. Gene expression level polymorphism (ELP) determines phenotypic variation among populations, and we found the two accessions to have significantly different ELPs in the genes involved in early signaling responses to herbivory. In addition, we found differences between the Utah and the Arizona accessions in the concentrations of several secondary metabolites that contribute to N. attenuatas direct and indirect defenses. This study demonstrates significant natural variation in regulatory elements that mediate plant responses to herbivore attack, highlighting the role of ELP in producing a diversity of plant defense phenotypes.

Silencing jasmonate signalling and jasmonate-mediated defenses reveals different survival strategies between two Nicotiana attenuata accessions

To determine the impact of genotypic variation in secondary metabolite production on antiherbivore resistance and plant fitness, we genetically silenced biosynthetic genes for nicotine, trypsin proteinase inhibitors (TPI), and jasmonate (JA) production in two accessions of Nicotiana attenuata: one from Utah (UT) which responds to herbivory with JA‐induced nicotine and TPI production, and one from Arizona (AZ) which is TPI‐deficient but also produces JA‐induced nicotine. Transient silencing of JA biosynthesis increased Manduca sexta larval growth on wild type (WT) plants of both accessions, but not on TPI‐deficient UT or nicotine‐deficient AZ lines, demonstrating that JA‐mediated resistance to M. sexta requires TPIs in the UT and nicotine in the naturally TPI‐deficient AZ accession. When transplanted into a native UT population, AZ and UT plants, rendered equally able or unable to produce nicotine and TPIs by stable transformation, received significantly different levels of herbivory. Both accessions differed in their resistance depending on the type of herbivores: resistance to rare, voracious herbivores (Saltatoria and Mammalia) was greater in AZ than UT lines, and dependent on nicotine production, while resistance to small, abundant herbivores (Coleoptera and Thysanoptera) was greater in UT lines, and dependent on TPI production. AZ lines produced more flowers and seed capsules than UT lines independently of TPI production costs. This fitness advantage was lost when accessions did not produce nicotine. We conclude that these two accessions have developed different survival strategies and thus differ in the cost‐benefit functions of their JA‐mediated defences.

MAPK signaling - a key element in plant defense response to insects

Insects have long been the most abundant herbivores, and plants have evolved sophisticated mechanisms to defend against their attack. In particular, plants can perceive specific patterns of tissue damage associated with insect herbivory. Some plant species can perceive certain elicitors in insect oral secretions (OS) that enter wounds during feeding, and rapidly activate a series of intertwined signaling pathways to orchestrate the biosynthesis of various defensive metabolites. Mitogen‐activated protein kinases (MAPKs), common to all eukaryotes, are involved in the orchestration of many cellular processes, including development and stress responses. In plants, at least two MAPKs, salicylic acid‐induced protein kinase (SIPK) and wound‐induced protein kinase (WIPK), are rapidly activated by wounding or insect OS; importantly, genetic studies using transgenic or mutant plants impaired in MAPK signaling indicated that MAPKs play critical roles in regulating the herbivory‐induced dynamics of phytohormones, such as jasmonic acid, ethylene and salicylic acid, and MAPKs are also required for transcriptional activation of herbivore defense‐related genes and accumulation of defensive metabolites. In this review, we summarize recent developments in understanding the functions of MAPKs in plant resistance to insect herbivores.

Plant defense phenotypes determine the consequences of volatile emission for individuals and neighbors

Plants are at the trophic base of terrestrial ecosystems, and the diversity of plant species in an ecosystem is a principle determinant of community structure. This may arise from diverse functional traits among species. In fact, genetic diversity within species can have similarly large effects. However, studies of intraspecific genetic diversity have used genotypes varying in several complex traits, obscuring the specific phenotypic variation responsible for community-level effects. Using lines of the wild tobacco Nicotiana attenuata genetically altered in specific well-characterized defense traits and planted into experimental populations in their native habitat, we investigated community-level effects of trait diversity in populations of otherwise isogenic plants. We conclude that the frequency of defense traits in a population can determine the outcomes of these traits for individuals. Furthermore, our results suggest that some ecosystem-level services afforded by genetically diverse plant populations could be recaptured in intensive monocultures engineered to be functionally diverse. DOI: http://dx.doi.org/10.7554/eLife.04490.001

The Sesquiterpenes(E)-ß-Farnesene and (E)-α-Bergamotene Quench Ozone but Fail to Protect the Wild Tobacco Nicotiana attenuata from Ozone, UVB, and Drought Stresses

Among the terpenes, isoprene (C5) and monoterpene hydrocarbons (C10) have been shown to ameliorate abiotic stress in a number of plant species via two proposed mechanisms: membrane stabilization and direct antioxidant effects. Sesquiterpene hydrocarbons (C15) not only share the structural properties thought to lend protective qualities to isoprene and monoterpene hydrocarbons, but also react rapidly with ozone, suggesting that sesquiterpenes may similarly enhance tolerance of abiotic stresses. To test whether sesquiterpenes protect plants against ozone, UVB light, or drought, we used transgenic lines of the wild tobacco Nicotiana attenuata. The transgenic plants expressed a maize terpene synthase gene (ZmTPS10) which produced a blend of (E)-ß-farnesene and (E)-α-bergamotene, or a point mutant of the same gene (ZmTPS10M) which produced (E)-ß-farnesene alone,. (E)-ß-farnesene exerted a local, external, and transient ozone-quenching effect in ozone-fumigated chambers, but we found no evidence that enhanced sesquiterpene production by the plant inhibited oxidative damage, or maintained photosynthetic function or plant fitness under acute or chronic stress. Although the sesquiterpenes (E)-ß-farnesene and (E)-α-bergamotene might confer benefits under intermittent heat stress, which was not tested, any roles in relieving abiotic stress may be secondary to their previously demonstrated functions in biotic interactions.

Chemical Classification of the Essential Oils of the Iranian Salvia Species in Comparison with Their Botanical Taxonomy

The essential oils of eight Salvia species collected from different localities in Iran were analyzed by gas chromatography/mass spectrometry (GC/MS). The analytical results were compared with those previously published for related Iranian sage species in order to identify chemical markers for these species. Salvia eremophila, S. hypoleuca, and S. reuteriana are endemic, while S. atropatana, S. chloroleuca, S. santolinifolia, S. aegyptiaca, and S. macrosiphon also grow wild in neighboring countries. We categorized the Iranian Salvia species into four main chemotypes according to their essential‐oil constituents: those which are dominated by 1) monoterpenes, 2) mono‐ and sesquiterpenes, or 3) sesquiterpenes as the major constituents, and 4) those containing low‐molecular‐weight acids, aldehydes, and esters, and green‐leaf volatiles (GLVs). Likely due to the chemical diversity of different Salvia chemotypes, this categorization was supported by principal component analysis (PCA) for the group sampled here, but not for the values reported in the literature. We identified the following chemical markers: α‐pinene, β‐pinene, 1,8‐cineol, linalool, and borneol in monoterpene‐rich species, or β‐caryophyllene, germacrene D, bicyclogermacrene, spathulenol, and caryophyllene oxide in sesquiterpene‐rich species. Among these, α‐pinene, β‐caryophyllene, and germacrene D are the most common and abundant in the Salvia species investigated. In accordance with their close biological taxonomy, the chemical similarity of the essential oils of S. santolinifolia and S. eremophila is so high that we may consider them chemically identical.