Chemicals on plant surfaces as a heretofore unrecognized,but ecologically informative,class for investigations into plant defence

Plants produce and utilize a great diversity of chemicals for a variety of physiological and ecological purposes. Many of these chemicals defend plants against herbivores, pathogens and competitors. The location of these chemicals varies within the plant, some are located entirely within plant tissues, others exist in the air‐ (or water‐) space around plants, and still others are secreted onto plant surfaces as exudates. I argue herein that the location of a given defensive chemical has profound implications for its ecological function; specifically, I focus on the characteristics of chemical defences secreted onto plant surfaces. Drawing from a broad literature encompassing ecology, evolution, taxonomy and physiology, I found that these external chemical defences (ECDs) are common and widespread in plants and algae; hundreds of examples have been detailed, yet they are not delineated as a separate class from internal chemical defences (ICDs). I propose a novel typology for ECDs and, using existing literature, explore the ecological consequences of the hypothesized unique characteristics of ECDs. The axis of total or proportional investment in ECDs versus ICDs should be considered as one axis of investment by a plant, in the same way as quantitative versus qualitative chemical defences or induced versus constitutive defences is considered. The ease of manipulating ECDs in many plant systems presents a powerful tool to help test plant defence theory (e.g. optimal defence). The framework outlined here integrates various disciplines of botany and ecology and suggests a need for further examinations of exudates in a variety of contexts, as well as recognition of the effects of within‐plant localization of defences.     

Synergies and trade-offs between insect and pathogen resistance in maize leaves and roots

Determining links between plant defence strategies is important to understand plant evolution and to optimize crop breeding strategies. Although several examples of synergies and trade-offs between defence traits are known for plants that are under attack by multiple organisms, few studies have attempted to measure correlations of defensive strategies using specific single attackers. Such links are hard to detect in natural populations because they are inherently confounded by the evolutionary history of different ecotypes. We therefore used a range of 20 maize inbred lines with considerable differences in resistance traits to determine if correlations exist between leaf and root resistance against pathogens and insects. Aboveground resistance against insects was positively correlated with the plant's capacity to produce volatiles in response to insect attack. Resistance to herbivores and resistance to a pathogen, on the other hand, were negatively correlated. Our results also give first insights into the intraspecific variability of root volatiles release in maize and its positive correlation with leaf volatile production. We show that the breeding history of the different genotypes (dent versus flint) has influenced several defensive parameters. Taken together, our study demonstrates the importance of genetically determined synergies and trade-offs for plant resistance against insects and pathogens.     

Root exudates and plant secondary metabolites of different plants enhance polychlorinated biphenyl degradation by rhizobacteria

Polychlorinated biphenyls (PCBs) are toxic and persistent compounds that are difficult to break down and biodegrade. Plant secondary metabolites (PSMs) on root exudates can act as inducers of the biphenyl catabolic pathway, enhancing PCB biodegradation. In this study, the authors evaluated the effect of root exudates and PSMs obtained from Avena sativa, Brachiaria decumbens, Medicago sativa, and Brassica juncea on the biodegradation of PCB 44, PCB 66, PCB 118, PCB 138, PCB 153, PCB 170, and PCB 180 by a microbial consortium isolated from the rhizosphere of plants grown on soil contaminated with Aroclor 1260. Microorganisms were identified as Pseudomonas sp. and Stenotrophomonas sp. based on their 16S rRNA sequence. The plant root exudates increased the degradation percentage of PCB 44, PCB 66, and PCB 118, which were used as carbon source by the microorganisms. Flavanone, flavone, isoflavone, 7-hydroxyflavanone, 7-hydroxyflavone, and 6-hydroxyflavone were the PSMs identified in the root exudates, which increased the degradation percentage of all seven PCB congeners; they were also used as growth substrates by microbial consortium. These results showed the importance of the interaction between plants and microorganisms for achieving the removal of persistent pollutants such as PCBs from soil.     

Herbivore‐induced plant responses in Brassica oleracea prevail over effects of constitutive resistance and result in enhanced herbivore attack

1. Plant responses to herbivore attack may have community‐wide effects on the composition of the plant‐associated insect community. Thereby, plant responses to an early‐season herbivore may have profound consequences for the amount and type of future attack. 2. Here we studied the effect of early‐season herbivory by caterpillars of Pieris rapae on the composition of the insect herbivore community on domesticated Brassica oleracea plants. We compared the effect of herbivory on two cultivars that differ in the degree of susceptibility to herbivores to analyse whether induced plant responses supersede differences caused by constitutive resistance. 3. Early‐season herbivory affected the herbivore community, having contrasting effects on different herbivore species, while these effects were similar on the two cultivars. Generalist insect herbivores avoided plants that had been induced, whereas these plants were colonised preferentially by specialist herbivores belonging to both leaf‐chewing and sap‐sucking guilds. 4. Our results show that community‐wide effects of early‐season herbivory may prevail over effects of constitutive plant resistance. Induced responses triggered by prior herbivory may lead to an increase in susceptibility to the dominant specialists in the herbivorous insect community. The outcome of the balance between contrasting responses of herbivorous community members to induced plants therefore determines whether induced plant responses result in enhanced plant resistance.     

Leaf trichome responses to herbivory in willows: induction, relaxation and costs

Direct measurement of the carbon (C) 'cost' of mycorrhizas is problematic. Although estimates have been made for arbuscular and ectomycorrhizal symbioses, these are based on incomplete budgets or indirect measurements. Furthermore, the conventional model of unidirectional plant-to-fungus C flux is too simplistic. Net fungus-to-plant C transfer supports seedling establishment in c. 10% of plant species, including most orchids, and bidirectional C flows occur in ectomycorrhiza utilizing soil amino acids. Here, the C cost of mycorrhizas to the green orchid Goodyera repens was determined by measurement of simultaneous bidirectional fluxes of 14C labelled sources using a monoxenic system with the fungus Ceratobasidium cornigerum. Transfer of C from fungus to plant ('up-flow') occurs in the photosynthesizing orchid G. repens (max. 0.06 microg) whereas over five times more current assimilate (min. 0.355 microg) is simultaneously allocated in the reverse direction to the mycorrhizal fungus ('down-flow') after 8 d. Carbon is transferred rapidly, being detected in plant-fungal respiration within 31 h of labelling. This study provides the most complete C budget for an orchid-mycorrhizal symbiosis, and clearly shows net plant-to-fungus C flux. The rapidity of bidirectional C flux is indicative of dynamic transfer at an interfacial apoplast as opposed to reliance on digestion of fungal pelotons.     

Elephant herbivory of knob-thorn (Senegalia nigrescens) and ivory palm (Hyphaene petersiana) in Bwabwata National Park,Caprivi, Namibia: The role of ivory palm as a biotic refuge

African elephants have major impacts on vegetation, particularly at high densities. Knob-thorns (Senegalia nigrescens) are typically ring-barked by elephant, and high levels of mortality are common at high elephant densities. Our study aimed to test whether ivory palm clusters (Hyphaene petersiana) form a biotic refuge for knob-thorn against elephant herbivory. We measured the density, damage and mortality of knob-thorns in sites differing according to ivory palm presence and elephant density, and thus, the probability of knob-thorn encounter by elephants. The site with palms and low elephant density, had a high density of knob-thorns, but lower proportions of damaged and dead trees, than sites without palms but with similar or higher elephant density. In the former, knob-thorns were associated with palm clusters, particularly saplings and young adults. In this site, low proportions of damaged and dead knob-thorns were recorded in palm clusters, compared with outside clusters, and to those in the other sites. Our study also showed that juvenile palms which protected knob-thorns, suffered low mortality in contrast to subadult palms. We have no evidence but implicate elephants and suggest that in palm clusters, subadult palms are more accessible to elephants than knob-thorns because of the different methods of utilisation.     

A specialist root herbivore exploits defensive metabolites to locate nutritious tissues

The most valuable organs of plants are often particularly rich in essential elements, but also very well defended. This creates a dilemma for herbivores that need to maximise energy intake while minimising intoxication. We investigated how the specialist root herbivore Diabrotica virgifera solves this conundrum when feeding on wild and cultivated maize plants. We found that crown roots of maize seedlings were vital for plant development and, in accordance, were rich in nutritious primary metabolites and contained higher amounts of the insecticidal 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and the phenolic compound chlorogenic acid. The generalist herbivores Diabrotica balteata and Spodoptera littoralis were deterred from feeding on crown roots, whereas the specialist D. virgifera preferred and grew best on these tissues. Using a 1,4-benzoxazin-3-one-deficient maize mutant, we found that D. virgifera is resistant to DIMBOA and other 1,4-benzoxazin-3-ones and that it even hijacks these compounds to optimally forage for nutritious roots.     

Metabolomics reveals herbivore‐induced metabolites of resistance and susceptibility in maize leaves and roots

Plants respond to herbivory by reprogramming their metabolism. Most research in this context has focused on locally induced compounds that function as toxins or feeding deterrents. We developed an ultra‐high‐pressure liquid chromatography time‐of‐flight mass spectrometry (UHPLC‐TOF‐MS)‐based metabolomics approach to evaluate local and systemic herbivore‐induced changes in maize leaves, sap, roots and root exudates without any prior assumptions about their function. Thirty‐two differentially regulated compounds were identified from Spodoptera littoralis‐infested maize seedlings and isolated for structure assignment by microflow nuclear magnetic resonance (CapNMR). Nine compounds were quantified by a high throughput direct nano‐infusion tandem mass spectrometry/mass spectrometry (MS/MS) method. Leaf infestation led to a marked local increase of 1,3‐benzoxazin‐4‐ones, phospholipids, N‐hydroxycinnamoyltyramines, azealic acid and tryptophan. Only few changes were found in the root metabolome, but 1,3‐benzoxazin‐4‐ones increased in the vascular sap and root exudates. The role of N‐hydroxycinnamoyltyramines in plant–herbivore interactions is unknown, and we therefore tested the effect of the dominating p‐coumaroyltyramine on S. littoralis. Unexpectedly, p‐coumaroyltyramine was metabolized by the larvae and increased larval growth, possibly by providing additional nitrogen to the insect. Taken together, this study illustrates that herbivore attack leads to the induction of metabolites that can have contrasting effects on herbivore resistance in the leaves and roots.     

Intraspecific variation in heritable secondary metabolites and defensive strategies in a relict tree

Aims Key herbivory interaction traits such as plant defensive compounds may differ among populations of a single species due to the spatial variation in herbivore feeding guilds and the strength of the interaction. Moreover, the genealogy of population lineages could represent an additional source of variation interacting with the predominant eco-clinal trends. We tested for the existence of genetically based intraspecific variation in chemical defence profiles across the range of the relict tree Prunus lusitanica L. Additionally, we investigated geographical variation in defence inducibility and tested for the existence of a trade-off between qualitative and quantitative defences.Methods We conducted a greenhouse experiment where 210 plants were grown under a common environment, comprising 10 different populations throughout the distribution range of the species and spanning three separate regions: Iberia, Morocco and Macaronesia. To test for the inducibility of defences, we artificially defoliated plants. Three treatments were established within each population: undamaged, defoliated and sampled after 2 h, and defoliated and sampled after 72 h. The concentration of cyanogenic glycosides (prunasin) and phenolics was determined in leaf samples for all treatments.Important findings Basal levels of cyanogenics and phenolics significantly differed among populations and regions across the range of P. lusitanica, with this variation having a heritable basis. Cyanogenics (prunasin) were significantly higher in ancient Macaronesian populations, while phenolic concentrations were larger in Iberia. The higher cyanogenic levels found in Macaronesia could be a consequence of the known stronger herbivory pressure in the islands than in Iberia or the likely longer coevolutionary history with herbivores in this region. These findings indicate that the geographical variation of key ecological traits such plant chemical defences can be imprinted by phylogeographical signals, particularly in relict species. Regarding defence inducibility, prunasin increased after simulated herbivory whereas phenolics mostly decreased after defoliation. Variation in defence inducibility across populations and regions was evident, although no consistent patterns related to the variation in herbivore feeding guilds were observed, particularly among regions with and without ungulate browsing pressure. Finally, a trade-off among induced levels of qualitative (prunasin) and quantitative (phenolics) defences was detected in one of the defoliated treatments, likely as a result of a stronger resource limitation in damaged plants.     

Glucosinolates in oilseed rape: secondary metabolites that influence interactions with herbivores and their natural enemies

Glucosinolates are sulphur‐containing secondary metabolites characteristic of Brassicaceous plants. Glucosinolate breakdown products, which include isothiocyanates, are released following tissue damage when hydrolytic enzymes act on them. The isothiocyanates have toxic effects on generalist herbivores when they attempt to feed on oilseed rape, Brassica napus, and also function as repellents. However, specialist herbivores such as Brevicoryne brassicae aphids, flea beetles, Psylliodes chrysocephala and the Lepidopteran pest, Pieris rapae, are adapted to the presence of glucosinolates and thrive on plants containing them. They may do this by avoiding tissue damage to prevent the formation of isothiocyanates or by metabolising or tolerating glucosinolates. For many specialist herbivores, the isothiocyanates function as attractants and glucosinolates can even be sequestered for defence against predatory insects. Thus, these herbivores have evolved resistance to host‐plant secondary metabolites and this type of evolutionary history may have given some insects an enhanced ability to adapt to xenobiotics. In an agricultural context, this may make pests better able to evolve resistance to artificially applied pesticides. The effect of increased glucosinolate content in making oilseed rape cultivars more susceptible to specialist pests was highlighted in a seminal article in the Annals of Applied Biology in 1995. This review of the literature considers developments in this area since then.     

Evolution of phenotypic plasticity: Genetic differentiation and additive genetic variation for induced plant defence in wild arugula Eruca sativa

Phenotypic plasticity is the primary mechanism of organismal resilience to abiotic and biotic stress, and genetic differentiation in plasticity can evolve if stresses differ among populations. Inducible defence is a common form of adaptive phenotypic plasticity, and long‐standing theory predicts that its evolution is shaped by costs of the defensive traits, costs of plasticity and a trade‐off in allocation to constitutive versus induced traits. We used a common garden to study the evolution of defence in two native populations of wild arugula Eruca sativa (Brassicaceae) from contrasting desert and Mediterranean habitats that differ in attack by caterpillars and aphids. We report genetic differentiation and additive genetic variance for phenology, growth and three defensive traits (toxic glucosinolates, anti‐nutritive protease inhibitors and physical trichome barriers) as well their inducibility in response to the plant hormone jasmonic acid. The two populations were strongly differentiated for plasticity in nearly all traits. There was little evidence for costs of defence or plasticity, but constitutive and induced traits showed a consistent additive genetic trade‐off within each population for the three defensive traits. We conclude that these populations have evolutionarily diverged in inducible defence and retain ample potential for the future evolution of phenotypic plasticity in defence.     

Florivory by a floriphilic katydid,Phaneroptera brevis,induces changes in a leaf trait in Lantana camara

1. Plants can induce a response when they are attacked by herbivores. Although the induction of responses by herbivory in both flowers and leaves is relatively well studied, whether florivory (feeding of flowers) can also induce responses in flowers and leaves is less well explored and there are still unanswered questions. These include whether plants exhibit different levels of induced responses depending on the length of exposure to the florivores. 2. To address this knowledge gap, this study used a tropical floriphilic katydid, Phaneroptera brevis, and its non‐native food plant, Lantana camara. Nursery experiments were performed in which dry matter content and anthocyanin concentration of the flower (corolla and stamens), leaf dry matter content, and leaf blade punch resistance were measured at three time phases before and after exposing the plant to the katydid individuals for 0–7 days. 3. It was demonstrated that increasing the length (days) of exposure to the katydid individual leads to higher levels of induced plant response (leaf blade punch resistance), but only in the leaves. It was also shown that higher levels of induced plant response owing to the increase in the length of exposure to the katydid individual was not observed beyond the first set of leaves developed after the exposure. 4. These results address the knowledge gap and show that plants can exhibit different levels of induced responses depending on the length of exposure to florivores. This study thus highlights the far‐reaching importance of florivory on plants.     

Influence of Plant Root Exudates on the Mobility of Fuel Volatile Compounds in Contaminated Soils

Vegetation and its associated microorganisms play an important role in the behaviour of soil contaminants. One of the most important elements is root exudation, since it can affect the mobility, and therefore, the bioavailability of soil contaminants. In this study, we evaluated the influence of root exudates on the mobility of fuel derived compounds in contaminated soils. Samples of humic acid, montmorillonite, and an A horizon from an alumi-umbric Cambisol were contaminated with volatile contaminants present in fuel: oxygenates (MTBE and ETBE) and monoaromatic compounds (benzene, toluene, ethylbenzene and xylene). Natural root exudates obtained from Holcus lanatus and Cytisus striatus and ten artificial exudates (components frequently found in natural exudates) were added to the samples, individually and as a mixture, to evaluate their effects on contaminant mobility. Fuel compounds were analyzed by headspace-gas chromatography-mass spectrometry. In general, the addition of natural and artificial exudates increased the mobility of all contaminants in humic acid. In A horizon and montmorillonite, natural or artificial exudates (as a mixture) decreased the contaminant mobility. However, artificial exudates individually had different effects: carboxylic components increased and phenolic components decreased the contaminant mobility. These results established a base for developing and improving phytoremediation processes of fuel-contaminated soils.     

Two‐tier morpho‐chemical defence tactic in Aethionema via fruit morph plasticity and glucosinolates allocation in diaspores

Fruit dimorphism and the production of glucosinolates (GSLs) are two specific life history traits found in the members of Brassicales, which aid to optimize seed dispersal and defence against antagonists, respectively. We hypothesized that the bipartite dispersal strategy demands a tight control over the production of fruit morphs with expectedly differential allocation of defensive anticipins (GSLs). In dimorphic Aethionema, herbivory by Plutella xylostella at a young stage triggered the production of more dehiscent (seeds released from fruit) than indehiscent fruit morphs (seeds enclosed within persistent pericarp) on the same plant upon maturity. Total GSL concentrations were highest in the mature seeds of dehiscent fruits from Aethionema arabicum and Aethionema saxatile among the different ontogenetic stages of the diaspores. Multivariate analyses of GSL profiles indicated significantly higher concentrations of specific indole GSLs in the diaspores, which require optimal defence after dispersal (i.e., seeds of dehiscent and fruit/pericarp of indehiscent fruit). Bioassays with a potentially coinhabitant fungus, Aspergillus quadrilineatus, support the distinct defensive potential of the diaspores corresponding to their GSL allocation. These findings indicate a two‐tier morpho‐chemical defence tactic of Aethionema via better protected fruit morphs and strategic provision of GSLs that optimize protection to the progeny for survival in nature.     

Dynamic phenotypic plasticity for root growth in Polygonum: a comparative study

Species differences in patterns of phenotypic plasticity may be an important aspect of adaptive diversity. Plasticity for functionally important root traits was studied in inbred field lineages of Polygonum persicaria and P. cespitosum (Polygonaceae). Replicate seedlings were grown in plexiglass rhizotrons under a range of constant and temporally variable moisture treatments. Plasticity was determined for final whole-plant biomass, root biomass allocation, and absolute and proportional root length. The dynamic aspect of root plasticity was examined by digitizing weekly tracings of the proportional deployment of each plant's root system to different vertical soil layers. Plants of both species expressed significant functionally adaptive phenotypic plasticity in the relative allocation, length, and vertical deployment of root systems in response to contrasting moisture conditions. Plasticity patterns in these closely related species were in general qualitatively similar, but for most traits differed in the magnitude and/or the timing of the plastic response. Dynamic changes in root deployment were more marked as well as faster in P. persicaria. Species differences in patterns of individual plasticity were generally consistent with the broader ecological distribution of P. persicaria in diverse as well as temporally variable moisture habitats.