Interplant Signalling Research Articles

Plant–plant communication in Camellia japonica and C. rusticana via volatiles

Plants emit volatile compounds when they are subjected to herbivorous, pathogenic, or artificial damages. Both the damaged plant and the neighboring intact plants induce resistance when they receive these volatiles, a phenomenon known as plant–plant communication. However, field observations of this phenomenon are limited. To understand the nature of plant–plant communication, we collected information about intra- and inter-plant signaling via volatiles in Camellia japonica and C. rusticana under natural conditions. We exposed intact branches of damaged plant (intra-plant) or neighboring plant (inter-plant) to artificially damaged plant volatiles (ADPVs). Leaf damage reduced in ADPVs-exposed branches in the neighboring plants compared to branches that were exposed to volatiles from intact leaves, thus, indicating that inter-plant signaling occur by the emission of volatiles from damaged leaves. We also conducted an air-transfer experiment wherein the headspace air of the damaged branch was transferred to the headspace of intact branches. Leaf damage reduced on the ADPVs-transferred branch compared to the control branch. The effect of volatiles on damage reduction lasted for three months. Our results indicate that ADPVs in Camellia species contain cues that induce resistance in neighboring plants. Our findings improve understanding of plant defense strategies that may be used in horticulture and agriculture.

Evidence of Bi-Directional Volatile-Mediated Communication between Drought-Stressed and Well-Watered Grapevines (Vitis vinifera L.)

The volatile-mediated interplay between stressed and non-stressed plants has been described in many studies involving both biotic and abiotic stresses as a one-way channel. However, very little is known about the molecular basis and mechanisms by which volatile organic compounds (VOCs) mediate plant communication between drought-stressed ‘emitter’ plants and non-stressed ‘receiver’ neighbours for the defence against impending stress challenges. Aiming to address this in grapevine, this study investigated the effect of two-way VOC exchange between stressed and non-stressed Vitis vinifera L. cv. Shiraz during drought and recovery using four treatments: isolated well-watered (WW) vines, isolated drought-stressed (DS) vines, and co-located DS ‘emitter’ and WW ‘receiver’ vines in a growth room. The results obtained from solid-phase microextraction (SPME) gas chromatography mass spectrometry (GC-MS) analysis showed a synchronised decline in α-pinene concentration in the co-located treatment vines and higher isoprene levels in the DS emitters compared to the isolated DS vines. Targeted gene expression analysis further identified the over-expression of a key gene, allene oxide synthase (AOS), in the jasmonic acid (JA) biosynthesis pathway during peak drought in the DS emitter. Transcript expression of chorismate synthase (CHORS) and α-pinene synthase (VvPNaPin1) showed similar trends in the DS emitter. The results suggest that isoprene and α-pinene may be interplant signalling molecules used by grapevine during drought. To the best of our knowledge, this is the first report of a bi-directional interaction in grapevine between the emitters and receivers under drought stress mediated by the JA and terpenoid biosynthesis pathways.

Herbivory-induced systemic signals are likely to be evolutionarily conserved in euphyllophytes.

Herbivory-induced systemic signaling has been demonstrated in monocots and dicots, and is essential for plant defense against insects. However, the nature and evolution of herbivory-induced systemic signals remain unclear. Grafting is widely used for studying systemic signaling; however, grafting between dicot plants from different families is difficult, and grafting is impossible for monocots. In this study, we took advantage of dodder's extraordinary capability of parasitizing various plant species. Field dodder (Cuscuta campestris) was employed to connect pairs of species that are phylogenetically very distant, ranging from fern to monocot and dicot plants, and so determine whether interplant signaling occurs after simulated herbivory. It was found that simulated herbivory-induced systemic signals can be transferred by dodder between a monocot and a dicot plant and even between a fern and a dicot plant, and the plants that received the systemic signals all exhibited elevated defenses. Thus, we inferred that the herbivory-induced systemic signals are likely to be evolutionarily well conserved among vascular plants. Importantly, we also demonstrate that the jasmonate pathway is probably an ancient regulator of the biosynthesis and/or transport of systemic signals in vascular plants. These findings provide new insight into the nature and evolution of systemic signaling.

Delineating induced defense responses in wheat seedlings exposed to volatiles emitted by Puccinia triticina infected wheat

Volatile organic compounds (VOCs) can play an essential role in eliciting an induced defense response in exposed plants. Preceding research showed that Puccinia triticina (Pt) infected resistant wheat emitted VOCs that induced a defense response in uninfected exposed wheat (UR) plants. The current study aimed to determine the biochemical nature of this induced defense response in the UR plants. Seedlings exposed to infected resistant plant VOCs for 8 h were used for proteomic analysis. A total of 26 proteins were up-regulated and 55 proteins were down-regulated. Five up-regulated chaperones proteins (Hsp70, chaperonin 60 subunit beta 3, ClpC, ClpC1 and ClpC2), two elongation factor-Tu, four isoforms of glyceraldehyde-3-phosphate dehydrogenase and xyloglucan galactosyltransferase are generally associated with plant defense responses. The proteomic expression profiles were supported by gene transcript expression patterns showing down-regulated phosphoglycerate kinase, up-regulated ClpC2 chaperone and unchanged carbonic anhydrase encoding transcript levels. In addition, the mitochondrial Cu/Zn superoxide dismutase (Cu/Zn-SOD Mt) encoding gene transcripts and SOD enzymatic activity were significantly induced at 8, 24 and 48 h post volatile exposure (hpe) relative to control (0 hpe). Salicylic acid quantities were significantly induced in the UR plants at 8, 24 and 48 hpe. These findings suggested that induced salicylic acid could be involved in the signaling of biochemical changes in the UR plants upon the reception of the VOCs emitted by the Pt infected wheat plants. The current study thus contributed to improving our understanding of the nature of the biochemical induced defence responses during VOCs exposed uninfected plants in an interplant signaling event.

Jasmonates: biosynthesis, perception and signal transduction.

Jasmonates (JAs) are physiologically important molecules involved in a wide range of plant responses from growth, flowering, senescence to defence against abiotic and biotic stress. They are rapidly synthesised from α-linolenic acid (ALA; C18:3 ∆9,12,15) by a process of oxidation, cyclisation and acyl chain shortening involving co-operation between the chloroplast and peroxisome. The active form of JA is the isoleucine conjugate, JA-isoleucine (JA-Ile), which is synthesised in the cytoplasm. Other active metabolites of JA include the airborne signalling molecules, methyl JA (Me-JA) and cis-jasmone (CJ), which act as inter-plant signalling molecules activating defensive genes encoding proteins and secondary compounds such as anthocyanins and alkaloids. One of the key defensive metabolites in many plants is a protease inhibitor that inactivates the protein digestive capabilities of insects, thereby, reducing their growth. The receptor for JA-Ile is a ubiquitin ligase termed as SCFCoi1 that targets the repressor protein JA Zim domain (JAZ) for degradation in the 26S proteasome. Removal of JAZ allows other transcription factors (TFs) to activate the JA response. The levels of JA-Ile are controlled through catabolism by hydroxylating enzymes of the cytochrome P450 (CYP) family. The JAZ proteins act as metabolic hubs and play key roles in cross-talk with other phytohormone signalling pathways in co-ordinating genome-wide responses. Specific subsets of JAZ proteins are involved in regulating different response outcomes such as growth inhibition versus biotic stress responses. Understanding the molecular circuits that control plant responses to pests and pathogens is a necessary pre-requisite to engineering plants with enhanced resilience to biotic challenges for improved agricultural yields.

Dodder-transmitted mobile signals prime host plants for enhanced salt tolerance

The dodders (Cuscuta spp.) are a genus of shoot parasites. In nature, a dodder often simultaneously parasitizes two or more neighboring hosts. Salt stress is a common abiotic stress for plants. It is unclear whether dodder transmits physiologically relevant salt stress-induced systemic signals among its hosts and whether these systemic signals affect the hosts' tolerance to salt stress. Here, we simultaneously parasitized two or more cucumber plants with dodder. We found that salt treatment of one host highly primed the connected host, which showed strong decreases in the extent of leaf withering and cell death in response to subsequent salt stress. Transcriptomic analysis indicated that 24 h after salt treatment of one cucumber, the transcriptome of the other dodder-connected cucumber largely resembled that of the salt-treated one, indicating that inter-plant systemic signals primed these dodder-connected cucumbers at least partly through transcriptomic reconfiguration. Furthermore, salt treatment of one of the cucumbers induced physiological changes, including altered proline contents, stomatal conductance, and photosynthetic rates, in both of the dodder-connected cucumbers. This study reveals a role of dodder in mediating salt-induced inter-plant signaling among dodder-connected hosts and highlights the physiological function of these mobile signals in plant-plant interactions under salt stress.

Stem parasitic plant Cuscuta australis (dodder) transfers herbivory-induced signals among plants

Cuscuta spp. (i.e., dodders) are stem parasites that naturally graft to their host plants to extract water and nutrients; multiple adjacent hosts are often parasitized by one or more Cuscuta plants simultaneously, forming connected plant clusters. Metabolites, proteins, and mRNAs are known to be transferred from hosts to Cuscuta, and Cuscuta bridges even facilitate host-to-host virus movement. Whether Cuscuta bridges transmit ecologically meaningful signals remains unknown. Here we show that, when host plants are connected by Cuscuta bridges, systemic herbivory signals are transmitted from attacked plants to unattacked plants, as revealed by the large transcriptomic changes in the attacked local leaves, undamaged systemic leaves of the attacked plants, and leaves of unattacked but connected hosts. The interplant signaling is largely dependent on the jasmonic acid pathway of the damaged local plants, and can be found among conspecific or heterospecific hosts of different families. Importantly, herbivore attack of one host plant elevates defensive metabolites in the other systemic Cuscuta bridge-connected hosts, resulting in enhanced resistance against insects even in several consecutively Cuscuta-connected host plants over long distances (> 100 cm). By facilitating plant-to-plant signaling, Cuscuta provides an information-based means of countering the resource-based fitness costs to their hosts.

Growth form rather than phylogenetic relationship predicts broad volatile emission patterns in the Brassicaceae

Volatile organic compounds (VOCs) released from plants are known to mediate indirect defense against herbivores and trigger intra- and interplant signaling. While systemic defense response can be mediated both via volatile and vascular signals, it is not clear whether common ancestry and/or plant growth forms influence the choice of either mode in planta. We hypothesize that larger woody plants with a complex anatomy should rely more on volatile-mediated signaling, apparently to circumvent vascular restrictions that slow down the communication over a large distance. On the other hand, in smaller herbaceous plants faster systemic response can be achieved via vascular signaling. To investigate whether plant VOCs emission is related to plant phylogeny or growth form, we studied the composition of herbivory-induced plant volatiles in 13 Brassicaceae species representing all four evolutionary lineages, because this family is characterized by both a well-resolved phylogeny and highly diverse growth forms. Our results revealed that woody species consistently emitted a more complex blend of volatiles than herbaceous species. However, phylogenetic relatedness of the species did not explain the observed volatile emission patterns. This emphasizes the influence of growth form, rather than phylogenetic relationships on the variation in plant volatile emissions. Our findings suggest that woody, perennial plant species emit diverse VOCs, likely because these compounds comprise a more efficient mode of defense response in these large, anatomically complex plants.

Terpenoids in plant and arbuscular mycorrhiza-reinforced defence against herbivorous insects.

Plants, though sessile, employ various strategies to defend themselves against herbivorous insects and convey signals of an impending herbivore attack to other plant(s). Strategies include the production of volatiles that include terpenoids and the formation of symbiotic associations with fungi, such as arbuscular mycorrhiza (AM). This constitutes a two-pronged above-ground/below-ground attack-defence strategy against insect herbivores. Terpenoids represent an important constituent of herbivore-induced plant volatiles that deter herbivores and/or attract their predators. Terpenoids serve as airborne signals that can induce defence responses in systemic undamaged parts of the plant and also prime defence responses in neighbouring plants. Colonization of roots by AM fungi is known to influence secondary metabolism in plants; this includes alteration of the concentration and composition of terpenoids, which can boost both direct and indirect plant defence against herbivorous insects. Enhanced nutrient uptake facilitated by AM, changes in plant morphology and physiology and increased transcription levels of certain genes involved in the terpenoid biosynthesis pathway result in alterations in plant terpenoid profiles. The common mycorrhizal networks of external hyphae have added a dimension to the two-pronged plant defence strategy. These act as conduits to transfer defence signals and terpenoids. Improved understanding of the roles of terpenoids in plant and AM defences against herbivory and of interplant signalling in natural communities has significant implications for sustainable management of pests in agricultural ecosystems.

Root-emitted volatile organic compounds: can they mediate belowground plant-plant interactions?

Aboveground, plants release volatile organic compounds (VOCs) that act as chemical signals between neighbouring plants. It is now well documented that VOCs emitted by the roots in the plant rhizosphere also play important ecological roles in the soil ecosystem, notably in plant defence because they are involved in interactions between plants, phytophagous pests and organisms of the third trophic level. The roles played by root-emitted VOCs in between- and within-plant signalling, however, are still poorly documented in the scientific literature. Given that (1) plants release volatile cues mediating plant-plant interactions aboveground, (2) roots can detect the chemical signals originating from their neighbours, and (3) roots release VOCs involved in biotic interactions belowground, the aim of this paper is to discuss the roles of VOCs in between- and within-plant signalling belowground. We also highlight the technical challenges associated with the analysis of root-emitted VOCs and the design of experiments targeting volatile-mediated root-root interactions. We conclude that root-root interactions mediated by volatile cues deserve more research attention and that both the analytical tools and methods developed to study the ecological roles played by VOCs in interplant signalling aboveground can be adapted to focus on the roles played by root-emitted VOCs in between- and within-plant signalling.

Volatiles emitted by leaf rust infected wheat induce a defence response in exposed uninfected wheat seedlings

Volatile organic compounds emitted from plant tissues have been implicated to play many roles. Interplant signalling is an experimentally proven phenomenon whereby volatiles produced under stress conditions may diffuse to neighbouring plants to induce a response. The current project was undertaken to investigate a putative signalling event between leaf rust (Puccinia triticina Erikss.) infected and uninfected wheat (Triticum aestivum L.) using a continual air-flow system. Exposure to volatiles emitted by either infected resistant or susceptible plants decreased the percentage leaf area infected and pustule size in recipient plants after a subsequent infection with leaf rust, but in the case of infected resistant plants, the decrease in percentage leaf area infected was significant. The activation of a defence response was confirmed through a significant induction of β-1,3-glucanase activity at 8 h post volatile exposure in exposed resistant and susceptible seedlings, depending on the source of the volatiles. Increased PR1 gene expression supported the activation of the defence response. Emitted volatiles were captured by solid phase micro-extraction and identified by GC-MS. Based on their detection and the literature, ocimene from infected resistant seedlings and green leaf volatiles from infected susceptible seedlings are suggested as likely candidates for the inter-plant communication.

Interplant signalling through hyphal networks.

Mycorrhizal fungi can form common mycelial networks (CMNs) that interconnect plants. Here, we provide an insight into recent findings demonstrating that CMNs can be conduits for interplant signalling, influencing defence against insect herbivores and foliar necrotrophic fungi. A likely mechanism is direct transfer of signalling molecules within hyphae. However, electrical signals, which can be induced by wounding, may also enable signalling over relatively long distances, because the biophysical constraints imposed by liquid transport in hyphae and interaction with soil are relieved. We do not yet understand the ecological, evolutionary and agronomic implications of interplant signalling via CMNs. Identifying the mechanism of interplant signalling will help to address these gaps.

A role for volatiles in intra- and inter-plant interactions in birch.

One of the first observations that plants might utilize cues released by damaged neighbors under natural conditions was made in birch forests in 1985. However, the mechanisms underlying the observations were not determined, and birch (Betula spp.) has been neglected as a study system for inter-plant interaction ever since. Volatiles released by vegetative plant parts in response to herbivore damage play important roles as signals in plant-to-plant interactions in a range of woody and herbaceous plant species, and also have been shown to mediate signaling between branches of the same plant that have limited vascular connection. We established greenhouse experiments to assess: 1) whether exposure to plant volatiles from herbivore-damaged birches primes defense responses in undamaged neighbors; and 2) whether defenses also are primed in undamaged parts of the same plants with limited vascular connection. We observed a priming of defense responses, which were manifested in an augmented emission of terpenes and aromatic compounds in undamaged conspecific neighbors, and also an augmented emission of green leaf volatiles in systemic branches. Our work provides strong evidence of inter-plant signaling by volatiles, and an intra-plant systemic response in birch. However, the responses are specific, with emissions of different groups of plant volatiles typifying the primed response. This work complements and extends the previous work conducted with a natural population of birches.

Leaf Mimicry: Chameleon-like Leaves in a Patagonian Vine

Mimicry has evolved in plants for a number of traits, both floral and vegetative. The discovery of a vine that mimics the leaf shape of different hosts poses new questions about the function of leaf mimicry, interplant signalling and leaf development.

Interplant volatile signaling in willows: revisiting the original talking trees

The importance of interplant volatile signaling in plant-herbivore interactions has been a contentious issue for the past 30 years. We revisit willows as the system in which evidence for interplant signaling was originally found, but then questioned. We established three well-replicated experiments with two willow species (Salix exigua and Salix lemmonii) to address whether the receipt of an interplant signal from a neighboring willow reduces herbivore damage. Additionally we tested whether this signal is volatile in nature, and whether plants signal better to themselves than they do to other individuals. In all three experiments, we found evidence that cues from a damaged neighbor reduce subsequent herbivory experienced by willows. In one experiment, we showed that bagging of clipped tissue, which prevents the exchange of volatile signals, removed the effect of neighbor wounding. This was consistent with results from the other two experiments, in which clipping potted neighbors connected only through airborne volatile cues reduced damage of receivers. In one year, we found evidence that the perception of volatile signals from genetically identical clones was more effective at reducing foliar damage to a neighbor than signals from a genetically different individual. However, this trend was not significant in the following year. In three well-replicated experiments, we found strong evidence for the importance of interplant volatile cues in mediating herbivore interactions with willows.

Four terpene synthases produce major compounds of the gypsy moth feeding-induced volatile blend of Populus trichocarpa

After herbivore damage, many plants increase their emission of volatile compounds, with terpenes usually comprising the major group of induced volatiles. Populus trichocarpa is the first woody species with a fully sequenced genome, enabling rapid molecular approaches towards characterization of volatile terpene biosynthesis in this and other poplar species. We identified and characterized four terpene synthases (PtTPS1–4) from P. trichocarpa which form major terpene compounds of the volatile blend induced by gypsy moth ( Lymantria dispar) feeding. The enzymes were heterologously expressed and assayed with potential prenyl diphosphate substrates. PtTPS1 and PtTPS2 accepted only farnesyl diphosphate and produced (−)-germacrene D and ( E, E)-α-farnesene as their major products, respectively. In contrast, PtTPS3 and PtTPS4 showed both mono- and sesquiterpene synthase activity. They produce the acyclic terpene alcohols linalool and nerolidol but exhibited opposite stereospecificity. qRT-PCR analysis revealed that the expression of the respective terpene synthase genes was induced after feeding of gypsy moth caterpillars. The TPS enzyme products may play important roles in indirect defense of poplar to herbivores and in mediating intra- and inter-plant signaling.

Volatiles of myrmecophytic Piper plants signal stem tissue damage to inhabiting Pheidole ant‐partners

Summary The major aim of this paper was to explore the mechanistic basis of induced defensive ant response in myrmecophytic Piper species and their ant‐partner Pheidole bicornis by combining experimental trials and chemical analysis. In field experiments, the response of Pheidole ants to simulated stem‐damage of their host plant was documented. Their reaction to other myrmecophytic and non‐myrmecophytic Piper species was explored by fixing respective stem sections onto inhabited Piper fimbriulatum individuals. After artificial damage to stems of P. fimbriulatum and stem sections from other myrmecophytic Piper, the number of ants increased to a maximum of 63 individuals on the damaged internode within the first 10 min. By contrast, no or only a poor reaction was elicited by stem sections from the selected non‐myrmecophytic Piper species. Solid‐phase microextraction/gas chromatography–mass spectrometry identified 68 volatile organic compounds (VOCs) which were emitted from wounded stem bark of three myrmecophytic (P. fimbriulatum, P. obliquum, P. sagittifolium) and two non‐myrmecophytic (P. hispidum, P. riparense) Piper species, comprising monoterpenes, sesquiterpenes, simple aromatic compounds, and green leaf volatiles (GLVs, oxylipins). Despite considerable variation in the VOC profiles, an analysis of similarities (anosim) classified the accumulation of the sesquiterpenes β‐caryophyllene, germacrene D and emission of the oxylipin reactive electrophilic species (RES) compounds hexanal, cis‐3‐hexene‐1‐ol, 2‐hexenal, 2‐undecanone and 1‐hexanol as characteristic of the myrmecophytic species. Synthesis. Field experiments showed that mechanical disturbance of myrmecophytic Piper stems is not sufficient to elicit recruitment and alertness of the ant‐partner Ph. bicornis, but that specific volatiles released from the host plant are responsible. Analyses of the VOCs identified after simulated stem damage of three myrmecophytic and two non‐myrmecophytic Piper species revealed differences in their VOC emission profiles: Only in the myrmecophytic species were induced stress compounds found, such as various oxylipin RES or the sesquiterpene β‐caryophyllene, which are typically emitted after plant damage for interplant signalling. Some of these compounds (e.g. hexanal, 1‐hexanol) are known to induce alerting and alarm behaviours in several ant species. Our study provides evidence that the VOCs of the Piper host plant may signal stem damage and induce recruitment and alert behaviour of associated Pheidole ants.

Plant–Plant Signaling: Application of trans- or cis-Methyl Jasmonate Equivalent to Sagebrush Releases Does Not Elicit Direct Defenses in Native Tobacco

Nicotiana attenuata plants growing in close proximity to damaged sagebrush (Artemisia tridentata ssp. tridentata) suffer less herbivory than plants near undamaged sagebrush. Sagebrush constitutively releases methyl jasmonate (MeJA), a compound that when applied directly to N. attenuata, elicits herbivore resistance and the direct defense traits [protease inhibitors (PIs), nicotine]. Damage increases the release of volatile MeJA, primarily in the cis epimer, suggesting that cis-MeJA may mediate this apparent interplant signaling. We characterized sagebrush's MeJA plume before and after damage in nature and in the laboratory, and compared the activity of trans- and cis-MeJA in inducing PIs, nicotine, and Manduca sexta resistance in N. attenuata. We used both lanolin applications and aqueous sprays that mimic natural exposures, and we determined the amount of volatilized MeJA required to elicit a nicotine response in open-grown plants. Wounding rapidly and transiently increased cis-MeJA emissions from damaged parts (but not systemically), and the released plume did not rapidly dissipate in nature. cis-MeJA was not consistently more active than trans-MeJA, and the order of exposure (trans- then cis-) did not influence activity. We conclude that volatile MeJA, either trans- or cis-, when applied at levels consistent with those released by sagebrush does not elicit direct defenses in N. attenuata.

Chemical information transfer between wounded and unwounded plants: backing up the future

Abstract This special issue on “Chemical information transfer between wounded and unwounded plants” provides an overview of past and ongoing experiments on plant-to-plant communication. Since the studies on plant responses to single gaseous compounds were not particularly emphasised, the actual number of studies relevant to the subject is underestimated. All in all, we think the amount of data on damage-induced plant-to-plant information transfer makes that the phenomenon can no longer be denied and deserves intensified attention by the scientific community. In this epilogue we highlight a couple of issues which received little attention and present some speculative ideas. First we concentrate on functional aspects of plant-plant communication we stress the concept of damage-induced signalling as an ecological cost to the signal-sending plant and we discuss the theoretical development on interplant signalling, which is still in its infancy. With respect to mechanisms, we compare above- to belowground signalling, discuss potential cues and stress the possibility that responses in signal-exposed plants may be hidden. Finally, we address some future prospects which may help in the further development of the still underexposed phenomenon of damage-induced plant-to-plant information transfer.