Convergence and molecular evolution of floral fragrance after independent transitions to self-fertilization.

Studying the independent evolution of similar traits provides valuable insights into the ecological and genetic factors driving phenotypic evolution.1 The transition from outcrossing to self-fertilization is common in plant evolution2 and is often associated with a reduction in floral attractive features such as display size, chemical signals, and pollinator rewards.3 These changes are believed to result from the reallocation of the resources used for building attractive flowers, as the need to attract pollinators decreases.2,3 We investigated the similarities in the evolution of flower fragrance following independent transitions to self-fertilization in Capsella.4,5,6,7,8,9 We identified several compounds that exhibited similar changes in different selfer lineages, such that the flower scent composition reflects mating systems rather than evolutionary history within this genus. We further demonstrate that the repeated loss of ß-ocimene emission, one of the compounds most strongly affected by these transitions, was caused by mutations in different genes. In one of the Capsella selfing lineages, the loss of its emission was associated with a mutation altering subcellular localization of the ortholog of TERPENE SYNTHASE 2. This mutation appears to have been fixed early after the transition to selfing through the capture of variants segregating in the ancestral outcrossing population. The large extent of convergence in the independent evolution of flower scent, together with the evolutionary history and molecular consequences of a causal mutation, suggests that the emission of specific volatiles evolved as a response to changes in ecological pressures rather than resource limitation.

Communication between undamaged plants can elicit changes in volatile emissions from neighbouring plants, thereby altering their susceptibility to aphids.

Plant volatiles play an important role in intra- and interspecific plant communication, inducing direct and indirect defenses against insect pests. However, it remains unknown whether volatile interactions between undamaged cultivars alter host plant volatile emissions and their perception by insect pests. Here, we tested the effects of exposure of a spring barley, Hordeum vulgare L., cultivar, Salome, to volatiles from other cultivars: Fairytale and Anakin. We found that exposing Salome to Fairytale induced a significantly higher emission of trans-ß-ocimene and two unidentified compounds compared when exposed to Anakin. Aphids were repelled at a higher concentration of trans-ß-ocimene. Salome exposure to Fairytale had significant repulsive effects on aphid olfactory preference, yet not when Salome was exposed to Anakin. We demonstrate that volatile interactions between specific undamaged plants can induce changes in volatile emission by receiver plants enhancing certain compounds, which can disrupt aphid olfactory preferences. Our results highlight the significant roles of volatiles in plant-plant interactions, affecting plant-insect interactions in suppressing insect pests. This has important implications for crop protection and sustainable agriculture.

Talking Different Languages: The Role of Plant-Plant Communication When an Invader Beats up a Strange Neighborhood.

Communication through airborne volatile organic compounds (VOCs) and root exudates plays a vital role in the multifarious interactions of plants. Common ragweed (Ambrosia artemesiifolia L.) is one of the most troublesome invasive alien species in agriculture. Below- and aboveground chemical interactions of ragweed with crops might be an important factor in the invasive species' success in agriculture. In laboratory experiments, we investigated the contribution of intra- and interspecific airborne VOCs and root exudates of ragweed to its competitiveness. Wheat, soybean, and maize were exposed to VOCs emitted from ragweed and vice versa, and the adaptation response was measured through plant morphological and physiological traits. We observed significant changes in plant traits of crops in response to ragweed VOCs, characterized by lower biomass production, lower specific leaf area, or higher chlorophyll contents. After exposure to ragweed VOCs, soybean and wheat produced significantly less aboveground dry mass, whereas maize did not. Ragweed remained unaffected when exposed to VOCs from the crops or a conspecific. All crops and ragweed significantly avoided root growth toward the root exudates of ragweed. The study shows that the plant response to either above- or belowground chemical cues is highly dependent on the identity of the neighbor, pointing out the complexity of plant-plant communication in plant communities.

Alterations in the odor profile of plants in cultivar mixtures affect aphid host-location behavior.

The effect of cultivar mixtures on aphid control is attributed to the masking or alteration of host-preferred cultivar odor cues. However, the underlying physiological mechanism remains unclear. This study assessed alterations in the volatile emissions of wheat cultivars grown together (Florence-Aurora and Forment; Florence-Aurora and Montcada) and the consequences for the olfactory preference of aphids. Volatile organic compounds were collected from wheat plants grown in a laboratory under mixed or monoculture conditions and subsequently analyzed. The odor profiles of Florence-Aurora and Montcada were indistinguishable from each other. However, the odors of Florence-Aurora and Forment grown in monocultures differed significantly from those emitted by their mixture. The Florence-Aurora and Forment mixture induced plant physiological responses that affected the emission of single volatile compounds and, consequently, altered volatile organic compound ratios. English grain aphids (Sitobion avenae) were less attracted to the odors of Florence-Aurora and Forment when grown as a mixture than the combination of the odors from Florence-Aurora and Forment monocultures. Moreover, aphids preferred clean air over the odor from the Florence-Aurora and Forment mixture but preferred the odor from the Florence-Aurora and Montcada mixture over clean air. This study highlights the beneficial effects of intraspecific plant diversity on aphid control by altering plant odors in response to plant-plant interactions. The emission of less attractive odor cues consequently affects plant-aphid interactions; hence, less attractive odors are likely to impair aphid host-locating behavior. This effect was exclusive to certain cultivar mixtures, which supports the "right neighbor" concept.

Effects of Methyl Salicylate on Host Plant Acceptance and Feeding by the Aphid Rhopalosiphum padi.

Methyl salicylate (MeSA) is a volatile shown to act as an inducer of plant defense against pathogens and certain herbivores, particularly aphids. It has been shown to have potential for aphid pest management, but knowledge on its mode of action is lacking, particularly induced plant-mediated effects. This study investigated the effects of exposing plants to MeSA on the host searching, host acceptance and feeding behavior of the bird cherry-oat aphid Rhopalosiphum padi. Barley plants were exposed to volatile MeSA for 24 h, after which biological effects were tested immediately after the exposure (Day 0), and then 1, 3 and 5 days after the end of the exposure. Aphid settling on MeSA-exposed plants was significantly reduced on days 0, 1 and 3, but not on day 5. In olfactometer tests, aphids preferred the odor of unexposed plants on days 1 and 3, but not on day 0 or 5. Analysis of volatiles from exposed and unexposed plants showed higher levels of MeSA from exposed plants, most likely absorbed and re-released from plant surfaces, but also specific changes in other plant volatiles on days 0, 1 and 3. High doses of MeSA did not affect aphid orientation in an olfactometer, but lower doses were repellent. Analysis of aphid feeding by Electronic penetration graph (EPG) showed that MeSA exposure resulted in resistance factors in barley plants, including surface factors and induced systemic factors in other tissues including the phloem. The results support the potential of MeSA as a potential tool for management of aphid pests.

The effect of 1-pentadecene on Tribolium castaneum behaviour: Repellent or attractant?

BACKGROUND: Movement of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), in stored products is mediated by food volatiles and other semiochemicals. RESULTS: In two-way olfactometer assays, T. castaneum was more attracted to wheat bran previously infested with conspecifics than to uninfested bran. Chemical analysis showed that 1-pentadecene was present in the headspace of T. castaneum-infested wheat bran, but not detectable in the headspace of uninfested bran. An olfactometer was used to test the effect on T. castaneum behaviour of 1-pentadecene, and of volatiles from wheat bran with and without 1-pentadecene. The lowest concentration of 1-pentadecene exhibited an attractive effect, compared to the control (n-hexane). Slightly higher concentrations showed a neutral effect, while the highest concentrations repelled T. castaneum. Wheat bran with a low 1-pentadecene concentration was more attractive than wheat bran alone, whereas higher concentrations of 1-pentadecene were repellent. CONCLUSION: The results provide important information on intraspecific, semiochemical-mediated behaviour in T. castaneum, which could potentially be used to develop new methods to monitor the flour beetles in stored products. © 2021 Society of Chemical Industry.

Aphid feeding induces the relaxation of epigenetic control and the associated regulation of the defense response in Arabidopsis.

Environmentally induced changes in the epigenome help individuals to quickly adapt to fluctuations in the conditions of their habitats. We explored those changes in Arabidopsis thaliana plants subjected to multiple biotic and abiotic stresses, and identified transposable element (TE) activation in plants infested with the green peach aphid, Myzus persicae. We performed a genome-wide analysis mRNA expression, small RNA accumulation and DNA methylation Our results demonstrate that aphid feeding induces loss of methylation of hundreds of loci, mainly TEs. This loss of methylation has the potential to regulate gene expression and we found evidence that it is involved in the control of plant immunity genes. Accordingly, mutant plants deficient in DNA and H3K9 methylation (kyp) showed increased resistance to M. persicae infestation. Collectively, our results show that changes in DNA methylation play a significant role in the regulation of the plant transcriptional response and induction of defense response against aphid feeding.

Plant volatiles as cues and signals in plant communication.

Volatile organic compounds are important mediators of mutualistic interactions between plants and their physical and biological surroundings. Volatiles rapidly indicate competition or potential threat before these can take place, and they regulate and coordinate adaptation responses in neighbouring plants, fine-tuning them to match the exact stress encountered. Ecological specificity and context-dependency of plant-plant communication mediated by volatiles represent important factors that determine plant performance in specific environments. In this review, we synthesise the recent progress made in understanding the role of plant volatiles as mediators of plant interactions at the individual and community levels, highlighting the complexity of the plant receiver response to diverse volatile cues and signals and addressing how specific responses shape plant growth and survival. Finally, we outline the knowledge gaps and provide directions for future research. The complex dialogue between the emitter and receiver based on either volatile cues or signals determines the outcome of information exchange, which shapes the communication pattern between individuals at the community level and determines their ecological implications at other trophic levels.

Bryophytes can recognize their neighbours through volatile organic compounds.

Communication between vascular plants through volatile organic compounds (VOCs) impacts on ecosystem functioning. However, nothing is known about that between non-vascular plants. To investigate plant-plant VOCs interaction in bryophytes we exposed rare peatland moss Hamatocaulis vernicosus to VOCs of its common competitor Sphagnum flexuosum in an air-flow system of connected containers under artificial light, supplemented or unsupplemented by far-red (FR) light. When exposed to VOCs of S. flexuosum, shoots of H. vernicosus elongated and emitted six times higher amounts of a compound chemically related to ß-cyclocitral, which is employed in stress signalling and allelopathy in vascular plants. The VOCs emission was affected similarly by FR light addition, possibly simulating competition stress. This is the first evidence of plant-plant VOCs interaction in non-vascular plants, analogous to that in vascular plants. The findings open new possibilities for understanding the language and evolution of communication in land plants.

Behavioural responses of Tribolium castaneum (Herbst) to different types of uninfested and infested feed.

Tribolium castaneum is one of the most economically important insects that damages stored products. The effects of several infested or uninfested raw feed materials (wheat bran, coarse wheat meal, corn feed flour), feed products (compound feed for pigs and for laying hens) and flour mixed with brewer's yeast on the food-searching behaviour of T. castaneum adults were studied in a total of 48 combinations. Preference and olfactometer tests revealed that all the tested uninfested and intraspecific infested substrates were significantly more attractive to T. castaneum than the control (represented by part of an arena or olfactometer arm without substrate). We determined that all infested substrates were 2-9 times more attractive than uninfested in the preference test, while in the olfactometer test, they were 3-8 times more attractive. In comparing the attractiveness of the infested and uninfested substrates, in both tests wheat bran was found to be the most attractive substrate to T. castaneum adults and coarse wheat meal the least. The results of the present study contribute to our knowledge of how raw feed materials and products influence the behaviour of T. castaneum and their susceptibility to infestation, and indicate the possible utilization of wheat bran in monitoring processes of T. castaneum in pest management programmes.

Aphids-induced plant volatiles affect diel foraging behavior of a ladybird beetle Coccinella septempunctata.

The ladybird beetle Coccinella septempunctata (L.) is an important biocontrol agent of pests such as various aphid species. Despite being one of the most studied coccinellid species, many aspects of its foraging behavior are still not completely understood. This study focuses on the diel foraging behavior of C. septempunctata, investigating their olfactory orientation toward aphid-infested plants, walking activity on plants and on the soil, and feeding rates. In the scotophase the ladybird beetles were significantly more attracted to the odor of aphid-infested plants, on which they also showed considerably higher walking activity then on uninfested controls. Females were more prone to utilize olfactory cues when searching for prey and fed at higher rates than males; this shows that they are better adapted to nocturnal activity, as they require higher food intake. Coccinella septempunctata have the same feeding rate during the scotophase as in the photophase. Our study shows that C. septempunctata has the potential to forage in the scotophase if prey is abundant. The results support the hypothesis that volatiles of aphid-infested plants can attract or arrest foraging adult ladybird beetles, even in the darkness, which makes a considerable contribution to efficient prey search and enhances feeding capacity.

Who is my neighbor? Volatile cues in plant interactions.

One of the most important challenges for individual plants is coexistence with their neighbors. To compensate for their sessile lifestyle, plants developed complex and sophisticated chemical systems of communication among each other. Site-specific biotic and abiotic factors constantly alter the physiological activity of plants, which causes them to release various secondary metabolites in their environments. Volatile organic compounds (VOCs) are the most common cues that reflect a plant's current physiological status. In this sense, the identity of its immediate neighbors may have the greatest impact for a plant, as they share the same available resources. Plants constantly monitor and respond to these cues with great sensitivity and discrimination, resulting in specific changes in their growth pattern and adjusting their physiology, morphology, and phenotype accordingly. Those typical competition responses in receivers may increase their fitness as they can be elicited even before the competition takes place. Plant-plant interactions are dynamic and complex as they can include many different and important surrounding cues. A major challenge for all individual plants is detecting and actively responding only to "true" cues that point to real upcoming threat. Such selective responses to highly specific cues embedded in volatile bouquets are of great ecological importance in understanding plant-plant interactions. We have reviewed recent research on the role of VOCs in complex plant-plant interactions in plant-cross kingdom and highlighted their influence on organisms at higher trophic levels.

Disentangling olfactory and visual information used by field foraging birds.

Foraging strategies of birds can influence trophic plant-insect networks with impacts on primary plant production. Recent experiments show that some forest insectivorous birds can use herbivore-induced plant volatiles (HIPVs) to locate herbivore-infested trees, but it is unclear how birds combine or prioritize visual and olfactory information when making foraging decisions. Here, we investigated attraction of ground-foraging birds to HIPVs and visible prey in short vegetation on farmland in a series of foraging choice experiments. Birds showed an initial preference for HIPVs when visual information was the same for all choice options (i.e., one experimental setup had all options with visible prey, another setup with hidden prey). However, if the alternatives within an experimental setup included visible prey (without HIPV) in competition with HIPV-only, then birds preferred the visual option over HIPVs. Our results show that olfactory cues can play an important role in birds' foraging choices when visual information contains little variation; however, visual cues are preferred when variation is present. This suggests certain aspects of bird foraging decisions in agricultural habitats are mediated by olfactory interaction mechanisms between birds and plants. We also found that birds from variety of dietary food guilds were attracted to HIPVs; hence, the ability of birds to use plant cues is probably more general than previously thought, and may influence the biological pest control potential of birds on farmland.

Airborne signals synchronize the defenses of neighboring plants in response to touch.

Plants activate defense-related pathways in response to subtle abiotic or biotic disturbances, changing their volatile profile rapidly. How such perturbations reach and potentially affect neighboring plants is less understood. We evaluated whether brief and light touching had a cascade effect on the profile of volatiles and gene expression of the focal plant and a neighboring untouched plant. Within minutes after contact, Zea mays showed an up-regulation of certain defense genes and increased the emission of specific volatiles that primed neighboring plants, making them less attractive for aphids. Exposure to volatiles from touched plants activated many of the same defense-related genes in non-touched neighboring plants, demonstrating a transcriptional mirroring effect for expression of genes up-regulated by brief contact. Perception of so-far-overlooked touch-induced volatile organic compounds was of ecological significance as these volatiles are directly involved in plant-plant communication as an effective trigger for rapid defense synchronization among nearby plants. Our findings shed new light on mechanisms of plant responses to mechanical contact at the molecular level and on the ecological role of induced volatiles as airborne signals in plant-plant interactions.

Pest suppression in cultivar mixtures is influenced by neighbor-specific plant-plant communication.

Increased plant genotypic diversity in crop fields can promote ecosystem services including pest control, but understanding of mechanisms behind herbivore population responses to cultivar mixtures is limited. We studied aphid settling on barley plants exposed to volatiles from different cultivars, aphid population development in monocultures and two-cultivar mixtures, and differences in volatile composition between studied cultivars. Aphid responses to one cultivar in a mixture were neighbor-specific and this was more important for pest suppression than the overall mixture effect, aphid colonization patterns, or natural enemy abundance. Aphid populations decreased most in a mixture where both cultivars showed a reduced aphid-plant acceptance after reciprocal volatile exposure in the laboratory, and reduced population growth compared to monocultures in the field. Our findings suggest that herbivore population responses to crop genotypic diversity can depend on plant-plant volatile interactions, which can lead to changes in herbivore response to individual cultivars in a mixture, resulting in slower population growth. The impact of plant-plant interaction through volatiles on associated herbivore species is rarely considered, but improved understanding of these mechanisms would advance our understanding of the ecological consequences of biodiversity and guide development of sustainable agricultural practices. Combining cultivars in mixtures based on how they interact with each other is a promising strategy for sustainable pest management.

Aboveground mechanical stimuli affect belowground plant-plant communication.

Plants can detect the presence of their neighbours and modify their growth behaviour accordingly. But the extent to which this neighbour detection is mediated by abiotic stressors is not well known. In this study we tested the acclimation response of Zea mays L. seedlings through belowground interactions to the presence of their siblings exposed to brief mechano stimuli. Maize seedling simultaneously shared the growth solution of touched plants or they were transferred to the growth solution of previously touched plants. We tested the growth preferences of newly germinated seedlings toward the growth solution of touched (T_solution) or untouched plants (C_solution). The primary root of the newly germinated seedlings grew significantly less towards T_solution than to C_solution. Plants transferred to T_solution allocated more biomass to shoots and less to roots. While plants that simultaneously shared their growth solution with the touched plants produced more biomass. Results show that plant responses to neighbours can be modified by aboveground abiotic stress to those neighbours and suggest that these modifications are mediated by belowground interactions.

Plant Responses to Brief Touching: A Mechanism for Early Neighbour Detection?

In natural habitats plants can be exposed to brief and light contact with neighbouring plants. This mechanical stimulus may represent a cue that induces responses to nearby plants. However, little is known about the effect of touching on plant growth and interaction with insect herbivores. To simulate contact between plants, a soft brush was used to apply light and brief mechanical stimuli to terminal leaves of potato Solanum tuberosum L. The number of non-glandular trichomes on the leaf surface was counted on images made by light microscope while glandular trichomes and pavement cells were counted on images made under scanning electronic microscope. Volatile compounds were identified and quantified using coupled gas chromatography-mass spectrometry (GC-MS). Treated plants changed their pattern of biomass distribution; they had lower stem mass fraction and higher branch and leaf mass fraction than untouched plants. Size, weight and number of tubers were not significantly affected. Touching did not cause trichome damage nor change their total number on touched terminal leaves. However, on primary leaves the number of glandular trichomes and pavement cells was significantly increased. Touching altered the volatile emission of treated plants; they released higher quantities of the sesquiterpenes (E)-ß-caryophyllene, germacrene D-4-ol and (E)-nerolidol, and lower quantities of the terpenes (E)-ocimene and linalool, indicating a systemic effect of the treatment. The odour of touched plants was significantly less preferred by the aphids Macrosiphum euphorbiae and Myzus persicae compared to odour of untouched plants. The results suggest that light contact may have a potential role in the detection of neighbouring plants and may affect plant-insect interactions.

Red:far-red light conditions affect the emission of volatile organic compounds from barley (Hordeum vulgare), leading to altered biomass allocation in neighbouring plants.

Since the publication of this paper it has become apparent that an error was made in the scale of the vertical axis in Fig. 6I. This has no impact at all on any of the conclusions in the paper since the differences between the treatments remain as published. The authors apologise for this error and a corrected version is reproduced below.

Red:far-red light conditions affect the emission of volatile organic compounds from barley (Hordeum vulgare), leading to altered biomass allocation in neighbouring plants.

BACKGROUND AND AIMS: Volatile organic compounds (VOCs) play various roles in plant-plant interactions, and constitutively produced VOCs might act as a cue to sense neighbouring plants. Previous studies have shown that VOCs emitted from the barley (Hordeum vulgare) cultivar 'Alva' cause changes in biomass allocation in plants of the cultivar 'Kara'. Other studies have shown that shading and the low red:far-red (R:FR) conditions that prevail at high plant densities can reduce the quantity and alter the composition of the VOCs emitted by Arabidopsis thaliana, but whether this affects plant-plant signalling remains unknown. This study therefore examines the effects of far-red light enrichment on VOC emissions and plant-plant signalling between 'Alva' and 'Kara'. METHODS: The proximity of neighbouring plants was mimicked by supplemental far-red light treatment of VOC emitter plants of barley grown in growth chambers. Volatiles emitted by 'Alva' under control and far-red light-enriched conditions were analysed using gas chromatography-mass spectrometry (GC-MS). 'Kara' plants were exposed to the VOC blend emitted by the 'Alva' plants that were subjected to either of the light treatments. Dry matter partitioning, leaf area, stem and total root length were determined for 'Kara' plants exposed to 'Alva' VOCs, and also for 'Alva' plants exposed to either control or far-red-enriched light treatments. KEY RESULTS: Total VOC emissions by 'Alva' were reduced under low R:FR conditions compared with control light conditions, although individual volatile compounds were found to be either suppressed, induced or not affected by R:FR. The altered composition of the VOC blend emitted by 'Alva' plants exposed to low R:FR was found to affect carbon allocation in receiver plants of 'Kara'. CONCLUSIONS: The results indicate that changes in R:FR light conditions influence the emissions of VOCs in barley, and that these altered emissions affect VOC-mediated plant-plant interactions.

Volatile interaction between undamaged plants affects tritrophic interactions through changed plant volatile emission.

Volatile interactions between unattacked plants can lead to changes in their volatile emissions. Exposure of potato plants to onion plant volatiles results in increased emission of 2 terpenoids, (E)-nerolidol and TMTT. We investigated whether this is detectable by the ladybird Coccinella septempunctata. The odor of onion-exposed potato was significantly more attractive to ladybirds than that of unexposed potato. Further, a synthetic blend mimicking the volatile profile of onion-exposed potato was more attractive than a blend mimicking that of unexposed potato. When presented individually, TMTT was attractive to ladybirds whereas (E)-nerolidol was repellent. Volatile exchange between unattacked plants and consequent increased attractiveness for ladybirds may be a mechanism that contributes to the increased abundance of natural enemies in complex plant habitats.