Mycorrhiza-induced resistance in citrus against Tetranychus urticae is plant species dependent and inversely correlated to basal immunity.

BACKGROUND: Mycorrhizal plants show enhanced resistance to biotic stresses, but few studies have addressed mycorrhiza-induced resistance (MIR) against biotic challenges in woody plants, particularly citrus. Here we present a comparative study of two citrus species, Citrus aurantium, which is resistant to Tetranychus urticae, and Citrus reshni, which is highly susceptible to T. urticae. Although both mycorrhizal species are protected in locally infested leaves, they show very distinct responses to MIR. RESULTS: Previous studies have indicated that C. aurantium is insensitive to MIR in systemic tissues and MIR-triggered antixenosis. Conversely, C. reshni is highly responsive to MIR which triggers local, systemic and indirect defense, and antixenosis against the pest. Transcriptional, hormonal and inhibition assays in C. reshni indicated the regulation of jasmonic acid (JA)- and abscisic acid-dependent responses in MIR. The phytohormone jasmonic acid isoleucine (JA-Ile) and the JA biosynthesis gene LOX2 are primed at early timepoints. Evidence indicates a metabolic flux from phenylpropanoids to specific flavones that are primed at 24 h post infestation (hpi). MIR also triggers the priming of naringenin in mycorrhizal C. reshni, which shows a strong correlation with several flavones and JA-Ile that over-accumulate in mycorrhizal plants. Treatment with an inhibitor of phenylpropanoid biosynthesis C4H enzyme impaired resistance and reduced the symbiosis, demonstrating that phenylpropanoids and derivatives mediate MIR in C. reshni. CONCLUSION: MIR's effectiveness is inversely correlated to basal immunity in different citrus species, and provides multifaceted protection against T. urticae in susceptible C. reshni, activating rapid local and systemic defenses that are mainly regulated by the accumulation of specific flavones and priming of JA-dependent responses. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Identification of potential invasive alien species in Spain through horizon scanning.

Invasive alien species have widespread impacts on native biodiversity and ecosystem services. Since the number of introductions worldwide is continuously rising, it is essential to prevent the entry, establishment and spread of new alien species through a systematic examination of future potential threats. Applying a three-step horizon scanning consensus method, we evaluated non-established alien species that could potentially arrive, establish and cause major ecological impact in Spain within the next 10 years. Overall, we identified 47 species with a very high risk (e.g. Oreochromis niloticus, Popillia japonica, Hemidactylus frenatus, Crassula helmsii or Halophila stipulacea), 61 with high risk, 93 with moderate risk, and 732 species with low risk. Many of the species categorized as very high or high risk to Spanish biodiversity are either already present in Europe and neighbouring countries or have a long invasive history elsewhere. This study provides an updated list of potential invasive alien species useful for prioritizing efforts and resources against their introduction. Compared to previous horizon scanning exercises in Spain, the current study screens potential invaders from a wider range of terrestrial, freshwater, and marine organisms, and can serve as a basis for more comprehensive risk analyses to improve management and increase the efficiency of the early warning and rapid response framework for invasive alien species. We also stress the usefulness of measuring agreement and consistency as two different properties of the reliability of expert scores, in order to more easily elaborate consensus ranked lists of potential invasive alien species.

Mycorrhizal Symbiosis Triggers Local Resistance in Citrus Plants Against Spider Mites.

Citrus plants are a highly mycotrophic species with high levels of fungal colonization. Citrus aurantium rootstocks typically show abundant root colonization by Rhizophagus irregularis three weeks after inoculation. Mycorrhizal symbiosis protects plants against multiple biotic stressors, however, such protection against spider mites remains controversial. We examined mycorrhiza-induced resistance (MIR) in citrus against the two-spotted spider mite Tetranychus urticae. Mycorrhized C. aurantium displayed reduced levels of damage in leaves and lower mite oviposition rates, compared to non-mycorrhized controls. Mycorrhization did not affect host choice of mites in Y-tube assays; of note, C. aurantium has innate strong antixenotic resistance against this mite. Analysis of metabolism pathways in mycorrhized citrus plants showed upregulated expression of the oxylipin-related genes LOX-2 and PR-3 early after infestation. Accordingly, jasmonic acid (JA), 12-oxo phytodienoic acid (OPDA), and JA-Ile concentrations were increased by mycorrhization. Non-targeted metabolomic analysis revealed the amino acid, oxocarboxylic acid, and phenylpropanoid metabolism as the three major pathways with more hits at 24 h post infection (hpi) in mycorrhized plants. Interestingly, there was a transition to a priming profile of these pathways at 48 hpi following infestation. Three flavonoids (i.e., malic acid, coumaric acid, and diconiferyl alcohol) were among the priming compounds. A mixture containing all these compounds provided efficient protection against the mite. Unexpectedly, systemic resistance did not improve after 72 h of primary infestation, probably due to the innate strong systemic resistance of C. aurantium. This is the first study to show that MIR is functional against T. urticae in locally infested citrus leaves, which is mediated by a complex pool of secondary metabolites and is likely coordinated by priming of JA-dependent responses.

Opposing roles of plant laticifer cells in the resistance to insect herbivores and fungal pathogens.

More than 12,000 plant species (ca. 10% of flowering plants) exude latex when their tissues are injured. Latex is produced and stored in specialized cells named "laticifers". Laticifers form a tubing system composed of rows of elongated cells that branch and create an internal network encompassing the entire plant. Laticifers constitute a recent evolutionary achievement in ecophysiological adaptation to specific natural environments; however, their fitness benefit to the plant still remains to be proven. The identification of Euphorbia lathyris mutants (pil mutants) deficient in laticifer cells or latex metabolism, and therefore compromised in latex production, allowed us to test the importance of laticifers in pest resistance. We provided genetic evidence indicating that laticifers represent a cellular adaptation for an essential defense strategy to fend off arthropod herbivores with different feeding habits, such as Spodoptera exigua and Tetranychus urticae. In marked contrast, we also discovered that a lack of laticifer cells causes complete resistance to the fungal pathogen Botrytis cinerea. Thereafter, a latex-derived factor required for conidia germination on the leaf surface was identified. This factor promoted disease susceptibility enhancement even in the non-latex-bearing plant Arabidopsis. We speculate on the role of laticifers in the co-evolutionary arms race between plants and their enemies.

Plant-feeding may explain why the generalist predator Euseius stipulatus does better on less defended citrus plants but Tetranychus-specialists Neoseiulus californicus and Phytoseiulus persimilis do not.

The generalist predator Euseius stipulatus (Athias-Henriot) and the Tetranychidae-specialist predators Neoseiulus californicus (McGregor) and Phytoseiulus persimilis Athias-Henriot play a key role in the regulation of Tetranychus urticae Koch in Spanish citrus orchards. Previous studies have shown that sour orange (Citrus aurantium L.) and Cleopatra mandarin (Citrus reshni hort. ex Tan.) display extreme resistance and susceptibility to T. urticae, respectively. When offered a choice between these two genotypes infested by T. urticae, E. stipulatus preferred Cleopatra mandarin, whereas the specialists did not show any preference. The present study was undertaken to check whether these preferences could be related to the feeding of E. stipulatus on the host plant and/or to differences in prey feeding on the two plants. Our results demonstrate that E. stipulatus is a zoophytophagous mite, which can engage in direct plant feeding in sour orange and Cleopatra mandarin, whereas neither N. californicus nor P. persimilis do so. Whereas Cleopatra mandarin provided a higher-quality prey/feeding substrate for E. stipulatus, which may be related to its phytophagy, no differences were observed for the two specialists. As higher constitutive and faster inducible defense against T. urticae in sour orange relative to Cleopatra mandarin plants result in sour orange supporting lower T. urticae densities and plant damage, our results demonstrate that pest regulation by specialist natural enemies may be more effective when prey feed on better defended plants.

Practices to Conserve Pollinators and Natural Enemies in Agro-Ecosystems.

Intensive agriculture has put great pressure on populations of beneficial arthropods such as natural enemies and pollinators, especially through adverse effects of pesticide use and the impact on resources in the agricultural landscape, i [...].

Molecular characterization of Cardinium, Rickettsia, Spiroplasma and Wolbachia in mite species from citrus orchards.

Tetranychidae spider mites are considered key citrus pests in some production areas, especially Tetranychus urticae Koch. Over the past decades, pesticide overuse seems to have promoted T. urticae population selection in citrus orchards. However, the microbiota has also been pointed out as a plausible explanation for population structure or plant host specialisation observed in several arthropod species. In this work, we have determined the incidence of Cardinium, Rickettsia, Spiroplasma and Wolbachia as representatives of major distorter bacteria genera in Aplonobia histricina (Berlese), Eutetranychus banksi (McGregor), Eutetranychus orientalis (Klein), Panonychus citri (McGregor), Tetranychus evansi Baker and Pritchard, Tetranychus turkestani Ugarov and Nikolskii, and T. urticae populations from Spanish citrus orchards. Only Wolbachia was detected by PCR. The multilocus alignment approach and phylogenetic inference indicated that all detected Wolbachia belong to supergroup B. The deep analysis of each 16S rDNA, ftsZ and wsp gene sequences allowed identifying several phylogenetically different Wolbachia sequences. It probably indicates the presence of several different races or strains, all of them belonging to supergroup B. The wsp sequence typing analysis unveiled the presence of the two already identified alleles (61 and 370) and allowed to contribute with five new alleles, supporting the presence of different but related B-races in the studied mite populations. The results are discussed and related to T. urticae population structure, previously observed in Spanish citrus orchards.

Tracking mite trophic interactions by multiplex PCR.

BACKGROUND: A thorough knowledge of trophic webs in agroecosystems is essential to achieve successful biological pest control. Phytoseiid mites are the most efficient natural enemies of tetranychid mites, which include several important pests worldwide. Nevertheless, phytoseiids may feed on other food sources including other microarthropods, plants and even other phytoseiids (intraguild predation), which can interfere with biological control services. Molecular gut content analysis is a valuable tool for characterizing trophic interactions, mainly when working on microarthropods such as mites. We have designed new primers for Phytoseiidae, Tetranychidae and Thysanoptera identification and they have been multiplexed in a polymerase chain reaction (PCR) together with universal plant primers. Additionally, we have estimated prey DNA detectability success over time (DS50 ) considering the most probable events in Spanish citrus orchards: the phytoseiid Euseius stipulatus as a predator, the phytoseiid Phytoseiulus persimilis as intraguild prey, and the thrips Frankliniella occidentalis and Anaphothrips obscurus as alternative prey to Tetranychus urticae. RESULTS: The designed multiplex PCR allows the identification of phytoseiids (both predator and intraguild prey) and detects alternative food sources mentioned above in the gut of the phytoseiid predator. DS50 for E. stipulatus as the predator were 1.3, 2.3 and 18.7 h post feeding for F. occidentalis, A. obscurus and P. persimilis as prey, respectively. CONCLUSION: Tracking of the trophic relationships within the citrus acarofauna, and the unveiling of the role of alternative food sources will pave the way for enhancing T. urticae biological control. This multiplex PCR approach could be applicable for these purposes in similar agroecosystems. © 2019 Society of Chemical Industry.

Zoophytophagous mites can trigger plant-genotype specific defensive responses affecting potential prey beyond predation: the case of Euseius stipulatus and Tetranychus urticae in citrus.

BACKGROUND: Zoophytophagous predators can trigger plant defenses affecting prey populations beyond predation. Euseius stipulatus is a presumed zoophytophagous phytoseiid common in citrus. The response of citrus to one of its potential prey, Tetranychus urticae, is genotype dependent, with Citrus reshni and C. aurantium exhibiting extreme susceptibility and resistance, respectively. Volatile blends produced upon infestation affected the behavior of these two mites. We wondered whether E. stipulatus could trigger similar responses. RESULTS: Euseius stipulatus triggered genotype-dependent defense responses in citrus. Whereas C. aurantium upregulated the Jasmonic Acid, Salicylic Acid and flavonoids defensive pathways, C. reshni upregulated JA only. Likewise, different volatile blends were induced. These blends were exploited by E. stipulatus to select less-defended plants (i.e., those in which higher pest densities are expected) and, interestingly, did not prevent T. urticae from choosing E. stipulatus-infested plants. To the best of our knowledge, this is the first time that this type of response has been described for a zoophytophagous phytoseiid. CONCLUSION: The observed responses could affect herbivore populations through plant-mediated effects. Although further research is needed to fully characterize them and include other arthropods in the system, these results open opportunities for more sustainable and effective pest control methods (i.e., combining semiochemicals and biological control). © 2018 Society of Chemical Industry.

Can Plant Defence Mechanisms Provide New Approaches for the Sustainable Control of the Two-Spotted Spider Mite Tetranychus urticae?

Tetranychus urticae (T. urticae) Koch is a cosmopolitan, polyphagous mite which causes economic losses in both agricultural and ornamental plants. Some traits of T. urticae hamper its management, including a short life cycle, arrhenotokous parthenogenesis, its haplodiploid sex determination system, and its extraordinary ability to adapt to different hosts and environmental conditions. Currently, the use of chemical and biological control are the major control methods used against this mite. In recent years, some studies have focused on plant defence mechanisms against herbivores. Various families of plant compounds (such as flavonoids, glucosinolates, or acyl sugars) have been shown to behave as acaricides. Plants can be induced upon appropriate stimuli to increase their resistance against spider mites. This knowledge, together with the understanding of mechanisms by which T. urticae detoxifies and adapts to pesticides, may complement the control of this pest. Herein, we describe plant volatile compounds (VOCs) with repellent activity, and new findings about defence priming against spider mites, which interfere with the T. urticae performance. The use of VOCs and defence priming can be integrated into current management practices and reduce the damage caused by T. urticae in the field by implementing new, more sustainable crop management tools.

Development of an attract-and-infect system to control Rhynchophorus ferrugineus with the entomopathogenic fungus Beauveria bassiana.

BACKGROUND: A new Beauveria bassiana-based attract and infect device (AID) to control Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) was developed. The virulence and persistence of the fungal formulation used in the AID were evaluated in the laboratory. Semi-field and field trials were carried out to validate the results and establish the potential of this device as a control tool. RESULTS: In laboratory conditions, a 50% lethal time (LT50 ) of 4.33 days was obtained when adults (7-10 days old) were exposed to the inoculation tunnel (IT) containing 1 × 1010 conidia g-1 in an oil-based fungal formulation. This formulation maintained conidium viability at 50% for up to 2 months. Moreover, when adults were exposed to 2.5-month field-aged ITs, mortality still reached 50% 40 days after exposure. In addition, no differences were observed between ITs aged in early spring and those aged in summer, suggesting that the fungal formulation is not strongly affected by environmental factors in Mediterranean basin conditions. Semi-field assays showed that the device allowed an easy transit of weevils through the IT, which were effectively attracted and infected. Using the AIDs in 4-ha plot field trials, a reduction of >50% in the percentage of infested sentinel palms was obtained. CONCLUSION: Based on the results obtained in terms of the efficacy and persistence of this new AID in the field and its potential in reducing R. ferrugineus populations and palm infestation, this device could become a key tool for the management of R. ferrugineus. © 2018 Society of Chemical Industry.

When do predatory mites (Phytoseiidae) attack? Understanding their diel and seasonal predation patterns.

Predatory mites of the Phytoseiidae family are considered one of the most important groups of natural enemies used in biological control. The behavioral patterns of arthropods can differ greatly daily and seasonally; however, there is a lack of literature related to Phytoseiidae diel and seasonal predation patterns. The predatory activity of three phytoseiid species (two Tetranychidae-specialists, Phytoseiulus persimilis and Neoseiulus californicus, and one omnivore, Euseius stipulatus) that occur naturally in Spanish citrus orchards was observed under laboratory conditions in winter and summer. The temperature and photoperiod of the climatic chamber where the mites were reared did not change during the experiment. Our study demonstrates that phytoseiids can exhibit diel and seasonal predatory patterns when feeding on Tetranychus urticae (Acari: Tetranychidae). Neoseiulus californicus was revealed to be a nocturnal predator in summer but diurnal in winter. In contrast, P. persimilis activity was maximal during the daytime, and E. stipulatus showed no clear daily predation patterns. The predatory patterns described in this study should be taken into account when designing laboratory studies and also in field samplings, especially when applying molecular techniques to unveil trophic relationships.

Zoophytophagous mirids provide pest control by inducing direct defences, antixenosis and attraction to parasitoids in sweet pepper plants.

BACKGROUND: In addition to their services as predators, mirid predators are able to induce plant defences by phytophagy. However, whether this induction occurs in sweet pepper and whether it could be an additional benefit to their role as a biological control agent in this crop remain unknown. Here, these questions were investigated in two model insects, the mirids Nesidiocoris tenuis and Macrolophus pygmaeus. RESULTS: Plant feeding behaviour was observed in both N. tenuis and M. pygmaeus on sweet pepper and occupied 33% and 14% of total time spent on the plant, respectively. The punctures caused by mirid plant feeding induced the release of a blend of volatile organic compounds (VOCs) which repelled the herbivore pests Frankliniella occidentalis and Bemisia tabaci and attracted the whitefly parasitoid Encarsia formosa. The repellent effect on B. tabaci was observed for at least 7 days after initial exposure of the plant to N. tenuis, and attraction of E. formosa remained functional for 14 days. CONCLUSION: Plant defences induced by the feeding of mirid predators, their subsequent effects on the behaviour of both pests and natural enemies, and the persistence of these observed effects open the door to new control strategies in the sweet pepper crop. Further application of this research is discussed, such as the vaccination of plants by zoophytophagous mirids in the nursery before transplantation. © 2017 Society of Chemical Industry.

Mite diversity (Acari: Tetranychidae, Tydeidae, Iolinidae, Phytoseiidae) and within-tree distribution in citrus orchards in southern Spain, with special reference to Eutetranychus orientalis.

Mite diversity of selected citrus orchards in Andalusia (southern Spain) was studied during 2002-2007. The following species were found: Eutetranychus orientalis, Tetranychus urticae, Panonychus citri (Tetranychidae), Tydeus californicus, Lorryia formosa (Tydeidae), Pronematus ubiquitus (Iolinidae), Euseius stipulatus, Typhlodromus phialatus, Neoseiulus californicus, Euseius scutalis, Phytoseiulus persimilis, Paraseiulus talbii and Neoseiulus cucumeris (Phytoseiidae). Eutetranychus orientalis was the dominant tetranychid species in orange and lemon trees, whereas T. urticae was slightly more abundant in mandarines. The most abundant tydeid in clementine and orange was Tydeus californicus, in lemon it was L. formosa. The iolinid P. ubiquitus was found in very low numbers. With respect to phytoseiids, E. stipulatus was dominant in the three citrus species. Eutetranychus orientalis had a unimodal phenology, peaking only in autumn, whereas the other two tetranychids had two maxima. Tydeus californicus showed one or two peaks depending upon the citrus species, and E. stipulatus also had two peaks, one in spring and one in autumn. The mites displayed a non-random distribution in the tree. Eutetranychus orientalis preferred the outer and upper leaves. On the contrary, tydeids (mainly T. californicus) and phytoseiids (mainly E. stipulatus) preferred inner and lower leaves. These preferences had not been sufficiently described previously, and they are important for sampling plans.

Systemic resistance in citrus to Tetranychus urticae induced by conspecifics is transmitted by grafting and mediated by mobile amino acids.

Recent research suggests that systemic signalling and communication between roots and leaves plays an important role in plant defence against herbivores. In the present study, we show that the oviposition of the two-spotted spider mite Tetranychus urticae in the systemic leaves of citrus rootstock Citrus aurantium (sour orange) was reduced by 50% when a lower leaf was previously infested with conspecifics. Metabolomic and gene expression analysis of the root efflux revealed a strong accumulation of glutamic acid (Glu) that triggered the expression of the citrus putative glutamate receptor (GRL) in the shoots. Additionally, uninfested sour orange systemic leaves showed increased expression of glutamate receptors and higher amounts of jasmonic acid (JA) and 12-oxo-phytodienoic acid in plants that were previously infested. Glu perception in the shoots induced the JA pathway, which primed LOX-2 gene expression when citrus plants were exposed to a second infestation. The spider mite-susceptible citrus rootstock Cleopatra mandarin (C. unshiu) also expressed systemic resistance, although the resistance was less effective than the resistance in sour orange. Surprisingly, the mobile signal in Cleopatra mandarin was not Glu, which suggests a strong genotype-dependency for systemic signalling in citrus. When the cultivar Clemenules (C. clementina) was grafted onto sour orange, there was a reduction in symptomatic leaves and T. urticae populations compared to the same cultivar grafted onto Cleopatra mandarin. Thus, systemic resistance is transmitted from the roots to the shoots in citrus and is dependent on rootstock resistance.

Stage-Related Defense Response Induction in Tomato Plants by Nesidiocoris tenuis.

The beneficial effects of direct predation by zoophytophagous biological control agents (BCAs), such as the mirid bug Nesidiocoris tenuis, are well-known. However, the benefits of zoophytophagous BCAs' relation with host plants, via induction of plant defensive responses, have not been investigated until recently. To date, only the females of certain zoophytophagous BCAs have been demonstrated to induce defensive plant responses in tomato plants. The aim of this work was to determine whether nymphs, adult females, and adult males of N. tenuis are able to induce defense responses in tomato plants. Compared to undamaged tomato plants (i.e., not exposed to the mirid), plants on which young or mature nymphs, or adult males or females of N. tenuis fed and developed were less attractive to the whitefly Bemisia tabaci, but were more attractive to the parasitoid Encarsia formosa. Female-exposed plants were more repellent to B. tabaci and more attractive to E. formosa than were male-exposed plants. When comparing young- and mature-nymph-exposed plants, the same level of repellence was obtained for B. tabaci, but mature-nymph-exposed plants were more attractive to E. formosa. The repellent effect is attributed to the signaling pathway of abscisic acid, which is upregulated in N. tenuis-exposed plants, whereas the parasitoid attraction was attributed to the activation of the jasmonic acid signaling pathway. Our results demonstrate that all motile stages of N. tenuis can trigger defensive responses in tomato plants, although these responses may be slightly different depending on the stage considered.

Disentangling mite predator-prey relationships by multiplex PCR.

Gut content analysis using molecular techniques can help elucidate predator-prey relationships in situations in which other methodologies are not feasible, such as in the case of trophic interactions between minute species such as mites. We designed species-specific primers for a mite community occurring in Spanish citrus orchards comprising two herbivores, the Tetranychidae Tetranychus urticae and Panonychus citri, and six predatory mites belonging to the Phytoseiidae family; these predatory mites are considered to be these herbivores' main biological control agents. These primers were successfully multiplexed in a single PCR to test the range of predators feeding on each of the two prey species. We estimated prey DNA detectability success over time (DS50), which depended on the predator-prey combination and ranged from 0.2 to 18 h. These values were further used to weight prey detection in field samples to disentangle the predatory role played by the most abundant predators (i.e. Euseius stipulatus and Phytoseiulus persimilis). The corrected predation value for E. stipulatus was significantly higher than for P. persimilis. However, because this 1.5-fold difference was less than that observed regarding their sevenfold difference in abundance, we conclude that P. persimilis is the most effective predator in the system; it preyed on tetranychids almost five times more frequently than E. stipulatus did. The present results demonstrate that molecular tools are appropriate to unravel predator-prey interactions in tiny species such as mites, which include important agricultural pests and their predators.

Tetranychus urticae-triggered responses promote genotype-dependent conspecific repellence or attractiveness in citrus.

The citrus rootstocks sour orange and Cleopatra mandarin display differential resistance against Tetranychus urticae. Sour orange plants support reduced oviposition, growth rates and damage compared with Cleopatra mandarin plants. Jasmonic acid signalling and flavonoid accumulation have been revealed as key mechanisms for the enhanced resistance of sour orange plants. In this study, we observed that the release of T. urticae herbivore-induced plant volatiles (HIPVs) from sour orange plants has a marked repellent effect on conspecific mites associated with the production of the terpenes α-ocimene, α-farnesene, pinene and d-limonene, and the green leaf volatile 4-hydroxy-4-methyl-2-pentanone. By contrast, T. urticae HIPVs from Cleopatra mandarin plants promote conspecific mite attraction associated with an increase in (2-butoxyethoxy) ethanol, benzaldehyde and methyl salicylate levels. HIPVs released from sour orange plants following T. urticae infestation induce resistance in Cleopatra mandarin plants, thereby reducing oviposition rates and stimulating the oxylipin biosynthetic gene lipoxygenase2 (LOX2). Cleopatra HIPVs do not affect the response to T. urticae of these rootstocks. We conclude that sour orange plants promote herbivore-induced resistance in Cleopatra mandarin plants and, despite the weak basal resistance of these rootstocks, herbivore resistance can be induced through the combination of HIPVs, such as α-ocimene and d-limonene.