Herbivore Defense Research Articles

Two leucine-rich repeat receptor-like kinases initiate herbivory defense responses in tea plants

Abstract Leucine-rich repeat receptor-like kinases (LRR-RLKs) have emerged as key regulators of herbivory perception and subsequent defense initiation. While their functions in grass plants have been gradually elucidated, the roles of herbivory-related LRR-RLKs in woody plants remain largely unknown. In this study, we mined the genomic and transcriptomic data of tea plants (Camellia sinensis) and identified a total of 307 CsLRR-RLK members. Phylogenetic analysis grouped these CsLRR-RLKs into 14 subgroups along with their Arabidopsis homologs. Gene structure and conserved domain analyses revealed notable similarities among subgroup members. Among the identified CsLRR-RLKs, we focused on two plasma membrane-localized LRR-RLKs, CsLRR-RLK44 and CsLRR-RLK239, which do not form homodimers or heterodimers with each other. Both respond strongly to herbivory, and their expression patterns significantly correlate with herbivore resistance phenotypes across different tea accessions. CsLRR-RLK44 and CsLRR-RLK239 act upstream of mitogen-activated protein kinase (MPK) cascades and modulate the expression of defense-related MPKs and WRKY transcription factors. Additionally, silencing CsLRR-RLK44 or CsLRR-RLK239 reduced the levels of herbivory-induced jasmonates, thereby weakening the plant resistance to tea geometrid larvae (Ectropis obliqua). Our work is the first to demonstrate that in woody plants, LRR-RLKs are essential for enhancing herbivore resistance through the activation of the canonical signaling, including MPKs, WRKYs and jasmonates. Furthermore, our study extends mechanistic insights into how LRR-RLKs initiate plant defenses from grasses to economically important tree species.

Combined Effects of Heavy Metal and Simulated Herbivory on Leaf Trichome Density in Sunflowers

Trichomes play a key role in both heavy metal tolerance and herbivory defense, and both stressors have been shown to induce increased trichome density. However, the combined effect of these stressors on trichome density in general, and specifically on metal-hyperaccumulating plants, has yet to be examined. The aim of this study was to test the effect of cadmium availability and herbivory on leaf trichome density and herbivore deterrence in the metal hyperaccumulator Helianthus annuus. To test this, H. Annuus plants were grown in control pots or pots inoculated with 10 mg/kg cadmium and were subjected to either no herbivory or simulated herbivory using mechanical damage and foliar jasmonic acid application. Herbivore deterrence was tested in a feeding assay using Spodoptera littoralis caterpillars. Interestingly, while the trichome density of H. annuus increased by 79% or 53.5% under high cadmium availability or simulated herbivory, respectively, it decreased by 26% when the stressors were combined. Furthermore, regardless of cadmium availability, simulated herbivory induced a 40% increase in deterrence of S. littoralis. These findings suggest that the combination of metal availability and herbivory might present excessive stress to hyperaccumulators. Moreover, they suggest that the risk of metal bioaccumulation in phytoremediation can be reduced by simulated herbivory.

Biogenic secondary organic aerosol participates in plant interactions and herbivory defense.

Biogenic secondary organic aerosols (SOAs) can be formed from the oxidation of plant volatiles in the atmosphere. Herbivore-induced plant volatiles (HIPVs) can elicit plant defenses, but whether such ecological functions persist after they form SOAs was previously unknown. Here we show that Scots pine seedlings damaged by large pine weevils feeding on their roots release HIPVs that trigger defenses in neighboring conspecific plants. The biological activity persisted after HIPVs had been oxidized to form SOAs, which was indicated by receivers displaying enhanced photosynthesis, primed volatile defenses, and reduced weevil damage. The elemental composition and quantity of SOAs likely determines their biological functions. This work demonstrates that plant-derived SOAs can mediate interactions between plants, highlighting their ecological significance in ecosystems.

Elemental composition and potential toxicity of the riverine macrophyte Podostemum ceratophyllum Michx. reflects land use in eastern North America

Land use influences surface water quality, often alleviating stoichiometric constraints on primary production and altering biogeochemical cycling. However, land use effects on nutrient content and potential trace metal accumulation in aquatic plants remain unclear, and high concentrations of metals and altered nutrient ratios could impact the health of herbivores and detritivores. We tested for land use effects on nutrient and trace metal accumulation in a widespread riverine macrophyte, Podostemum ceratophyllum, collected from 91 locations from Georgia to Maine, USA in 2014–2016. We quantified carbon (C), nitrogen (N), phosphorus (P), their molar and mass ratios, N and C stable isotopes, and 17 additional elements in dried plants collected from each location to estimate relationships between plant tissue content and watershed land use, which we quantified as agriculture, forest, and development. Decreasing forest cover was correlated with increasing δ15N, Mg, Mn, and P in Podostemum tissue. Increasing urban development was correlated with increasing δ15N, Mg and P, while increasing agriculture was correlated with a decrease in C: P and the concentrations of multiple metals, along with increases in P, Mg and δ15N. Decreases in ratios of N: P and C:P with increasing agriculture and urban development in the watershed indicate more rapid P storage relative to C and N in plant tissue, and increased resource quality of the plant to consumers in these watersheds. We also observed potentially toxic dietary concentrations of some trace metals (B, Cd, Tl, Zn) in plant tissue which could be related to the plant's natural herbivory defense system or to land use. We conclude that land use influences the elemental composition of P. ceratophyllum, and potentially the quality and toxicity of the plant to herbivores and detritivores in eastern North American rivers.

Phenylalanine ammonia-lyases and 4-coumaric acid coenzyme A ligases in Chara braunii, Marchantia polymorpha, and Physcomitrium patens as extant model organisms for plant terrestrialization.

The conquest of land posed severe problems to plants which they had to cope with by adapting biosynthetic capacities. Adaptations to respond to UV irradiation, water loss, pathogen and herbivore defense, and the earth's pull were essential. Chemical compounds alleviating these problems can be synthesized by the phenylpropanoid pathway, the core of which are three enzymes: phenylalanine ammonia-lyase (PAL), cinnamic acid 4-hydroxylase, and 4-coumaric acid coenzyme A-ligase (4CL). The genomes of model organisms, Chara braunii as aquatic alga and the two bryophytes Physcomitrium patens and Marchantia polymorpha, were searched for sequences encoding PAL and 4CL and selected sequences heterologously expressed in Escherichia coli for biochemical characterization. Several possible isoforms were identified for both enzymes in Marchantia polymorpha and Physcomitrium patens, while only one or two isoforms could be retrieved for Chara braunii. Active forms of both enzymes were found in all three organisms, although the catalytic efficiencies varied in a wide range. l-Phenylalanine was accepted as best substrate by all PAL-like enzymes, despite annotations in some cases suggesting different activities. The substrate spectrum of 4CLs was more diverse, but caffeic and/or 4-coumaric acids generally were the best-accepted substrates. Our investigations show that PAL and 4CL, important enzymes for the formation of phenolic compounds, are present and active in extant charophytes and bryophytes as model organisms for the conquest of land.

Patterns of constitutive and induced herbivore defence are complex, but share a common genetic basis in annual and perennial monkeyflower

Abstract Despite multiple ecological and evolutionary hypotheses that predict patterns of phenotypic relationships between plant growth, reproduction and constitutive and/or induced resistance to herbivores, these hypotheses do not make any predictions about the underlying molecular genetic mechanisms that mediate these relationships. We investigated how divergent plant life‐history strategies in the yellow monkeyflower and a life‐history altering locus, DIV1, influence plasticity of phytochemical herbivory resistance traits in response to attack by two herbivore species with different diet breadth. Life‐history strategy (annual vs. perennial) and the DIV1 locus significantly influenced levels of constitutive herbivory resistance, as well as resistance induction following both generalist and specialist herbivory. Perennial plants had higher total levels of univariate constitutive and induced defence than annuals, regardless of herbivore type. Annuals induced less in response to generalist herbivory than did perennials, while induction response was equivalent across the ecotypes for specialist herbivory. The effects of the DIV1 locus on levels of constitutive and induced defence were dependent on genetic background, the annual versus perennial haplotype of DIV1 and herbivore identity. The patterns of univariate induction due to DIV1 were non‐additive and did not always match expectations based on patterns of divergence for annual/perennial parents. For example, perennial plants had higher levels of constitutive and induced defence than did annuals, but when the annual DIV1 was present in the perennial genetic background induction response to herbivory was higher than for the perennial parent lines. Patterns for multivariate defence arsenals generally echoed those of univariate, with annual and perennial monkeyflowers and those with alternative versions of DIV1 differing significantly in constitutive and induced resistance. Like univariate resistance, induced multivariate defence arsenals were affected by herbivore identity. Our results highlight the complexity of the genetic mechanisms underlying plastic response to herbivory. While a genetic locus underlying substantial phenotypic variation in life‐history strategy and constitutive defence also influences defence plasticity, the induction response also depends on genetic background. This result demonstrates the potential for some degree of evolutionary independence between constitutive and induced defence, or induced defence and life‐history strategy, in monkeyflowers. Read the free Plain Language Summary for this article on the Journal blog.

Trichomes and unique gene expression confer insect herbivory resistance in Vitis labrusca grapevines

BackgroundGrapevine (Vitis) is one of the world’s most valuable fruit crops, but insect herbivory can decrease yields. Understanding insect herbivory resistance is critical to mitigating these losses. Vitis labrusca, a wild North American grapevine species, has been leveraged in breeding programs to generate hybrid grapevines with enhanced abiotic and biotic stress resistance, rendering it a valuable genetic resource for sustainable viticulture. This study assessed the resistance of V. labrusca acc. ‘GREM4’ and Vitis vinifera cv. ‘PN40024’ grapevines to Popillia japonica (Japanese beetle) herbivory and identified morphological and genetic adaptations underlying this putative resistance.Results‘GREM4’ displayed greater resistance to beetle herbivory compared to ‘PN40024’ in both choice and no-choice herbivory assays spanning periods of 30 min to 19 h. ‘GREM4’ had significantly higher average leaf trichome densities than ‘PN40024’ and beetles preferred to feed on the side of leaves with fewer trichomes. When leaves from each species that specifically did not differ in trichome densities were fed on by beetles, significantly less leaf area was damaged in ‘GREM4’ (3.29mm2) compared to ‘PN40024’ (9.80mm2), suggesting additional factors beyond trichomes contributed to insect herbivory resistance in ‘GREM4’. Comparative transcriptomic analyses revealed ‘GREM4’ exhibited greater constitutive (0 h) expression of defense response and secondary metabolite biosynthesis genes compared to ‘PN40024’, indicative of heightened constitutive defenses. Upon herbivory, ‘GREM4’ displayed a greater number of differentially expressed genes (690) compared to ‘PN40024’ (502), suggesting a broader response. Genes up-regulated in ‘GREM4’ were enriched in terpene biosynthesis, flavonoid biosynthesis, phytohormone signaling, and disease defense-related functions, likely contributing to heighted insect herbivory defense, while genes differentially expressed in ‘PN40024’ under herbivory were enriched in xyloglucan, cell wall formation, and calcium ion binding. The majority of genes implicated in insect herbivory defense were orthologs with specific expression patterns in ‘GREM4’ and ‘PN40024’, but some paralogous and genome-specific genes also likely contributed to conferring resistance.ConclusionsOur findings suggest that ‘GREM4’ insect herbivory resistance was attributed to a combination of factors, including trichomes and unique constitutive and inducible expression of genes implicated in terpene, flavonoid, and phenylpropanoid biosynthesis, as well as pathogen defense.

Ethylene: A Modulator of the Phytohormone-Mediated Insect Herbivory Network in Plants.

Plants have evolved to establish insect herbivory defences by modulating their metabolism, growth, and development. Precise networks of phytohormones are essential to induce those herbivory defences. Gaseous phytohormone ET plays an important role in forming herbivory defences. Its role in insect herbivory is not fully understood, but previous studies have shown that it can both positively and negatively regulate herbivory. This review presents recent findings on crosstalk between ET and other phytohormones in herbivory responses. Additionally, the use of exogenous ETH treatment to induce ET in response to herbivory is discussed.

Comparative venation costs of monocotyledon and dicotyledon species in the eastern Colorado steppe.

Leaf vein network cost (total vein surface area per leaf volume) for major veins and vascular bundles did not differ between monocot and dicot species in 21 species from the eastern Colorado steppe. Dicots possessed significantly larger minor vein networks than monocots. Across the tree of life, there is evidence that dendritic vascular transport networks are optimized, balancing maximum speed and integrity of resource delivery with minimal resource investment in transport and infrastructure. Monocot venation, however, is not dendritic, and remains parallel down to the smallest vein orders with no space-filling capillary networks. Given this departure from the "optimized" dendritic network, one would assume that monocots are operating at a significant energetic disadvantage. In this study, we investigate whether monocot venation networks bear significantly greater carbon/construction costs per leaf volume than co-occurring dicots in the same ecosystem, and if so, what physiological or ecological advantage the monocot life form possesses to compensate for this deficit. Given that venation networks could also be optimized for leaf mechanical support or provide herbivory defense, we measured the vascular system of both monocot and dicots at three scales to distinguish between leaf investment in mechanical support (macroscopic vein), total transport and capacitance (vascular bundle), or exclusively water transport (xylem) for both parallel and dendritic venation networks. We observed that vein network cost (total vein surface area per leaf volume) for major veins and vascular bundles was not significantly different between monocot species and dicot species. Dicots, however, possess significantly larger minor vein networks than monocots. The 19 species subjected to gas-exchange measurement in the field displayed a broad range of Amax and but demonstrated no significant relationships with any metric of vascular network size in major or minor vein classes. Given that monocots do not seem to display any leaf hydraulic disadvantage relative to dicots, it remains an important research question why parallel venation (truly parallel, down to the smallest vessels) has not arisen more than once in the history of plant evolution.

Fungal endophytes can modulate plant invasion.

Symbiotic organisms may contribute to a host plant's success or failure to grow, its ability to maintain viable populations, and potentially, its probability of establishment and spread outside its native range. Intercellular and intracellular microbial symbionts that are asymptomatic in their plant host during some or all of their life cycle - endophytes - can form mutualistic, commensal, or pathogenic relationships, and sometimes novel associations with alien plants. Fungal endophytes are likely the most common endosymbiont infecting plants, with life-history, morphological, physiological, and plant-symbiotic traits that are distinct from other endophytic guilds. Here, we review the community dynamics of fungal endophytes during the process of plant invasion, and how their functional role may shift during the different stages of invasion: transport, introduction (colonisation), establishment, and spread. Each invasion stage presents distinct ecological filters that an alien plant must overcome to advance to the subsequent stage of invasion. Endophytes can alternately aid the host in overcoming stage-specific filters, or contribute to the barriers imposed by filters (e.g. biotic resistance), thereby affecting invasion pathways. A few fungi can be transported as seed endophytes from their native range and be vertically transmitted to future generations in the non-native range, especially in graminoids. In other plant groups, alien plants mostly acquire endophytes via horizontal transmission from the invaded plant community, and the host endophyte community is shaped by host filtering and biogeographic factors (e.g. dispersal limitation, environmental filtering). Endophytes infecting alien plants (both those transported with their host and those accumulated in the non-native range) may influence invasion success by affecting plant growth, reproduction, environmental tolerance, and pathogen and herbivory defences; however, the direction and magnitude of these effects can be contingent upon the host identity, life stage, ecological conditions, and invasion stage. This context dependence may cause endophytic fungi to shift to a non-endophytic (e.g. pathogenic) functional life stage in the same or different hosts, which can modify alien-native plant community dynamics. We conclude by identifying paths in which alien hosts can exploit the context dependency of endophyte function in novel abiotic and biotic conditions and at the different stages of invasion.

Duplicated Copy Number Variant of the Maize 9-Lipoxygenase ZmLOX5 Improves 9,10-KODA-Mediated Resistance to Fall Armyworms.

Extensive genome structure variations, such as copy number variations (CNVs) and presence/absence variations, are the basis for the remarkable genetic diversity of maize; however, the effect of CNVs on maize herbivory defense remains largely underexplored. Here, we report that the naturally occurring duplication of the maize 9-lipoxygenase gene ZmLOX5 leads to increased resistance of maize to herbivory by fall armyworms (FAWs). Previously, we showed that ZmLOX5-derived oxylipins are required for defense against chewing insect herbivores and identified several inbred lines, including Yu796, that contained duplicated CNVs of ZmLOX5, referred to as Yu796-2×LOX5. To test whether introgression of the Yu796-2×LOX5 locus into a herbivore-susceptible B73 background that contains a single ZmLOX5 gene is a feasible approach to increase resistance, we generated a series of near-isogenic lines that contained either two, one, or zero copies of the Yu796-2×LOX5 locus in the B73 background via six backcrosses (BC6). Droplet digital PCR (ddPCR) confirmed the successful introgression of the Yu796-2×LOX5 locus in B73. The resulting B73-2×LOX5 inbred line displayed increased resistance against FAW, associated with increased expression of ZmLOX5, increased wound-induced production of its primary oxylipin product, the α-ketol, 9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid (9,10-KODA), and the downstream defense hormones regulated by this molecule, 12-oxo-phytodienoic acid (12-OPDA) and abscisic acid (ABA). Surprisingly, wound-induced JA-Ile production was not increased in B73-2×LOX5, resulting from the increased JA catabolism. Furthermore, B73-2×LOX5 displayed reduced water loss in response to drought stress, likely due to increased ABA and 12-OPDA content. Taken together, this study revealed that the duplicated CNV of ZmLOX5 quantitively contributes to maize antiherbivore defense and presents proof-of-concept evidence that the introgression of naturally occurring duplicated CNVs of a defensive gene into productive but susceptible crop varieties is a feasible breeding approach for enhancing plant resistance to herbivory and tolerance to abiotic stress.

Blocking then stinging as a case of two-step evolution of defensive cage architectures in herbivore-driven ecosystems.

Dense branching and spines are common features of plant species in ecosystems with high mammalian herbivory pressure. While dense branching and spines can inhibit herbivory independently, when combined, they form a powerful defensive cage architecture. However, how cage architecture evolved under mammalian pressure has remained unexplored. Here we show how dense branching and spines emerged during the age of mammalian radiation in the Combretaceae family and diversified in herbivore-driven ecosystems in the tropics. Phylogenetic comparative methods revealed that modern plant architectural strategies defending against large mammals evolved via a stepwise process. First, dense branching emerged under intermediate herbivory pressure, followed by the acquisition of spines that supported higher speciation rates under high herbivory pressure. Our study highlights the adaptive value of dense branching as part of a herbivore defence strategy and identifies large mammal herbivory as a major selective force shaping the whole plant architecture of woody plants.

Effect of residence time on trait evolution in invasive plants: review and meta-analysis

The success of invasive species is often attributed to rapid post-introduction evolution, due to novel selection pressures at the introduced range. However, evolutionary shifts in invasion-promoting traits can also take place within the introduced range over time. Here, we first present a review of the proposed hypotheses regarding the selection pressures and trait divergence along gradients of invasion history and the studies that examined them. In addition, we present the results of a meta-analysis aimed to provide a more general overview of current knowledge on trait evolution with time since introduction. Invasion-promoting traits, including growth, competitive ability and dispersal ability, were proposed to decline in more established populations with a long invasion history due to the attenuation of selection pressures, such as enemy release or interspecific competition, while herbivore defence was suggested to increase. Our meta-analysis results reveal a general indication for the evolution of invasive plants with residence time for most of the studied traits. However, this divergence did not have a consistent direction in most traits, except for growth, which, in contrast with our prediction, increased with residence time. The lack of empirical support for the predicted change in most of the studied traits over time suggests trait evolution might be affected by other context-dependent factors such as climatic gradients along invasion routes. Similarly, the increased allocation to size in older and more established populations may be driven by increased conspecific competition pressure experienced in these populations. The general temporal effect found in our meta-analysis stresses the need to consider population age when comparing attributes of invasive plants between native and invasive ranges. Moreover, the increased size of invasive plants in older populations, suggests that the dominance of these plants might not attenuate with time since introduction, thus highlighting the need to further explore the long-term dynamics between invasive plants and their recipient native communities.

ApCPS2 contributes to medicinal diterpenoid biosynthesis and defense against insect herbivore in Andrographis paniculata

Kalmegh (Andrographis paniculata) spatiotemporally produces medicinally-important ent-labdane-related diterpenoids (ent-LRDs); andrographolide (AD), 14-deoxy-11,12-didehydroandrographolide (DDAD), neoandrographolide (NAD). ApCPS1 and ApCPS2, the ent-copalyl pyrophosphate (ent-CPP)-producing class II diterpene synthases (diTPSs) were identified, but their contributions to ent-CPP precursor supply for ent-LRD biosynthesis were not well understood. Here, we characterized ApCPS4, an additional ent-CPP-forming diTPS. Further, we elucidated in planta function of the ent-CPP-producing diTPSs (ApCPS1,2,4) by integrating transcript-metabolite co-profiles, biochemical analysis and gene functional characterization. ApCPS1,2,4 localized to the plastids, where diterpenoid biosynthesis occurs in plants, but ApCPS1,2,4 transcript expression patterns and ent-LRD contents revealed a strong correlation of ApCPS2 expression and ent-LRD accumulation in kalmegh. ApCPS1,2,4 upstream sequences differentially activated β-glucuronidase (GUS) in Arabidopsis and transiently-transformed kalmegh. Similar to higher expression of ApCPS1 in kalmegh stem, ApCPS1 upstream sequence activated GUS in stem/hypocotyl of Arabidopsis and kalmegh. However, ApCPS2,4 upstream sequences weakly activated GUS expression in Arabidopsis, which was not well correlated with ApCPS2,4 transcript expression in kalmegh tissues. Whereas, ApCPS2,4 upstream sequences could activate GUS expression at a considerable level in kalmegh leaf and roots/calyx, respectively, suggesting the involvement of transcriptional regulator(s) of ApCPS2,4 that might participate in kalmegh-specific diterpenoid pathway. Interestingly, ApCPS2-silenced kalmegh showed a drastic reduction in AD, DDAD and NAD contents and compromised defense against insect herbivore Spodoptera litura. However, ent-LRD contents and herbivore defense in ApCPS1 or ApCPS4-silenced plants remained largely unaltered. Overall, these results suggested an important role of ApCPS2 in producing ent-CPP for medicinal ent-LRD biosynthesis and defense against insect herbivore.

Plant physical defenses contribute to a latitudinal gradient in resistance to insect herbivory within a widespread perennial grass.

Herbivore pressure can vary across the range of a species, resulting in different defensive strategies. If herbivory is greater at lower latitudes, plants may be better defended there, potentially driving a latitudinal gradient in defense. However, relationships that manifest across the entire range of a species may be confounded by differences within genetic subpopulations, which may obscure the drivers of these latitudinal gradients. We grew plants of the widespread perennial grass Panicum virgatum in a common garden that included genotypes from three genetic subpopulations spanning an 18.5° latitudinal gradient. We then assessed defensive strategies of these plants by measuring two physical resistance traits-leaf mass per area (LMA) and leaf ash, a proxy for silica-and multiple measures of herbivory by caterpillars of the generalist herbivore fall armyworm (Spodoptera frugiperda). Across all genetic subpopulations, low-latitude plants experienced less herbivory than high-latitude plants. Within genetic subpopulations, however, this relationship was inconsistent-the most widely distributed and phenotypically variable subpopulation (Atlantic) exhibited more consistent latitudinal trends than either of the other two subpopulations. The two physical resistance traits, LMA and leaf ash, were both highly heritable and positively associated with resistance to different measures of herbivory across all subpopulations, indicating their importance in defense against herbivores. Again, however, these relationships were inconsistent within subpopulations. Defensive gradients that occur across the entire species range may not arise within localized subpopulations. Thus, identifying the drivers of latitudinal gradients in herbivory defense may depend on adequately sampling the diversity within a species.

Silicon-mediated herbivore defence in a pasture grass under reduced and Anthropocene levels of CO2.

The uptake and accumulation of silicon (Si) in grass plants play a crucial role in alleviating both biotic and abiotic stresses. Si supplementation has been reported to increase activity of defence-related antioxidant enzyme, which helps to reduce oxidative stress caused by reactive oxygen species (ROS) following herbivore attack. Atmospheric CO2 levels are known to affect Si accumulation in grasses; reduced CO2 concentrations increase Si accumulation whereas elevated CO2 concentrations often decrease Si accumulation. This can potentially affect antioxidant enzyme activity and subsequently insect herbivory, but this remains untested. We examined the effects of Si supplementation and herbivory by Helicoverpa armigera on antioxidant enzyme (catalase, CAT; superoxide dismutase, SOD; and ascorbate peroxidase, APX) activity in tall fescue grass (Festuca arundinacea) grown under CO2 concentrations of 200, 410, and 640 ppm representing reduced, ambient, and elevated CO2 levels, respectively. We also quantified foliar Si, carbon (C), and nitrogen (N) concentrations and determined how changes in enzymes and elemental chemistry affected H. armigera relative growth rates and plant consumption. Rising CO2 concentrations increased plant mass and foliar C but decreased foliar N and Si. Si supplementation enhanced APX and SOD activity under the ranging CO2 regimes. Si accumulation and antioxidant enzyme activity were at their highest level under reduced CO2 conditions and their lowest level under future levels of CO2. The latter corresponded with increased herbivore growth rates and plant consumption, suggesting that some grasses could become more susceptible to herbivory under projected CO2 conditions.

Epidermal bladder cells as a herbivore defense mechanism

The aerial surfaces of quinoa (Chenopodium quinoa) and common ice plant (Mesembryanthemum crystallinum) are covered with a layer of epidermal bladder cells (EBCs), which are modified non-glandular trichomes previously considered to be key to the extreme salt and drought tolerance of these plants. Here, however, we find that EBCs of these plants play only minor roles, if any, in abiotic stress tolerance and in fact are detrimental under conditions of water deficit. We report that EBCs instead function as deterrents to a broad range of generalist arthropod herbivores, through their combined function of forming both a chemical and a physical barrier, and they also serve a protective function against a phytopathogen. Our study overturns current models that link EBCs to salt and drought tolerance and assigns new functions to these structures that might provide novel possibilities for protecting crops from arthropod pests.

Assessing the adaptive role of cannabidiol (CBD) in Cannabis sativa defense against cannabis aphids.

Cannabis sativa is known for having unique specialized or secondary metabolites, cannabinoids that are derived from an extension of the terpene pathway in the Cannabis lineage and includes more than 100 other similar metabolites. Despite the assumption that cannabinoids evolved as novel herbivory defense adaptations, there is limited research addressing the role of cannabinoids in C. sativa responses to insect herbivores. Here we investigated the role of cannabidiol (CBD), the predominant cannabinoid in hemp, in plant defense against cannabis aphid (Phorodon cannabis), one of the most damaging pests of hemp. We hypothesize that insect feeding may induce changes in cannabinoids as an adaptive strategy for defense. We found that mean fecundity, net reproductive rate (R0) and adult longevity of cannabis aphids was reduced on the high cannabinoid cultivar compared to the low- cannabinoid cultivar in whole plant assays. In contrast, supplementation of CBD in artificial feeding assays increased aphid fecundity from day 1 to day 3. Additionally, aphid feeding did not impact cannabinoid levels in leaf tissues with the exception of Δ9-tetrahydrocannabinol (THC). This suggests that other cannabinoids and/or metabolites such as terpenes are causing the observed decrease in aphid performance in the whole plant assays. In addition to cannabinoids, C. sativa also possesses a range of defense mechanisms via phytohormone signaling pathways that are well described in other plant species. Indeed, cannabis aphid feeding significantly increased levels of the major phytohormones, salicylic acid, jasmonic acid, and abscisic acid, which are known to be involved in plant defense responses against aphid species. These results highlight the interplay between cannabinoid synthesis and phytohormone pathways and necessitate further investigation into this complex interaction.

Silicon-based anti-herbivore defense in tropical tree seedlings.

Silicon-based defenses deter insect herbivores in many cultivated and wild grass species. Furthermore, in some of these species, silicon (Si) uptake and defense can be induced by herbivory. Tropical trees also take up Si and leaf Si concentrations vary greatly across and within species. As herbivory is a major driver of seedling mortality and niche differentiation of tropical tree species, understanding anti-herbivore defenses is pivotal. Yet, whether silicon is a constitutive and inducible herbivory defense in tropical forest tree species remains unknown. We grew seedlings of eight tropical tree species in a full factorial experiment, including two levels of plant-available soil Si concentrations (-Si/+Si) and a simulated herbivory treatment (-H/+H). The simulated herbivory treatment was a combination of clipping and application of methyl jasmonate. We then carried out multiple-choice feeding trials, separately for each tree species, in which leaves of each treatment combination were offered to a generalist caterpillar (Spodoptera frugiperda). Leaf damage was assessed. Three species showed a significant decrease in leaf damage under high compared to low Si conditions (by up to 72%), consistent with our expectation of Si-based defenses acting in tropical tree species. In one species, leaf damage was increased by increasing soil Si and in four species, no effect of soil Si on leaf damage was observed. Opposite to our expectation of Si uptake and defense being inducible by herbivory damage, simulated herbivory increased leaf damage in two species. Furthermore, simulated herbivory reduced Si concentrations in one species. Our results showed that tropical tree seedlings can be better defended when growing in Si-rich compared to Si-poor soils, and that the effects of Si on plant defense vary strongly across species. Furthermore, Si-based defenses may not be inducible in tropical tree species. Overall, constitutive Si-based defense should be considered part of the vast array of anti-herbivore defenses of tropical tree species. Our finding that Si-based defenses are highly species-specific combined with the fact that herbivory is a major driver of mortality in tropical tree seedling, suggests that variation in soil Si concentrations may have pervasive consequences for regeneration and performance across tropical tree species.

Knocking down a DNA demethylase gene affects potato plant defense against a specialist insect herbivore.

DNA demethylase (DML) is involved in plant development and responses to biotic and abiotic stresses; however, its role in plant-herbivore interaction remains elusive. Here, we found that herbivory by the potato tuber moth, Phthorimaea operculella, rapidly induced the genome-wide DNA methylation and accumulation of DML gene transcripts in potato plants. Herbivory induction of DML transcripts was suppressed in jasmonate-deficient plants, whereas exogenous application of methyl jasmonate (MeJA) improved DML transcripts, indicating that the induction of DML transcripts by herbivory is associated with jasmonate signaling. Moreover, P. operculella larvae grew heavier on DML gene (StDML2) knockdown plants than on wild-type plants, and the decreased biosynthesis of jasmonates in the former may be responsible for this difference, since the larvae feeding on these two genotypes supplemented with MeJA showed similar growth. In addition, P. operculella adult moths preferred to oviposit on StDML2 knockdown plants than on wild-type plants, which was associated with the reduced emission of β-caryophyllene in the former. In addition, supplementing β-caryophyllene to these two genotypes further disrupted moths' oviposit choice preference for them. Interestingly, in StDML2 knockdown plants, hypermethylation was found at the promoter regions for the key genes StAOS and StAOC in the jasmonate biosynthetic pathway, as well as for the key gene StTPS12 in β-caryophyllene production. Our findings suggest that knocking down StDML2 can affect herbivore defense via jasmonate signaling and defense compound production in potato plants.