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Species-specific effects of oviposition and larval herbivory on N. attenuata’s transcriptional regulation. (a) Principle component analysis (PCA) of transcript profiles of N. attenuata in response to either S. exigua (Se) or M. sexta (Ms) oviposition and/or larval feeding. The legend above presents all symbols used to depict all treatments in the 3 × 3 factorial design and treatment replicates are outlined by 68% confidence intervals. (b) Venn diagram comparing the numbers of genes that were regulated in the microarray analysis by either S. exigua or M. sexta larval feeding (relative to control plants) and those significantly different between the two (at P < 0.05 after correction for false discovery rate and a log2-fold change (FC) > 1). (c) Scatter plots of the log2-FC in gene expression in response to larval feeding by M. sexta and S. exigua. (d) Scatter plots of the log2-FC in gene expression in response to larval feeding on plants oviposited and non-oviposited by conspecific moths (left: M. sexta, right: S. exigua). (e) Hierarchical clustering of the log2FC in gene expression (relative to control plants) of all feeding-induced treatments.
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Oviposition by lepidopteran herbivores on Nicotiana attenuata primes plant defence responses that are induced by the feeding larvae. While oviposition by both the generalist Spodoptera exigua and the specialist Manduca sexta primes the production of defensive phenylpropanoids, their larvae are differentially affected. We investigate here the impact...
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Nitrogen (N) and carbon(C) metabolisms in plants were investigated to assess different responses of Bt and non-Bt rice to different N treatments. T2A-1 (Bt rice variety) inserted with Cry2A* protein to resist Lepidoptera and its parental line MH63 was adopted in this study. The total N accumulation presented no statistical difference. But nitrogen...
Citations
... Samples for the phytohormone analyses were collected from trees that were also used for the qPCR analysis, i.e., from an experiment independent of that used for the RNA sequencing analysis. Phytohormone extraction and analyses were conducted following the methods described by Bandoly et al. (2016) and Drok et al. (2018). In short, ethyl acetate (spiked with deuterated phytohormones as internal standards) was used as extraction buffer. ...
... Other angiosperm plants such as A. thaliana (Little et al. 2007, Valsamakis et al. 2022, bittersweet nightshade Solanum dulcamara L. (Geuss et al. 2017) and tobacco plants (Nicotiana attenuata Torr. ex S. Watson) (Drok et al. 2018) have shown a considerable number of DEGs 1-3 days after egg deposition. Similar to the response of pine to sawfly eggs, egg-induced differential expression of genes in elm had almost reverted to the control level by the end of the egg phase (Altmann et al. 2018). ...
... However, an overlap of egg-and feedingresponsive genes was also found in A. thaliana (Valsamakis et al. 2022), in black mustard plants (Brassica nigra L. W. D. J. Koch) (Bonnet et al. 2017) and in tobacco plants (N. attenuata) (Drok et al. 2018) infested with eggs or larvae of lepidopteran species, which do not damage the leaf tissue during oviposition. Furthermore, a Generally Applicable Gene set Enrichment analysis of four angiosperm species treated with insect eggs and larval feeding also revealed a large overlap of insect egg-and feeding-induced responses . ...
Plants can improve their resistance against feeding damage by insects if they have perceived insect egg deposition prior to larval feeding. Molecular analyses of these egg-mediated defence mechanisms have until now focused on angiosperm species. It is unknown how the transcriptome of a gymnosperm species responds to insect eggs and subsequent larval feeding. Scots pine (Pinus sylvestris) is known to improve its defences against larvae of the herbivorous sawfly Diprion pini if it has previously received sawfly eggs. Here, we analysed the transcriptomic and phytohormonal responses of Scots pine needles to D. pini eggs (E-pine), larval feeding (F-pine), and to both eggs and larval feeding (EF-pine). Pine showed strong transcriptomic responses to sawfly eggs and-as expected-to larval feeding. Many egg-responsive genes were also differentially expressed in response to feeding damage, and these genes play an important role in biological processes related to cell wall modification, cell death, and jasmonic acid signalling. EF-pine showed fewer transcriptomic changes than F-pine, whereas EF-treated angiosperm species studied so far showed more transcriptional changes to the initial phase of larval feeding than only feeding-damaged F-angiosperms. However, as with responses of EF-angiosperms, EF-pine showed higher salicylic acid concentrations than F-pine. Based on the considerable overlap of the transcriptomes of E- and F-pine, we suggest that the weaker transcriptomic response of EF-pine than F-pine to larval feeding damage is compensated by the strong, egg-induced response, which might result in maintained pine defences against larval feeding.
... The up-regulation of defense-related pathways, including plant hormone signal transduction, was observed in switchgrass in response to fall armyworm [21]. JA and ET accumulation were observed in plants after infestation by a pest insect [57][58][59][60]. In this study, one lipoxygenase gene involved in linoleic acid metabolism was induced by pest attack. ...
Sugarcane is the most important sugar crop, contributing ≥80% to total sugar production around the world. Spodoptera frugiperda is one of the main pests of sugarcane, potentially causing severe yield and sugar loss. The identification of key defense factors against S. frugiperda herbivory can provide targets for improving sugarcane resistance to insect pests by molecular breeding. In this work, we used one of the main sugarcane pests, S. frugiperda, as the tested insect to attack sugarcane. Integrated transcriptome and metabolomic analyses were performed to explore the changes in gene expression and metabolic processes that occurred in sugarcane leaf after continuous herbivory by S. frugiperda larvae for 72 h. The transcriptome analysis demonstrated that sugarcane pest herbivory enhanced several herbivory-induced responses, including carbohydrate metabolism, secondary metabolites and amino acid metabolism, plant hormone signaling transduction, pathogen responses, and transcription factors. Further metabolome analysis verified the inducement of specific metabolites of amino acids and secondary metabolites by insect herbivory. Finally, association analysis of the transcriptome and metabolome by the Pearson correlation coefficient method brought into focus the target defense genes against insect herbivory in sugarcane. These genes include amidase and lipoxygenase in amino acid metabolism, peroxidase in phenylpropanoid biosynthesis, and pathogenesis-related protein 1 in plant hormone signal transduction. A putative regulatory model was proposed to illustrate the sugarcane defense mechanism against insect attack. This work will accelerate the dissection of the mechanism underlying insect herbivory in sugarcane and provide targets for improving sugarcane variety resistance to insect herbivory by molecular breeding.
... 21,22 Volatiles of herbivore-attacked plants can also prime herbivory-induced defense in adjacent plants. 23 Evidence of the inherited responses to herbivory comes from research with wild radish (Raphanus raphanistrun), where insect-damaged plants produced more resistant seedlings than undamaged plants. 24 In the case of Arabidopsis, priorgeneration feeding by Pieris rapae (white cabbage butterfly) reduced caterpillar weight gain more than MeJA treatment and mechanical damage, in addition to P. rapae, only S. exigua showed reduced performance on plants that were exposed to P. rapae herbivory in the previous generation, these results confirmed the fact that there is species-specific variation in lepidopteran sensitivity to Arabidopsis defenses. ...
Herbivore-induced defense responses are often specific, whereas plants could induce distinct defense responses corresponding to infestation by different herbivorous insects. Brown plant hopper (BPH) Nilaparvata lugens, a phloem-feeding insect, and rice leaf folder (LF) Cnaphalocrocis medinalis, a chewing insect, are both specialist herbivores on rice. To characterize the distinct resistance primed by prior damage to these two specialist herbivores, we challenged rice plants with two herbivores during vegetative growth of parent plants and assessed plant resistance in subsequent ratoons. Here, we show that LF and BPH induce different suites of defense responses in parent rice plants, LF induced higher level of JA accumulation and OsAOS, OsCOI1 transcripts, while BPH induced higher accumulation of SA and OsPAL1 transcripts. Moreover, an apparent loss of LF resistance was observed in OsAOS, OsCOI1 RNAi lines. Ratoon plants generated from parents receiving prior LF infestation exhibited higher jasmonic acid (JA) levels and elevated levels of transcripts of defense-related genes associated with JA signaling, while ratoon generated from parents receiving prior BPH infestation exhibited higher salicylic acid (SA) levels and elevated levels of transcripts of defense-related genes associated with SA signaling. Moreover, previous LF infestation obviously elevated ratoons resistance to LF, while previous infestation by BPH led to enhanced resistance in ratoons to BPH. Pre-priming of ratoons defense to LF was significantly reduced in OsAOS and OsCOI1 RNAi plant, but silencing OsAOS and OsCOI1 did not attenuate ratoons resistance to BPH. These results suggest that infestation of two specialist herbivores with different feeding styles in parent crop led to distinct defense responses in subsequent rations, and the acquired resistance to LF in ratoons is associated with priming of jasmonic acid-dependent defense responses.
... These responses were shown to be dependent on ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), ISOCHORISMATE SYN-THASE 1/SALICYLIC ACID INDUCTION DEFICIENT 2 (ICS1/SID2) and, partially, NONEXPRESSER OF PR GENES 1 (NPR1), which are known signaling components of plant defense responses against biotrophic pathogens [26]. Transcriptomic studies in different plants species have confirmed that insect oviposition induces genes associated with SA-and ROS-mediated immune responses and PR1 gene expression [15,24,[27][28][29][30][31][32][33], including in B. nigra and B. rapa [21,34,35]. Further, it has been suggested that there is a conserved transcriptional response amongst different plant-insect egg interactions [36]. ...
Background
Cabbage white butterflies (Pieris spp.) can be severe pests of Brassica crops such as Chinese cabbage, Pak choi (Brassica rapa) or cabbages (B. oleracea). Eggs of Pieris spp. can induce a hypersensitive response-like (HR-like) cell death which reduces egg survival in the wild black mustard (B. nigra). Unravelling the genetic basis of this egg-killing trait in Brassica crops could improve crop resistance to herbivory, reducing major crop losses and pesticides use. Here we investigated the genetic architecture of a HR-like cell death induced by P. brassicae eggs in B. rapa.
Results
A germplasm screening of 56 B. rapa accessions, representing the genetic and geographical diversity of a B. rapa core collection, showed phenotypic variation for cell death. An image-based phenotyping protocol was developed to accurately measure size of HR-like cell death and was then used to identify two accessions that consistently showed weak (R-o-18) or strong cell death response (L58). Screening of 160 RILs derived from these two accessions resulted in three novel QTLs for Pieris brassicae-induced cell death on chromosomes A02 (Pbc1), A03 (Pbc2), and A06 (Pbc3). The three QTLs Pbc1–3 contain cell surface receptors, intracellular receptors and other genes involved in plant immunity processes, such as ROS accumulation and cell death formation. Synteny analysis with A. thaliana suggested that Pbc1 and Pbc2 are novel QTLs associated with this trait, while Pbc3 also contains an ortholog of LecRK-I.1, a gene of A. thaliana previously associated with cell death induced by a P. brassicae egg extract.
Conclusions
This study provides the first genomic regions associated with the Pieris egg-induced HR-like cell death in a Brassica crop species. It is a step closer towards unravelling the genetic basis of an egg-killing crop resistance trait, paving the way for breeders to further fine-map and validate candidate genes.
... Similarly, significantly enhanced SA levels were detected in A. thaliana when laden with eggs by the butterfly Pieris brassicae and subsequently damaged by its hatching larvae for 24 h (Lortzing et al. 2019;Valsamakis et al. 2020). In contrast, moth egg depositions upon the annual species Nicotiana attenuata and the perennial S. dulcamara did not affect SA levels in subsequently larval feeding-damaged leaves (Drok et al. 2018). Thus, the SA response of plants to egg deposition and subsequent larval feeding damage varies according to the interacting plant and insect species. ...
Main conclusion
Elms, which have received insect eggs as a ‘warning’ of larval herbivory, enhance their anti-herbivore defences by accumulating salicylic acid and amplifying phenylpropanoid-related transcriptional and metabolic responses to hatching larvae.
Abstract
Plant responses to insect eggs can result in intensified defences against hatching larvae. In annual plants, this egg-mediated effect is known to be associated with changes in leaf phenylpropanoid levels. However, little is known about how trees—long-living, perennial plants—improve their egg-mediated, anti-herbivore defences. The role of phytohormones and the phenylpropanoid pathway in egg-primed anti-herbivore defences of a tree species has until now been left unexplored. Using targeted and untargeted metabolome analyses we studied how the phenylpropanoid pathway of Ulmus minor responds to egg-laying by the elm leaf beetle and subsequent larval feeding. We found that when compared to untreated leaves, kaempferol and quercetin concentrations increased in feeding-damaged leaves with prior egg deposition, but not in feeding-damaged leaves without eggs. PCR analyses revealed that prior insect egg deposition intensified feeding-induced expression of phenylalanine ammonia lyase ( PAL ), encoding the gateway enzyme of the phenylpropanoid pathway . Salicylic acid (SA) concentrations were higher in egg-treated, feeding-damaged leaves than in egg-free, feeding-damaged leaves, but SA levels did not increase in response to egg deposition alone—in contrast to observations made of Arabidopsis thaliana . Our results indicate that prior egg deposition induces a SA-mediated response in elms to feeding damage. Furthermore, egg deposition boosts phenylpropanoid biosynthesis in subsequently feeding-damaged leaves by enhanced PAL expression, which results in the accumulation of phenylpropanoid derivatives. As such, the elm tree shows similar, yet distinct, responses to insect eggs and larval feeding as the annual model plant A. thaliana.
... These responses were shown to be dependent on EDS1, ISOCHORISMATE SYNTHASE 1/SALICYLIC ACID INDUCTION DEFICIENT 2 (ICS1/SID2) and, partially, (NONEXPRESSER OF PR GENES 1) (NPR1), which are known signaling components of plant defense responses against biotrophic pathogens [26]. Transcriptomics studies in different plants species have con rmed that insect oviposition induces genes associated with SA-and ROS-mediated immune responses and PR1 gene expression [15,24,[27][28][29][30][31][32][33], including in B. nigra and B. rapa [21,34,35]. It is suggested that there is a conserved transcriptional response amongst different plant-insect eggs interactions [36]. ...
Background
Cabbage white butterflies (Pieris spp.) can be severe pests of Brassica crops such as Chinese cabbage, Pak choi (Brassica rapa) or cabbages (B. oleracea). Eggs of Pieris spp. can induce a hypersensitive response-like (HR-like) cell death which reduces egg survival in the wild black mustard (B. nigra). Unravelling the genetic basis of this egg-killing trait in Brassica crops could improve crop resistance to herbivory, reducing major crop losses and pesticides use. Here we investigated the genetic architecture of a HR-like cell death induced by P. brassicae eggs in B. rapa.
Results
A germplasm screening of B. rapa 56 accessions, representing the genetic and geographical diversity of a B. rapa core collection, showed phenotypic variation for cell death. An image-based phenotyping protocol was developed to accurately measure size of HR-like cell death and was then used to identify two accessions that consistently showed weak (R-o-18) or strong cell death response (L58). Screening of 160 RILs derived from these two accessions resulted in three novel QTLs for Pieris brassicae-induced cell death on chromosomes A02 (Pbc1), A03 (Pbc2), and A06 (Pbc3). The three QTLs Pbc1-3 contain cell surface receptors, intracellular receptors and other genes involved in plant immunity processes, such as ROS accumulation and cell death formation. Synteny analysis with A. thaliana suggested that Pbc1 and Pbc2 are novel QTLs associated with this trait, while Pbc3 contains also LecRK-I.1, a gene of A. thaliana previously associated with cell death induced by a P. brassicae egg extract.
Conclusions
This study provides the first genomic regions associated with the Pieris egg-induced HR-like cell death in a Brassica crop species. It is a step closer towards unravelling the genetic basis of an egg-killing crop resistance trait, paving the way for breeders to further fine-map and validate candidate genes.
... These interactions have been responsible for the development of an extensive machinery of metabolization and detoxification that eventually allows host selection [24]. Most changes on host plant use and preference are still consequence of modifications in gene expression patterns [25]. Phenotypic responses are affected by how or which genes are expressed, with the expectation that these variations occur in a complex way, with clusters of genes exerting influence on the expression of different genes, forming a large gene interaction network. ...
A dataset of gene expression from Spodoptera frugiperda, a highly generalist pest moth, was used to understand how gene regulation is related to larval host plant preference. Transcriptomic data of corn and rice strains of S. frugiperda larvae, reared on different diets, were analysed with three different approaches of gene network inference, namely co-expression, weighted co-expression and Bayesian networks, since each methodology provides a different visualization of the data. Using these approaches, it was possible to identify two loosely interconnected co-expression networks, one of them responsible for fast response to herbivory and anti-herbivory mechanisms and the other related to housekeeping genes, which present slower response to environmental variations. Integrating different levels of information such as gene expression patterns, gene assembly, transcriptomics, relationship among genes and phenotypes, functional relationships, among other information, enabled a wider visualization of S. frugiperda response to diet stimuli. The biological properties in the proposed networks are here described and discussed, as well as patterns of gene expression related to larval performance attributes.
... MYB8 is activated in response to JA-mediated induction by M. sexta larval herbivory (Onkokesung et al., 2012). However, JA levels in egg-primed, feeding-induced plants were not higher than in non-primed, feeding-induced plants when measured 1 day after wounding (Bandoly et al., 2015;Drok et al., 2018). ...
Plants can respond to eggs laid by herbivorous insects on their leaves by preparing (priming) their defense against the hatching larvae. Egg-mediated priming of defense is known for several plant species, including Brassicaceae. However, it is unknown yet for how long the eggs need to remain on a plant until a primed defense state is reached, which is ecologically manifested by reduced performance of the hatching larvae. To address this question, we used Arabidopsis thaliana, which carried eggs of the butterfly Pieris brassicae for 1–6 days prior to exposure to larval feeding. Our results show that larvae gained less biomass the longer the eggs had previously been on the plant. The strongest priming effect was obtained when eggs had been on the plant for 5 or 6 days, i.e., for (almost) the entire development time of the Pieris embryo inside the egg until larval hatching. Transcript levels of priming-responsive genes, levels of jasmonic acid-isoleucine (JA-Ile), and of the egg-inducible phytoalexin camalexin increased with the egg exposure time. Larval performance studies on mutant plants revealed that camalexin is dispensable for anti-herbivore defense against P. brassicae larvae, whereas JA-Ile – in concert with egg-induced salicylic acid (SA) – seems to be important for signaling egg-mediated primed defense. Thus, A. thaliana adjusts the kinetics of its egg-primed response to the time point of larval hatching. Hence, the plant is optimally prepared just in time prior to larval hatching.
... www.nature.com/scientificreports/ affect the feeding-induced changes of JA and SA levels one day after the onset of feeding by conspecific larvae [19][20][21][22] . Prior oviposition on N. attenuata did, however, result in a stronger transcriptional induction of the JA-responsive transcription factor MYB8 after feeding by larvae of the moths Spodoptera exigua or Manduca sexta 19,20 . ...
... We used standardised Generally Applicable Gene set Enrichment (GAGE) analyses 23 on gene ontology (GO) terms to investigate which plant biological processes (BPs) are mainly affected in response to (i) insect eggs, (ii) feeding by neonate larvae, and (iii) insect eggs followed by larval feeding (Fig. 1a). For the analysis we used transcriptomic data from published experiments with A. thaliana infested by P. brassicae 17 , N. attenuata infested by M. sexta or by S. exigua 21 , S. dulcamara infested by S. exigua 12,22 and U. minor infested by the leaf beetle Xanthogaleruca luteola 24 . Thus, our analysis is based on transcriptomic data obtained by studies of different plant-insect combinations, including plant species with very different life strategies, as well as lepidopteran and coleopteran insect species. ...
... the plant's transcriptional response to eggs, larval feeding and to the combination of eggs followed by larval feeding involves several phytohormone pathways. Our analysis, and the earlier original publications 12,17,21,22,24 to which our analysis refers, found prominent regulation of GS related to phytohormone signalling. Therefore, we compared the enrichment of all GS associated with phytohormone signalling and metabolism that were up-or down-regulated in at least one of the species combinations (Fig. 3, Supplementary Table S3). ...
Abstract Plants respond to insect eggs with transcriptional changes, resulting in enhanced defence against hatching larvae. However, it is unknown whether phylogenetically distant plant species show conserved transcriptomic responses to insect eggs and subsequent larval feeding. We used Generally Applicable Gene set Enrichment (GAGE) on gene ontology terms to answer this question and analysed transcriptome data from Arabidopsis thaliana, wild tobacco (Nicotiana attenuata), bittersweet nightshade (Solanum dulcamara) and elm trees (Ulmus minor) infested by different insect species. The different plant–insect species combinations showed considerable overlap in their transcriptomic responses to both eggs and larval feeding. Within these conformable responses across the plant–insect combinations, the responses to eggs and feeding were largely analogous, and about one-fifth of these analogous responses were further enhanced when egg deposition preceded larval feeding. This conserved transcriptomic response to eggs and larval feeding comprised gene sets related to several phytohormones and to the phenylpropanoid biosynthesis pathway, of which specific branches were activated in different plant–insect combinations. Since insect eggs and larval feeding activate conserved sets of biological processes in different plant species, we conclude that plants with different lifestyles share common transcriptomic alarm responses to insect eggs, which likely enhance their defence against hatching larvae.
... Previous studies have reported the induction of defense priming in response to different types of priming cues directly associated with herbivory. This includes priming by HIPVs (Engelberth et al., 2004;Paschold et al., 2006), oviposition-associated elicitors deposited on the oviposition site Drok, Bandoly, Stelzer, Lortzing, & Steppuhn, 2018;Pashalidou, Lucas-Barbosa, van Loon, Dicke, & Fatouros, 2013), and herbivore-derived olfactory cues such as sex attractants (Helms, De Moraes, Tooker, & Mescher, 2013). ...
... affecting the chewing herbivore Trirhabda virgata, thus suggesting nonspecific defense priming in this system. On the other hand, there are studies documenting herbivore-specific induction of defense priming for volatiles or oviposition-mediated priming of plants (Bandoly, Grichnik, Hilker, & Steppuhn, 2016;Drok et al., 2018;Moreira et al., 2018b). In Baccharis salicifolia plants, the reproductive rate of either Uroleucon macolai (specialist aphid) or Aphis gossypii (generalist aphid) was only significantly affected or reduced when emitter and receiver plants were damaged by the same aphid species, thus suggesting herbivore-specific direct defense priming in receiver plants (Moreira et al., 2018b). ...
... In Baccharis salicifolia plants, the reproductive rate of either Uroleucon macolai (specialist aphid) or Aphis gossypii (generalist aphid) was only significantly affected or reduced when emitter and receiver plants were damaged by the same aphid species, thus suggesting herbivore-specific direct defense priming in receiver plants (Moreira et al., 2018b). Drok et al. (2018) found that oviposition by generalist Spodoptera exigua and specialist M. sexta primed N. attenuata plants in a species-specific manner for induced transcriptional and phytohormone response after larval feeding. In addition, oviposition itself primed N. attenuata plants for subsequent caterpillar attack. ...
Plants produce species‐specific herbivore‐induced plant volatiles (HIPVs) after damage. We tested the hypothesis that herbivore‐specific HIPVs prime neighboring plants to induce defenses specific to the priming herbivore. Since Manduca sexta (specialist) and Heliothis virescens (generalist) herbivory induced unique HIPV profiles in Nicotiana benthamiana, we used these HIPVs to prime receiver plants for defense responses to simulated herbivory (mechanical wounding and herbivore regurgitant application). Jasmonic acid (JA) accumulations and emitted volatile profiles were monitored as representative defense responses since JA is the major plant hormone involved in wound and defense signaling and HIPVs have been implicated as signals in tritrophic interactions. Herbivore species‐specific HIPVs primed neighboring plants, which produced 2 to 4 times more volatiles and JA after simulated herbivory when compared to similarly treated constitutive volatile‐exposed plants. However, HIPV‐exposed plants accumulated similar amounts of volatiles and JA independent of the combination of priming or challenging herbivore. Furthermore, volatile profiles emitted by primed plants depended only on the challenging herbivore species but not on the species‐specific HIPV profile of damaged emitter plants. This suggests that feeding by either herbivore species primed neighboring plants for increased HIPV emissions specific to the subsequently attacking herbivore and is probably controlled by JA.
