NaMYC2 transcription factor regulates a subset of plant defense responses in Nicotiana attenuata
© Woldemariam et al.; licensee BioMed Central Ltd. 2013
Received: 29 January 2013
Accepted: 25 April 2013
Published: 1 May 2013
To survive herbivore attack, plants have evolved potent mechanisms of mechanical or chemical defense that are either constitutively present or inducible after herbivore attack. Due to the costs of defense deployment, plants often regulate their biosynthesis using various transcription factors (TFs). MYC2 regulators belong to the bHLH family of transcription factors that are involved in many aspects of plant defense and development. In this study, we identified a novel MYC2 TF from N. attenuata and characterized its regulatory function using a combination of molecular, analytic and ecological methods.
The transcript and targeted metabolite analyses demonstrated that NaMYC2 is mainly involved in the regulation of the biosynthesis of nicotine and phenolamides in N. attenuata. In addition, using broadly-targeted metabolite analysis, we identified a number of other metabolite features that were regulated by NaMYC2, which, after full annotation, are expected to broaden our understanding of plant defense regulation. Unlike previous reports, the biosynthesis of jasmonates and some JA-/NaCOI1-dependent metabolites (e.g. HGL-DTGs) were not strongly regulated by NaMYC2, suggesting the involvement of other independent regulators. No significant differences were observed in the performance of M. sexta on MYC2-silenced plants, consistent with the well-known ability of this specialist insect to tolerate nicotine.
By regulating the biosynthesis of nicotine, NaMYC2 is likely to enhance plant resistance against non-adapted herbivores and contribute to plant fitness; however, multiple JA/NaCOI1-dependent mechanisms (perhaps involving other MYCs) that regulate separate defense responses are likely to exist in N. attenuata. The considerable variation observed amongst different plant families in the responses regulated by jasmonate signaling highlights the sophistication with which plants craft highly specific and fine-tuned responses against the herbivores that attack them.
In their natural habitats, plants are exposed to a number of abiotic (e.g. drought, ultra-violet radiation, salinity) and biotic (e.g. herbivore and/or pathogen attack, competition) stresses which strongly undermine their Darwinian fitness. To cope with herbivory, plants have evolved intricate defense mechanisms that include mechanical barriers, trichomes, thorns, latex, waxes, and a toxic-/anti-nutritive chemical arsenal deployed either constitutively (e.g. nicotine, glucosinolates) or following herbivore attack (e.g. hydroxygeranyllinalool-diterpene glycosides (HGL-DTGs), phenolamides, trypsin protease inhibitors) [1–3]. In addition, and in concert with these direct defenses, plants recruit predators or parasitoids of the attackers using informative volatile organic compounds or nutritional rewards [4–6]. However, the costs of defense responses [2, 7, 8] necessitate the development of stringent regulatory mechanisms and several families of plant transcription factors (TFs) (e.g. ERF, bZIP, MYB, bHLH and WRKY) have been shown to regulate plant defense against biotic and abiotic stresses [9–11]. Many of these transcription factors are co-induced in response to different stresses suggesting the existence of complex interaction [12–14].
In many plant species, the role of phytohormones in coordinating the development of defense responses has clearly been shown, frequently with cross-talk among them to achieve intricately fine-tuned response outcomes [15–18]. Specifically, the jasmonate signaling pathway plays a critical role in mediating defense responses against herbivores [19–21]. In response to herbivore attack, GLA1 enzymes release 18:3 α-linolenic acid (α-LeA) from chloroplast membranes. α-LeA is subsequently converted to oxophytodienoic acid (OPDA) in the chloroplasts by lipoxygenase (LOX), allene oxide synthase (AOS) and allene oxide cyclase (AOC) enzymes. OPDA is transported to peroxisomes and oxidized by OPDA reductase (OPR) forming jasmonic acid (JA). In the cytosol, JA is conjugated to isoleucine by JAR enzymes that produce the bioactive jasmonate, (+)-7-iso-jasmonoyl-L-isoleucine (JA-Ile) [22, 23]. JA-Ile associates with the SCFCOI1 complex, presumably to ubiquinate JAZ repressors and tag them for degradation by the 26S proteasome. In the absence of stressful conditions, MYC2 is repressed by the JAZ repressors, which recruit TOPLESS (TPL) as a co-repressor either directly through the EAR (Ethylene Response Factor-Associated Amphifilic Repression) motif or using the EAR motif of the NINJA (Novel Interactor of JAZ) protein [24, 25]. Degradation of JAZ proteins releases the MYC2 transcription factor from repression and reconfigures downstream transcriptional processes [11, 24, 26–28].
MYC2 is a member of the basic Helix-Loop-Helix (bHLH) family of transcription factors (TFs) [29, 30] that are characterized by structurally and functionally conserved domains in many plant species. One of these conserved domains, the basic (b) region, is used to bind to variants of the G-box hexamer (5'-CACNTG-3') found on the promoters of MYC2-regulated genes. The HLH and ZIP domains are used for homo-/hetero-dimerization, while the JID (JAZ Interacting Domain) domain is used to interact with JAZ proteins [11, 28, 29, 31–34].
MYC2 transcription factors participate in the regulation of many JA-dependent physiological processes: defense against herbivores/pathogens, drought tolerance, circadian clock, light signaling and root growth [11, 35–39]. Guo et al. , in a proteomic study that involved mock- or MeJA-treated wild type and myc2 plants, recently identified 27 differentially regulated, JA-inducible and MYC2 dependent proteins involved in glucosinolate metabolism (22%), stress and defense (33%), photosynthesis (22.2%), carbohydrate metabolism (7.4%), protein folding and degradation (11.1%), highlighting the very diverse roles of MYC2.
N. attenuata is a wild tobacco species native to the Great Basin Desert in Utah (USA) which our group has developed into an ecological plant model. The defense responses of this species against its specialist herbivore, Manduca sexta, are well studied, and include the production of potent secondary metabolites: nicotine, HGL-DTGs, phenolamides and protease inhibitors [10, 41–47]. In this study, we identified a putative MYC2 transcription factor in N. attenuata (NaMYC2) and characterized its role in defense response regulation using reverse genetic, transcriptomic and untargeted/targeted metabolomic approaches. Our transcriptomic and metabolomic data indicate a strong involvement of NaMYC2 in nicotine accumulation. However, silencing this gene had only a limited effect on the accumulation of other plant defense metabolites which strongly implicates the involvement of multiple independent and/or redundant transcriptional regulators in defense signaling of N. attenuata plants.
Results and discussion
NaMYC2 transcripts are induced after herbivory
Targeted analysis of secondary metabolite accumulation in MYC2-VIGS plants
Nicotine, phenolamides, hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) and phenolic compounds are among the potent, JA-dependent anti-herbivore compounds in N. attenuata[2, 10, 54, 55]. Their JA-dependent pattern of accumulation suggests that the biosynthesis of these compounds might be regulated by NaMYC2. To test this hypothesis, we used the MYC2-VIGS plants: previously, Saedler and Badwin  demonstrated that VIGS effectively knocks-down the expression of plant defense genes (e.g. PMT) in both leaves and roots of N. attenuata plants. Then, we used a targeted metabolomic approach to compare the accumulation of defensive secondary metabolites in untreated control and WOS-treated (24, 48 and 72 h) EV and MYC2-VIGS plants.
Overall, our results are consistent with the previous reports which demonstrated regulation of jasmonate-induced nicotine/alkaloid biosynthesis by MYC2 TFs. In N. tabacum Bright Yellow (BY-2) cells that were transformed with an inverted-repeat (ir)NtMYC2a/2b construct, the accumulations of nicotine and anatabine were significantly reduced compared to untransformed controls . The NtMYC2 protein was also shown to regulate nicotine biosynthesis either by directly binding to the promoters of nicotine biosynthetic genes in roots or activating NtERF189 which, in turn, activates genes involved in nicotine biosynthesis . In N. benthamiana, VIGS of two bHLH transcription factors (named NbbHLH1 and NbbHLH2) as well as NbERF1 and NbHB1 decreased MeJA-induced accumulation of nicotine . These results demonstrate both the regulatory functions of MYC2 and the involvement of a network of transcription factors in the regulation of nicotine biosynthesis. However, the functions of the tobacco MYC2 genes were not examined in the context of natural herbivore feeding; neither were the effects of these MYC2 genes on the accumulations of other tobacco defense metabolites (e.g. phenolamides, HGL-DTGs, etc.) studied. From the phylogenetic relationship of MYC/bHLH TFs in N. attenuata, N. tabacum and N. benthamiana (Figure 1) and our results, the presence of additional MYC TFs in N. attenuata is a reasonable prediction. Further characterization of these putative TFs might help to fully understand the biosynthesis and ecological consequences of nicotine/alkaloid biosynthesis. Moreover, characterization of additional regulators would complement the partial regulatory function of NaMYC2 in the control of different classes of N. attenuata defense metabolites, as demonstrated in the next sections.
Total hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) and TPI levels
Taken together and considering the JA-/COI1-dependence of HGL-DTG and TPI accumulation in N. attenuata, the biosynthesis of HGL-DTGs and TPIs in N. attenuata is likely regulated by a JA-dependent, but NaMYC2-independent mechanism. Alternatively, the function and/or synergism of an independent MYC2 gene in N. attenuata can explain the partial function of NaMYC2. In addition, similar to phenolamides, the accumulation of HGL-DTG and TPI is also strongly influenced by the developmental stage of the plants . Van Dam et al.  showed that the de novo synthesis of PIs is limited to the early stages of plant development and that flowering plants treated with methyl jasmonate did not significantly increase their local or systemic PI activity levels. In addition, damage to older leaves elicited a much weaker systemic response in younger leaves compared to younger source leaves, a pattern also reported from other studies in N. tabacum. Heiling et al. (2010) demonstrated that the concentrations of 17-hydroxygeranyllinalool diterpene glycosides (DTGs) were highest in most valuable young and reproductive tissues, which is required for effective defense of these tissues against herbivores in N. attenuata.
NaMYC2 and regulation of herbivory-induced phytohormone accumulation
Performance of the specialist herbivore on MYC2-VIGS plants
Large scale transcriptomic and metabolomic analysis of MYC2-silenced leaves
The role of MYC2 TFs in orchestrating plant defense and developmental processes in several plant species were previously reviewed [35, 68, 69]. As master regulators, MYC2 TFs may either directly regulate the genes responsible for defense metabolite biosynthesis or regulate their regulators [11, 68]. To provide information for further work, we used unbiased approaches and compared herbivore-induced (WOS) changes in the transcriptome and metabolome of EV and MYC2-VIGS N. attenuata plants.
NaMYC2 regulated transcriptome ofN. attenuata
In contrast to independently performed qRT-PCR measurement of transcript abundances of phenolamide biosynthetic genes, the microarray analysis did not identify these genes (PAL, AT1, DH29 and MYB8) as differentially regulated in MYC2-VIGS plants compared to EV-VIGS plants because these genes did not pass the strict statistical criteria set for selection of at least 2-fold down-regulated genes in microarray experiment. Nicotine biosynthesis genes are only expressed in the roots and therefore could not be evaluated in the leaf samples used for microarrays.
Silencing of NaMYC2 significantly affects the N. attenuata metabolome
In many plant species, attack from herbivores elicits a cascade of complex transcriptional and metabolic responses that improve plant defense. The effectiveness of plant defense depends on the efficiency by which the timing and duration of responses are regulated. In this study, we identified a MYC2 TF in N. attenuata and characterized its regulatory role using transcriptomic and metabolomic approaches. Transcriptionally, we showed that the expressions of many genes, including transcription factors, involved in plant development or defense responses were affected when MYC2 was silenced in N. attenuata. This was supported by the metabolomic data which identified a large number of differentially regulated molecular features following the silencing. Most importantly, as was previously reported in N. tabacum and N. benthamiana, we showed that NaMYC2 regulates the in planta accumulation of nicotine in N. attenuata leaves. The fact that MYC2 did not strongly affect the accumulation of other JA-dependent metabolites, HGL-DTGs and proteinase inhibitors, suggests that another MYC TF is likely involved in the process.
Despite the considerable conservation of the basic components of plant defense responses among different plant species, substantial variations exist in the responses outcomes which highlights between-species differences in downstream regulatory fine-tuning [31, 73]. For example, in contrast to the considerable similarity among members of the genus Nicotiana in the regulation of nicotine biosynthesis by MYC2 [52, 58, 59] (Figure 1), silencing MYC2 in N. attenuata did not have the exact same effects as reported in A. thaliana; we did not observe a role of MYC2 either in a positive feedback loop activating JA biosynthesis or in a negative feedback involving suppression of the jasmonate response through the activation of JAZ repressors [11, 74].
In addition, not all JA-dependent defense metabolites (e.g. HGL-DTGs) were regulated by MYC2 in N. attenuata. In fact, when compared against the diversity of defense metabolites in N. attenuata, the regulatory function of MYC2 is quite limited. This rather limited role suggests that other members of the bHLH family of transcription factors might be involved in the regulation of defense responses not regulated by MYC2. The recent identification of additional MYC2 TFs in A. thaliana[36, 37], N. tabacum[52, 58] and N. benthamiana with overlapping or distinct functions support this conjecture.
Indeed, we found an additional MYC2-like gene (KC906192) in diploid N. attenuata showing a 72.3% protein sequence identity with NaMYC2. In the phylogenetic analysis, MYC2-like protein clustered separately from the MYC2 clade of Solanaceae species, including N. tabacum MYC2a and MYC2b. When we briefly examined the function of MYC2-like gene in N. attenuata, interestingly, increased defense responses in MYC2-like-VIGS plants were observed (data not shown). This was in a strong contrast to silencing the NaMYC2 (and N. benthamiana genes bHLH1and bHLH2; ) but in agreement with the VIGS–induced silencing of the N. benthamiana bHLH3 (a gene fragment not included in phylogenetic tree shown in Figure 1), which increased the nicotine content in the VIGS-silenced N. benthamiana plants after foliar application of MeJA . Therefore, some of the MYC2-like genes may work as repressors of JA-induced responses, contributing to a fine-tuning of defense against herbivores, possibly by competing for promoter binding sites with the activator-type MYC2 genes. As previously demonstrated for the transient character of JA-Ile accumulation [62, 75], tight control of JA signaling is likely to be essential for plant responses to multiple biotic stresses in the environment. Identification and characterization of additional MYC2 TFs in N. attenuata and other plant species is likely to provide a more complete mechanistic picture of JA-regulated defense responses.
Considering the high degree of conservation in the binding site of MYC2 TFs in different species [29, 31], we believe future research in determining the binding sites of these TFs will be critical to understanding their function. When these binding sites are identified, additional MYC2-dependent genes or other transcription factors that respond to herbivory, disease, environmental stress or development can be more readily identified. It would be interesting to identify the interacting partners of MYC2 TFs in N. attenuata and characterize the mechanisms of interaction to understand how the signaling components evolved. In A. thaliana, transcriptional regulation by MYC2 requires interactions with important regulatory elements including members of the mediator complex proteins (e.g. MED25), chromatin-opening proteins like General Control Non-repressible 5 (GCN5), members of the histone acetyl transferase family and SPLAYED (SYD) [35, 76, 77]. Identification and characterization of homologues of these components in N. attenuata might test the generality of the signaling processes across different plant families.
Plant growth and treatments
N. attenuata seeds that were collected from its native habitat in Great Basin desert, Utah (USA) and inbred for 31 generations were used for the experiments. Seed germination and plant growth conditions were described in Krügel et al. . To experimentally simulate herbivory, we wounded fully expanded leaves of EV and MYC2-VIGS (n=5) N. attenuata plants with a serrated fabric pattern wheel and the wounds were treated with 20 μL of diluted (1:5, v/v in water) M. sexta oral secretions (WOS), while controls were collected from untreated plants. To evaluate performance of the specialist herbivore (M. sexta) on transformed plants, freshly hatched neonates were fed on EV and transformed plants (n = 20) and their masses were measured every 4 d.
Virus Induced Gene Silencing (VIGS)
Virus Induced Gene Silencing (VIGS) system, described in Saedler and Baldwin , was used to transiently silence MYC2 transcription factor. Briefly, we amplified ~250 bp fragment of the N. attenuata MYC2 using specific primers (Additional file 4: Table S3), cloned them into the PTV00 vector. We verified the clone by sequencing and transformed GV3101 strain of Agrobacterium tumefaciens with either untransformed plasmid (PTV00, control) or plasmids harboring the inserts (pTV-MYC2) and incubated them at 26°C for two days. On the day of infiltration, overnight cultures of all constructs and pBINTRA and pTVPDS were inoculated into YEP media containing antibiotics (Kanamycin 50 mg/L) and incubated (28°C) for 5 h. When the cultures attained an OD of 0.6 to 0.8, we centrifuged them (1,125g, 4°C for 5 min), resuspended the pellets in an equimolar mix (5 mM) of MgCl2 and MES and prepared a 1:1 mix of each construct with the helper strain pBINTRA. Using 1 mL syringes, we infiltrated the suspension into five leaves of 25 d old N. attenuata plants, covered them with plastic and left them in a dark chamber for 2 d. The plants were kept in the growth chamber under 16 h/day, 8 h/night light regime at 22°C. We monitored the spread of silencing using control plants infiltrated with the pTVPDS construct which induced leaf bleaching, while the efficiency of silencing was determined by measuring transcript abundances using qRT-PCR.
We treated fully elongated leaves of EV and MYC2-VIGS plants (n = 3) with WOS for 1h, collected and ground the leaves in liquid nitrogen and extracted RNA for the microarray analysis as described in Gillardoni et al. . After hybridization and array processing, we normalized (with the 75th percentile of the respective columns) and log2-transformed the raw expression values obtained from the "gProcessedSignal" column and processed them using Significance of Microarrays (SAM; http://www-stat.stanford.edu/~tibs/SAM/) package on Excel (Microsoft). For the analysis, we set the minimum fold change, delta and median FDR (%) values to 2, 0.69 and 15.8 (%) respectively. Genes that differed significantly in comparison to EV plants were annotated using Blast2Go  and grouped according to TAIR classification. The microarray data was deposited in GEO under the accession number GSE45608.
Transcript abundance measurement
We extracted total RNA from frozen leaf material of untreated or WOS-treated EV and MYC2-VIGS plants (n = 5) using TRIzol reagent (Invitrogen) as recommended by the manufacturer. We treated the total RNA with DNAse (RQ1 RNase-Free DNase; Promega) before synthesizing cDNA using oligo (dT)18 and Superscript II reverse transcriptase (Invitrogen). Transcript abundances were measured on Mx3005P Multiplex qPCR (Stratagene) with qPCR core kit for SYBR Green I (Eurogentec). Relative transcript abundances were determined by comparing sample fluorescence signals to dilution series of cDNA prepared from the 1 h WOS -treated samples, and examined on the same plate. Signals were then normalized by the average EF-1α transcript abundances determined separately for each sample. The primers used for qRT-PCR are listed in Additional file 4: Table S3. As there is a considerable similarity in protein coding sequences in multiple members in bHLH TF family, it may imply significant functional redundancy of these regulators in biological systems. We therefore carefully designed our primers in the 3’ non-translated end of the gene to amplify and detect specifically the NaMYC2 transcription factor (Additional file 1: Figure S2).
Fully-expanded leaves of EV and MYC2-VIGS plants (n = 5) were treated with WOS for 1 h or 2 h, collected and ground in liquid nitrogen and stored at −80°C until use. We homogenized about 200 mg powder in 1 mL ethyl acetate (containing 200 ng/mL D2-JA and 40 ng/mL D6-ABA, D4-SA and JA-13C6-Ile internal standards), centrifuged for 20 min (16,100g, 4°C) and transferred the supernatants into new tubes. After re-extracting the pellets with 0.5 mL ethyl acetate and combining the supernatants, we evaporated the ethyl acetate on a vacuum concentrator (Eppendorf) and resuspended the residue in 0.5 mL 70% methanol in water (v/v). Then, we centrifuged the re-suspended samples for 10 min (16,100 g, 4°C) and analyzed the supernatant (10 μL) on Varian 1200L Triple-Quadrupole-LC-MS (Varian) using a ProntoSIL® column (C18; 5 μm, 50 × 2 mm; Bischoff) attached to a precolumn (C18; 4 × 2 mm, Phenomenex). Detail measurement conditions are described in Woldemariam et al. .
Secondary metabolite analysis
To undertake targeted defense secondary metabolite (nicotine, total 17-hydroxygeranyllinalool diterpene glycosides [HGL-DTGs], caffeoylputrescine, dicaffeoylspermidine, chlorogenic acid and rutin) analysis, we treated leaves of EV and MYC2-VIGS (n = 5) plants with WOS for 24, 48 or 72 h, collected and ground the samples in liquid nitrogen. Control samples were collected without treatment. About 100 mg powder was extracted and analyzed on HPLC equipped with a photodiode array detector as previously described in Onkokesung et al. .
Untargeted metabolomic analysis
To undertake an unbiased metabolomic analysis, metabolites were extracted from leaves (n = 3) of EV and MYC2 silenced N. attenuata plants fed on for 4 d by neonates of M. sexta and analyzed on an HPLC 1100 Series system (Agilent, Palo Alto, USA) coupled to a MicroToF mass spectrometer (Bruker Daltonik, Bremen, Germany). The optimized analytic procedures are described in Gaquerel et al. . Briefly, peak picking, peak detection and RT corrections were performed by XCMS (and CAMERA) package using the following parameters: centWave method; ppm = 20; snthresh =10; peakwidth = between 5 and 18 s; minfrac=0.5; minsamp=1; bw=10; mzwid=0.01; sleep=0.001. To fill missing features, we used the FillPeaks function from XCMS. We exported the pre-processed data to Excel, filtered those features with RTs < 60 seconds and m/z < 80 and analyzed the processed data on Metaboanalyst 2.0 following the procedure described before .
We used STATVIEW (version 5.0; SAS Institute, Cary, NC, USA) software to perform statistical analyses with alpha level of 0.05 for all statistical tests.
We acknowledge the German Academic Exchange Service (DAAD) and the International Max Planck Research School (IMPRS) for financial support. Son Truong Dinh was also supported by the Vietnam Ministry of Agricultural and Rural Development and the Max Planck Society.
NaMYC2 GenBank Accession number: KC832837
NaMYC2-like GenBank Accession number: KC906192
- Wittstock U, Gershenzon J: Constitutive plant toxins and their role in defense against herbivores and pathogens. Curr Opin Plant Biol. 2002, 5 (4): 300-307.PubMedView ArticleGoogle Scholar
- Kessler A, Baldwin IT: Plant responses to insect herbivory: the emerging molecular analysis. Annu Rev Plant Biol. 2002, 53: 299-328.PubMedView ArticleGoogle Scholar
- Wu JQ, Baldwin IT: New insights into plant responses to the attack from insect herbivores. Annu Rev Genet. 2010, 44: 1-24.PubMedView ArticleGoogle Scholar
- Heil M: Plastic defence expression in plants. Evol Ecol. 2010, 24 (3): 555-569.View ArticleGoogle Scholar
- Mithofer A, Boland W: Plant defense against herbivores: chemical aspects. Annu Rev Plant Biol. 2012, 63: 431-450.PubMedView ArticleGoogle Scholar
- Baldwin IT: Plant volatiles. Curr Biol. 2010, 20 (9): 392-397.View ArticleGoogle Scholar
- Baldwin IT: Jasmonate-induced responses are costly but benefit plants under attack in native populations. Proc Natl Acad Sci U S A. 1998, 95 (14): 8113-8118.PubMedPubMed CentralView ArticleGoogle Scholar
- Heil M, Baldwin IT: Fitness costs of induced resistance: emerging experimental support for a slippery concept. Trends Plant Sci. 2002, 7 (2): 61-67.PubMedView ArticleGoogle Scholar
- Gigolashvili T, Yatusevich R, Berger B, Muller C, Flugge UI: The R2R3-MYB transcription factor HAG1/MYB28 is a regulator of methionine-derived glucosinolate biosynthesis in Arabidopsis thaliana. Plant J. 2007, 51 (2): 247-261.PubMedView ArticleGoogle Scholar
- Kaur H, Heinzel N, Schottner M, Baldwin IT, Galis I: R2R3-NaMYB8 regulates the accumulation of phenylpropanoid-polyamine conjugates, which are essential for local and systemic defense against insect herbivores in Nicotiana attenuata. Plant Physiol. 2010, 152 (3): 1731-1747.PubMedPubMed CentralView ArticleGoogle Scholar
- Dombrecht B, Xue GP, Sprague SJ, Kirkegaard JA, Ross JJ, Reid JB, Fitt GP, Sewelam N, Schenk PM, Manners JM: MYC2 differentially modulates diverse jasmonate-dependent functions in Arabidopsis. Plant Cell. 2007, 19 (7): 2225-2245.PubMedPubMed CentralView ArticleGoogle Scholar
- Singh KB, Foley RC, Onate-Sanchez L: Transcription factors in plant defense and stress responses. Curr Opin Plant Biol. 2002, 5 (5): 430-436.PubMedView ArticleGoogle Scholar
- Endt DV, Kijne JW, Memelink J: Transcription factors controlling plant secondary metabolism: what regulates the regulators?. Phytochemistry. 2002, 61 (2): 107-114.View ArticleGoogle Scholar
- De Boer K, Tilleman S, Pauwels L, Vanden Bossche R, De Sutter V, Vanderhaeghen R, Hilson P, Hamill JD, Goossens A: APETALA2/ETHYLENE RESPONSE FACTOR and basic helix-loop-helix tobacco transcription factors cooperatively mediate jasmonate-elicited nicotine biosynthesis. Plant J. 2011, 66 (6): 1053-1065.PubMedView ArticleGoogle Scholar
- Arimura GI, Ozawa R, Maffei ME: Recent advances in plant early signaling in response to herbivory. Int J Mol Sci. 2011, 12 (6): 3723-3739.PubMedPubMed CentralView ArticleGoogle Scholar
- Verhage A, van Wees SC, Pieterse CM: Plant immunity: it's the hormones talking, but what do they say?. Plant Physiol. 2010, 154 (2): 536-540.PubMedPubMed CentralView ArticleGoogle Scholar
- Bari R, Jones J: Role of plant hormones in plant defence responses. Plant Mol Biol. 2009, 69 (4): 473-488.PubMedView ArticleGoogle Scholar
- Grunewald W, Vanholme B, Pauwels L, Plovie E, Inze D, Gheysen G, Goossens A: Expression of the Arabidopsis jasmonate signalling repressor JAZ1/TIFY10A is stimulated by auxin. Embo Rep. 2009, 10 (8): 923-928.PubMedPubMed CentralView ArticleGoogle Scholar
- Hause B, Wasternack C, Strack D: Jasmonates in stress responses and development. Phytochemistry. 2009, 70 (13–14): 1483-1484.PubMedView ArticleGoogle Scholar
- Koo AJK, Howe GA: The wound hormone jasmonate. Phytochemistry. 2009, 70 (13–14): 1571-1580.PubMedPubMed CentralView ArticleGoogle Scholar
- Abe H, Shimoda T, Ohnishi J, Kugimiya S, Narusaka M, Seo S, Narusaka Y, Tsuda S, Kobayashi M: Jasmonate-dependent plant defense restricts thrips performance and preference. BMC Plant Biol. 2009, 9: 97.PubMedPubMed CentralView ArticleGoogle Scholar
- Schaller A, Stintzi A: Enzymes in jasmonate biosynthesis - Structure, function, regulation. Phytochemistry. 2009, 70 (13–14): 1532-1538.PubMedView ArticleGoogle Scholar
- Fonseca S, Chini A, Hamberg M, Adie B, Porzel A, Kramell R, Miersch O, Wasternack C, Solano R: (+)-7-iso-Jasmonoyl-L-isoleucine is the endogenous bioactive jasmonate. Nat Chem Biol. 2009, 5 (5): 344-350.PubMedView ArticleGoogle Scholar
- Wasternack C, Kombrink E: Jasmonates: structural requirements for lipid-derived signals active in plant stress responses and development. Acs Chemical Biology. 2010, 5 (1): 63-77.PubMedView ArticleGoogle Scholar
- Pauwels L, Barbero GF, Geerinck J, Tilleman S, Grunewald W, Perez AC, Chico JM, Vanden Bossche R, Sewell J, Gil E: NINJA connects the co-repressor TOPLESS to jasmonate signalling. Nature. 2010, 464 (7289): 788-791.PubMedPubMed CentralView ArticleGoogle Scholar
- Katsir L, Chung HS, Koo AJK, Howe GA: Jasmonate signaling: a conserved mechanism of hormone sensing. Curr Opin Plant Biol. 2008, 11 (4): 428-435.PubMedPubMed CentralView ArticleGoogle Scholar
- Gfeller A, Liechti R, Farmer EE: Arabidopsis jasmonate signaling pathway. Sci Signal. 2010, 3 (109): cm4.PubMedGoogle Scholar
- Chini A, Fonseca S, Fernandez G, Adie B, Chico JM, Lorenzo O, Garcia-Casado G, Lopez-Vidriero I, Lozano FM, Ponce MR: The JAZ family of repressors is the missing link in jasmonate signalling. Nature. 2007, 448 (7154): 666-U664.PubMedView ArticleGoogle Scholar
- Carretero-Paulet L, Galstyan A, Roig-Villanova I, Martinez-Garcia JF, Bilbao-Castro JR, Robertson DL: Genome-wide classification and evolutionary analysis of the bHLH family of transcription factors in Arabidopsis, poplar, rice, moss, and algae. Plant Physiol. 2010, 153 (3): 1398-1412.PubMedPubMed CentralView ArticleGoogle Scholar
- Heim MA, Jakoby M, Werber M, Martin C, Weisshaar B, Bailey PC: The basic helix-loop-helix transcription factor family in plants: a genome-wide study of protein structure and functional diversity. Mol Biol Evol. 2003, 20 (5): 735-747.PubMedView ArticleGoogle Scholar
- Boter M, Ruiz-Rivero O, Abdeen A, Prat S: Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis. Genes Dev. 2004, 18 (13): 1577-1591.PubMedPubMed CentralView ArticleGoogle Scholar
- Yadav V, Mallappa C, Gangappa SN, Bhatia S, Chattopadhyay S: A basic helix-loop-helix transcription factor in Arabidopsis, MYC2, acts as a repressor of blue light-mediated photomorphogenic growth. Plant Cell. 2005, 17 (7): 1953-1966.PubMedPubMed CentralView ArticleGoogle Scholar
- Figueroa P, Browse J: The Arabidopsis JAZ2 promoter contains a G-box and thymidine-rich module that are necessary and sufficient for jasmonate-dependent activation by MYC transcription factors and repression by JAZ proteins. Plant Cell Physiol. 2012, 53 (2): 330-343.PubMedView ArticleGoogle Scholar
- Amoutzias GD, Robertson DL, de Peer YV, Oliver SG: Choose your partners: dimerization in eukaryotic transcription factors. Trends Biochem Sci. 2008, 33 (5): 220-229.PubMedView ArticleGoogle Scholar
- Kazan K, Manners JM: MYC2: the master in action. Mol Plant. 2012, 10.1093/mp/sss128.Google Scholar
- Niu YJ, Figueroa P, Browse J: Characterization of JAZ-interacting bHLH transcription factors that regulate jasmonate responses in Arabidopsis. J Exp Bot. 2011, 62 (6): 2143-2154.PubMedPubMed CentralView ArticleGoogle Scholar
- Fernandez-Calvo P, Chini A, Fernandez-Barbero G, Chico JM, Gimenez-Ibanez S, Geerinck J, Eeckhout D, Schweizer F, Godoy M, Franco-Zorrilla JM: The Arabidopsis bHLH transcription factors MYC3 and MYC4 are targets of JAZ repressors and act additively with MYC2 in the activation of jasmonate responses. Plant Cell. 2011, 23 (2): 701-715.PubMedPubMed CentralView ArticleGoogle Scholar
- Lorenzo O, Chico JM, Sanchez-Serrano JJ, Solano R: Jasmonate-insensitive1 encodes a MYC transcription factor essential to discriminate between different jasmonate-regulated defense responses in Arabidopsis. Plant Cell. 2004, 16 (7): 1938-1950.PubMedPubMed CentralView ArticleGoogle Scholar
- Verhage A, Vlaardingerbroek I, Raaymakers C, Van Dam NM, Dicke M, Van Wees SC, Pieterse CM: Rewiring of the jasmonate signaling pathway in Arabidopsis during insect herbivory. Front Plant Sci. 2011, 2: 47.PubMedPubMed CentralView ArticleGoogle Scholar
- Guo J, Pang Q, Wang L, Yu P, Li N, Yan X: Proteomic identification of MYC2-dependent jasmonate-regulated proteins in Arabidopsis thaliana. Proteome Sci. 2012, 10 (1): 57.PubMedPubMed CentralView ArticleGoogle Scholar
- Halitschke R, Ziegler J, Keinanen M, Baldwin IT: Silencing of hydroperoxide lyase and allene oxide synthase reveals substrate and defense signaling crosstalk in Nicotiana attenuata. Plant J. 2004, 40 (1): 35-46.PubMedView ArticleGoogle Scholar
- Kallenbach M, Alagna F, Baldwin IT, Bonaventure G: Nicotiana attenuata SIPK, WIPK, NPR1, and fatty acid-amino acid conjugates participate in the induction of jasmonic acid biosynthesis by affecting early enzymatic steps in the pathway. Plant Physiol. 2010, 152 (1): 96-106.PubMedPubMed CentralView ArticleGoogle Scholar
- Meldau S, Baldwin IT, Wu J: SGT1 regulates wounding- and herbivory-induced jasmonic acid accumulation and Nicotiana attenuata's resistance to the specialist lepidopteran herbivore Manduca sexta. New Phytol. 2011, 189 (4): 1143-1156.PubMedView ArticleGoogle Scholar
- Skibbe M, Qu N, Galis I, Baldwin IT: Induced plant defenses in the natural environment: Nicotiana attenuata WRKY3 and WRKY6 coordinate responses to herbivory. Plant Cell. 2008, 20 (7): 1984-2000.PubMedPubMed CentralView ArticleGoogle Scholar
- Wang L, Allmann S, Wu J, Baldwin IT: Comparisons of LIPOXYGENASE3- and JASMONATE-RESISTANT4/6-silenced plants reveal that jasmonic acid and jasmonic acid-amino acid conjugates play different roles in herbivore resistance of Nicotiana attenuata. Plant Physiol. 2008, 146 (3): 904-915.PubMedPubMed CentralView ArticleGoogle Scholar
- Paschold A, Bonaventure G, Kant MR, Baldwin IT: Jasmonate perception regulates jasmonate biosynthesis and JA-Ile metabolism: the case of COI1 in Nicotiana attenuata. Plant Cell Physiol. 2008, 49 (8): 1165-1175.PubMedView ArticleGoogle Scholar
- Onkokesung N, Gaquerel E, Kotkar H, Kaur H, Baldwin IT, Galis I: MYB8 controls inducible phenolamide levels by activating three novel hydroxycinnamoyl-coenzyme A:polyamine transferases in Nicotiana attenuata. Plant Physiol. 2012, 158 (1): 389-407.PubMedPubMed CentralView ArticleGoogle Scholar
- Vogel H, Kroymann J, Mitchell-Olds T: Different transcript patterns in response to specialist and generalist herbivores in the wild Arabidopsis relative Boechera divaricarpa. PLoS One. 2007, 2 (10): e1081.PubMedPubMed CentralView ArticleGoogle Scholar
- Schmidt DD, Voelckel C, Hartl M, Schmidt S, Baldwin IT: Specificity in ecological interactions: attack from the same lepidopteran herbivore results in species-specific transcriptional responses in two solanaceous host plants. Plant Physiol. 2005, 138 (3): 1763-1773.PubMedPubMed CentralView ArticleGoogle Scholar
- Ehlting J, Chowrira SG, Mattheus N, Aeschliman DS, Arimura GI, Bohlmann J: Comparative transcriptome analysis of Arabidopsis thaliana infested by diamond back moth (Plutella xylostella) larvae reveals signatures of stress response, secondary metabolism, and signalling. BMC Genomics. 2008, 9: 154.PubMedPubMed CentralView ArticleGoogle Scholar
- Bodenhausen N, Reymond P: Signaling pathways controlling induced resistance to insect herbivores in Arabidopsis. Mol Plant Microbe Interact. 2007, 20 (11): 1406-1420.PubMedView ArticleGoogle Scholar
- Shoji T, Hashimoto T: Tobacco MYC2 regulates jasmonate-inducible nicotine biosynthesis genes directly and by way of the NIC2-locus ERF genes. Plant Cell Physiol. 2011, 52 (6): 1117-1130.PubMedView ArticleGoogle Scholar
- Kim SG, Yon F, Gaquerel E, Gulati J, Baldwin IT: Tissue specific diurnal rhythms of metabolites and their regulation during herbivore attack in a native tobacco. Nicotiana attenuata Plos One. 2011, 6 (10): e26214.PubMedView ArticleGoogle Scholar
- Kessler A, Halitschke R, Baldwin IT: Silencing the jasmonate cascade: induced plant defenses and insect populations. Science. 2004, 305 (5684): 665-668.PubMedView ArticleGoogle Scholar
- Heiling S, Schuman MC, Schoettner M, Mukerjee P, Berger B, Schneider B, Jassbi AR, Baldwin IT: Jasmonate and ppHsystemin regulate key malonylation steps in the biosynthesis of 17-hydroxygeranyllinalool diterpene glycosides, an abundant and effective direct defense against herbivores in Nicotiana attenuata. Plant Cell. 2010, 22 (1): 273-292.PubMedPubMed CentralView ArticleGoogle Scholar
- Saedler R, Baldwin IT: Virus-induced gene silencing of jasmonate-induced direct defences, nicotine and trypsin proteinase-inhibitors in Nicotiana attenuata. J Exp Bot. 2004, 55 (395): 151-157.PubMedView ArticleGoogle Scholar
- Steppuhn A, Gase K, Krock B, Halitschke R, Baldwin IT: Nicotine's defensive function in nature. PLoS Biol. 2004, 2 (8): E217.PubMedPubMed CentralView ArticleGoogle Scholar
- Zhang HB, Bokowiec MT, Rushton PJ, Han SC, Timko MP: Tobacco transcription factors NtMYC2a and NtMYC2b form nuclear complexes with the NtJAZ1 repressor and regulate multiple jasmonate-inducible steps in nicotine biosynthesis. Molecular Plant. 2012, 5 (1): 73-84.PubMedView ArticleGoogle Scholar
- Todd AT, Liu EW, Polvi SL, Pammett RT, Page JE: A functional genomics screen identifies diverse transcription factors that regulate alkaloid biosynthesis in Nicotiana benthamiana. Plant J. 2010, 62 (4): 589-600.PubMedView ArticleGoogle Scholar
- Jassbi AR, Gase K, Hettenhausen C, Schmidt A, Baldwin IT: Silencing geranylgeranyl diphosphate synthase in Nicotiana attenuata dramatically impairs resistance to tobacco hornworm. Plant Physiol. 2008, 146 (3): 974-986.PubMedPubMed CentralView ArticleGoogle Scholar
- Dinh ST, Galis I, Baldwin IT: UVB radiation and 17-hydroxygeranyllinalool diterpene glycosides provide durable resistance against mirid (Tupiocoris notatus) attack in field-grown Nicotiana attenuata plants. Plant Cell Environ. 2013, 36 (3): 590-606.View ArticleGoogle Scholar
- Woldemariam MG, Onkokesung N, Baldwin IT, Galis I: Jasmonoyl-L-isoleucine hydrolase 1 (JIH1) regulates jasmonoyl-L-isoleucine levels and attenuates plant defenses against herbivores. Plant J. 2012, 72 (5): 758-767.PubMedView ArticleGoogle Scholar
- Jongsma MA, Bakker PL, Visser B, Stiekema WJ: Trypsin inhibitor activity in mature tobacco and tomato plants is mainly induced locally in response to insect attack, wounding and virus infection. Planta. 1994, 195 (1): 29-35.View ArticleGoogle Scholar
- Paschold A, Halitschke R, Baldwin IT: Co(i)-ordinating defenses: NaCOI1 mediates herbivore-induced resistance in Nicotiana attenuata and reveals the role of herbivore movement in avoiding defenses. Plant J. 2007, 51 (1): 79-91.PubMedView ArticleGoogle Scholar
- Diezel C, Allmann S, Baldwin IT: Mechanisms of optimal defense patterns in Nicotiana attenuata: flowering attenuates herbivory-elicited ethylene and jasmonate signaling. J Integr Plant Biol. 2011, 53 (12): 971-983.PubMedView ArticleGoogle Scholar
- van Dam NM, Horn M, Mares M, Baldwin IT: Ontogeny constrains systemic protease inhibitor response in Nicotiana attenuata. J Chem Ecol. 2001, 27 (3): 547-568.PubMedView ArticleGoogle Scholar
- Pearce G, Johnson S, Ryan CA: Purification and characterization from tobacco (Nicotiana tabacum) leaves of six small, wound-inducible, proteinase isoinhibitors of the potato inhibitor-II family. Plant Physiol. 1993, 102 (2): 639-644.PubMedPubMed CentralView ArticleGoogle Scholar
- Woldemariam MG, Baldwin IT, Galis I: Transcriptional regulation of plant inducible defenses against herbivores: a mini-review. J Plant Interact. 2011, 6 (2–3): 113-119.View ArticleGoogle Scholar
- Yang CQ, Fang X, Wu XM, Mao YB, Wang LJ, Chen XY: Transcriptional regulation of plant secondary metabolism. J Integr Plant Biol. 2012, 54 (10): 703-712.PubMedView ArticleGoogle Scholar
- Cheng Z, Sun L, Qi T, Zhang B, Peng W, Liu Y, Xie D: The bHLH transcription factor MYC3 interacts with the jasmonate ZIM-domain proteins to mediate jasmonate response in Arabidopsis. Mol Plant. 2011, 4 (2): 279-288.PubMedView ArticleGoogle Scholar
- Gangappa SN, Prasad VB, Chattopadhyay S: Functional interconnection of MYC2 and SPA1 in the photomorphogenic seedling development of Arabidopsis. Plant Physiol. 2010, 154 (3): 1210-1219.PubMedPubMed CentralView ArticleGoogle Scholar
- Gaquerel E, Heiling S, Schoettner M, Zurek G, Baldwin IT: Development and validation of a liquid chromatography-electrospray ionization-time-of-flight mass spectrometry method for induced changes in Nicotiana attenuata leaves during simulated herbivory. J Agr Food Chem. 2010, 58 (17): 9418-9427.View ArticleGoogle Scholar
- VanDoorn A, Bonaventure G, Schmidt DD, Baldwin IT: Regulation of jasmonate metabolism and activation of systemic signaling in Solanum nigrum: COI1 and JAR4 play overlapping yet distinct roles. New Phytol. 2011, 190 (3): 640-652.PubMedView ArticleGoogle Scholar
- Oh Y, Baldwin IT, Galis I: NaJAZh regulates a subset of defense responses against herbivores and spontaneous leaf necrosis in Nicotiana attenuata plants. Plant Physiol. 2012, 159 (2): 769.PubMedPubMed CentralView ArticleGoogle Scholar
- Miersch O, Neumerkel J, Dippe M, Stenzel I, Wasternack C: Hydroxylated jasmonates are commonly occurring metabolites of jasmonic acid and contribute to a partial switch-off in jasmonate signaling. New Phytol. 2008, 177 (1): 114-127.PubMedGoogle Scholar
- Chen R, Jiang HL, Li L, Zhai QZ, Qi LL, Zhou WK, Liu XQ, Li HM, Zheng WG, Sun JQ: The Arabidopsis mediator subunit MED25 differentially regulates jasmonate and abscisic acid signaling through interacting with the MYC2 and ABI5 transcription factors. Plant Cell. 2012, 24 (7): 2898-2916.PubMedPubMed CentralView ArticleGoogle Scholar
- Wu K, Zhang L, Zhou C, Yu CW, Chaikam V: HDA6 is required for jasmonate response, senescence and flowering in Arabidopsis. J Exp Bot. 2008, 59 (2): 225-234.PubMedView ArticleGoogle Scholar
- Krugel T, Lim M, Gase K, Halitschke R, Baldwin IT: Agrobacterium-mediated transformation of Nicotiana attenuata, a model ecological expression system. Chemoecology. 2002, 12 (4): 177-183.View ArticleGoogle Scholar
- Gilardoni PA, Hettenhausen C, Baldwin IT, Bonaventure G: Nicotiana attenuata LECTIN RECEPTOR KINASE1 suppresses the insect-mediated inhibition of induced defense responses during Manduca sexta herbivory. Plant Cell. 2011, 23 (9): 3512-3532.PubMedPubMed CentralView ArticleGoogle Scholar
- Gotz S, Garcia-Gomez JM, Terol J, Williams TD, Nagaraj SH, Nueda MJ, Robles M, Talon M, Dopazo J, Conesa A: High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids Res. 2008, 36 (10): 3420-3435.PubMedPubMed CentralView ArticleGoogle Scholar
- Onkokesung N, Galis I, von Dahl CC, Matsuoka K, Saluz HP, Baldwin IT: Jasmonic acid and ethylene modulate local responses to wounding and simulated herbivory in Nicotiana attenuata leaves. Plant Physiol. 2010, 153 (2): 785-798.PubMedPubMed CentralView ArticleGoogle Scholar
- Xia JG, Mandal R, Sinelnikov IV, Broadhurst D, Wishart DS: MetaboAnalyst 2.0-a comprehensive server for metabolomic data analysis. Nucleic Acids Res. 2012, 40 (W1): W127-W133.PubMedPubMed CentralView ArticleGoogle Scholar
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