RESUMO
Bemisia tabaci (whitefly) is a polyphagous agroeconomic pest species complex. Two members of this species complex, Mediterranean (MED) and Middle-East-Asia Minor 1 (MEAM1), have a worldwide distribution and have been shown to manipulate plant defenses through effectors. In this study, we used three different strategies to identify three MEAM1 proteins that can act as effectors. Effector B1 was identified using a bioinformatics-driven effector-mining strategy, whereas effectors S1 and P1 were identified in the saliva of whiteflies collected from artificial diet and in phloem exudate of tomato on which nymphs were feeding, respectively. These three effectors were B. tabaci specific and able to increase whitefly fecundity when transiently expressed in tobacco plants (Nicotiana tabacum). Moreover, they reduced growth of Pseudomonas syringae pv. tabaci in Nicotiana benthamiana. All three effectors changed gene expression in planta, and B1 and S1 also changed phytohormone levels. Gene ontology and KEGG pathway enrichment analysis pinpointed plant-pathogen interaction and photosynthesis as the main enriched pathways for all three effectors. Our data thus show the discovery and validation of three new B. tabaci MEAM1 effectors that increase whitefly fecundity and modulate plant immunity. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.
Assuntos
Hemípteros , Nicotiana , Animais , Nicotiana/genética , Nicotiana/microbiologia , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/microbiologia , Solanum lycopersicum/parasitologia , Pseudomonas syringae/fisiologia , Doenças das Plantas/parasitologia , Doenças das Plantas/microbiologia , Reguladores de Crescimento de Plantas/metabolismo , Fertilidade/genéticaRESUMO
The sesquiterpenoid polygodial, which belongs to the drimane family, has been shown to be an antifeedant for a number of herbivorous insects. It is presumed to be synthesized from farnesyl diphosphate via drimenol, subsequent C-12 hydroxylation and further oxidations at both C-11 and C-12 to form a dialdehyde. Here, we have identified a drimenol synthase (PhDS) and a cytochrome P450 drimenol oxidase (PhDOX1) from Persicaria hydropiper. Expression of PhDS in yeast and plants resulted in production of drimenol alone. Co-expression of PhDS with PhDOX1 in yeast yielded drimendiol, the 12-hydroxylation product of drimenol, as a major product, and cinnamolide. When PhDS and PhDOX1 were transiently expressed by agro-infiltration in Nicotiana benthamiana leaves, drimenol was almost completely converted into cinnamolide and several additional drimenol derivatives were observed. In vitro assays showed that PhDOX1 only catalyses the conversion from drimenol to drimendiol, and not the further oxidation into an aldehyde. In yeast and heterologous plant hosts, the C-12 position of drimendiol is therefore likely to be further oxidized by endogenous enzymes into an aldehyde and subsequently converted to cinnamolide, presumably by spontaneous hemiacetal formation with the C-11 hydroxyl group followed by oxidation. Purified cinnamolide was confirmed by NMR and shown to be deterrent with an effective deterrent dose (ED50 ) of about 200-400 µg g-1 fresh weight against both whiteflies and aphids. The putative additional physiological and biochemical requirements for polygodial biosynthesis and stable storage in plant tissues are discussed.
Assuntos
Polygonaceae/enzimologia , Polygonaceae/metabolismo , Sesquiterpenos/metabolismo , Animais , Afídeos/efeitos dos fármacos , Hemípteros/efeitos dos fármacos , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , Folhas de Planta/enzimologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Sesquiterpenos Policíclicos , Polygonaceae/genética , Sesquiterpenos/farmacologia , Terpenos/metabolismo , Nicotiana/enzimologia , Nicotiana/genética , Nicotiana/metabolismoRESUMO
Plants have developed diverse defence mechanisms to ward off herbivorous pests. However, agriculture still faces estimated crop yield losses ranging from 25% to 40% annually. These losses arise not only because of direct feeding damage, but also because many pests serve as vectors of plant viruses. Herbivorous thrips (Thysanoptera) are important pests of vegetable and ornamental crops worldwide, and encompass virtually all general problems of pests: they are highly polyphagous, hard to control because of their complex lifestyle, and they are vectors of destructive viruses. Currently, control management of thrips mainly relies on the use of chemical pesticides. However, thrips rapidly develop resistance to these pesticides. With the rising demand for more sustainable, safer, and healthier food production systems, we urgently need to pinpoint the gaps in knowledge of plant defences against thrips to enable the future development of novel control methods. In this review, we summarize the current, rather scarce, knowledge of thrips-induced plant responses and the role of phytohormonal signalling and chemical defences in these responses. We describe concrete opportunities for breeding resistance against pests such as thrips as a prototype approach for next-generation resistance breeding.
Assuntos
Produtos Agrícolas/imunologia , Produtos Agrícolas/parasitologia , Doenças das Plantas/parasitologia , Proteínas de Plantas/imunologia , Tisanópteros/fisiologia , Animais , Produtos Agrícolas/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/genética , Transdução de SinaisRESUMO
In nature, plants have to cope with a wide range of stress conditions that often occur simultaneously or in sequence. To investigate how plants cope with multi-stress conditions, we analyzed the dynamics of whole-transcriptome profiles of Arabidopsis thaliana exposed to six sequential double stresses inflicted by combinations of: (i) infection by the necrotrophic fungus Botrytis cinerea, (ii) herbivory by chewing larvae of Pieris rapae, and (iii) drought stress. Each of these stresses induced specific expression profiles over time, in which one-third of all differentially expressed genes was shared by at least two single stresses. Of these, 394 genes were differentially expressed during all three stress conditions, albeit often in opposite directions. When two stresses were applied in sequence, plants displayed transcriptome profiles that were very similar to the second stress, irrespective of the nature of the first stress. Nevertheless, significant first-stress signatures could be identified in the sequential stress profiles. Bioinformatic analysis of the dynamics of co-expressed gene clusters highlighted specific clusters and biological processes of which the timing of activation or repression was altered by a prior stress. The first-stress signatures in second stress transcriptional profiles were remarkably often related to responses to phytohormones, strengthening the notion that hormones are global modulators of interactions between different types of stress. Because prior stresses can affect the level of tolerance against a subsequent stress (e.g. prior herbivory strongly affected resistance to B. cinerea), the first-stress signatures can provide important leads for the identification of molecular players that are decisive in the interactions between stress response pathways.
Assuntos
Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Estresse Fisiológico , Transcriptoma , Adaptação Biológica , Arabidopsis/metabolismo , Arabidopsis/fisiologia , RNA Mensageiro/metabolismo , RNA de Plantas/metabolismo , Análise de Sequência de RNARESUMO
In nature, plants are exposed to biotic and abiotic stresses that often occur simultaneously. Therefore, plant responses to combinations of stresses are most representative of how plants respond to stresses. We used RNAseq to assess temporal changes in the transcriptome of Arabidopsis thaliana to herbivory by Pieris rapae caterpillars, either alone or in combination with prior exposure to drought or infection with the necrotrophic fungus Botrytis cinerea. Pre-exposure to drought stress or Botrytis infection resulted in a significantly different timing of the caterpillar-induced transcriptional changes. Additionally, the combination of drought and P. rapae induced an extensive downregulation of A. thaliana genes involved in defence against pathogens. Despite a more substantial growth reduction observed for plants exposed to drought plus P. rapae feeding compared with P. rapae feeding alone, this did not affect weight increase of this specialist caterpillar. Plants respond to combined stresses with phenotypic and transcriptional changes that differ from the single stress situation. The effect of a previous exposure to drought or B. cinerea infection on transcriptional changes to caterpillars is largely overridden by the stress imposed by caterpillars, indicating that plants shift their response to the most recent stress applied.
Assuntos
Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/microbiologia , Transcriptoma , Animais , Arabidopsis/fisiologia , Botrytis/fisiologia , Borboletas/fisiologia , Secas , Herbivoria , Estresse FisiológicoRESUMO
There are few high-quality instruments to evaluate the participation and social functioning of fibromyalgia patients. The Fibromyalgia Participation Questionnaire (FPQ) is a questionnaire that evaluates these aspects with high reliability and validity in its German original version. The aim of this work was to describe the translation and cross-cultural adaptation process of the FPQ into Spanish and its validation to ensure the equivalence against the original version. The questionnaire will be translated according to the FACIT methodology, and it will be tested in the Clinical Management Unit of North Almeria Health Area. This methodology includes several stages: double forward translation, reconciled version, back-translation, review of the previous versions and development of the prefinal version for the pretest. Once the pretest ends, the final version of the questionnaire will be developed, which will be subjected to a validation process to study its psychometric properties. Reliability will be studied by internal consistency and test-retest reliability through Cronbach's alpha and Pearson's correlation coefficient, respectively. External and construct validity will be analysed using correlation coefficients, content validity with an empirical analysis, and a differential item functioning analysis will be employed to measure discriminative validity. The presence of ceiling and floor effects will be calculated too. The validation of the FPQ into different languages will allow better evaluation and treatment based on the observed limitations fibromyalgia patients suffer from, as well as bringing the possibility to compare between other countries and generalize its use in the scientific community.
Assuntos
Fibromialgia/psicologia , Projetos de Pesquisa , Inquéritos e Questionários , Protocolos Clínicos , Comparação Transcultural , Fibromialgia/diagnóstico , Humanos , Psicometria , TraduçõesRESUMO
Tomato breeding has been tremendously efficient in increasing fruit quality and quantity but did not focus on improving herbivore resistance. The biosynthetic pathway for the production of 7-epizingiberene in a wild tomato was introduced into a cultivated greenhouse variety with the aim to obtain herbivore resistance. 7-Epizingiberene is a specific sesquiterpene with toxic and repellent properties that is produced and stored in glandular trichomes. We identified 7-epizingiberene synthase (ShZIS) that belongs to a new class of sesquiterpene synthases, exclusively using Z-Z-farnesyl-diphosphate (zFPP) in plastids, probably arisen through neo-functionalization of a common ancestor. Expression of the ShZIS and zFPP synthases in the glandular trichomes of cultivated tomato resulted in the production of 7-epizingiberene. These tomatoes gained resistance to several herbivores that are pests of tomato. Hence, introduction of this sesquiterpene biosynthetic pathway into cultivated tomatoes resulted in improved herbivore resistance.
Assuntos
Vias Biossintéticas/genética , Herbivoria/imunologia , Engenharia Metabólica/métodos , Sesquiterpenos/imunologia , Solanum lycopersicum/imunologia , Alquil e Aril Transferases/genética , Alquil e Aril Transferases/metabolismo , Análise de Variância , Animais , Cruzamento/métodos , Clonagem Molecular , Comportamento Alimentar/fisiologia , Cromatografia Gasosa-Espectrometria de Massas , Hemípteros/fisiologia , Solanum lycopersicum/genética , Solanum lycopersicum/parasitologia , Sesquiterpenos Monocíclicos , Fosfatos de Poli-Isoprenil , Reação em Cadeia da Polimerase em Tempo Real , Sesquiterpenos/metabolismo , Tetranychidae/fisiologiaRESUMO
The complex interactions between aphids and their host plant are species-specific and involve multiple layers of recognition and defense. Aphid salivary proteins, which are released into the plant during phloem feeding, are a likely mediator of these interactions. In an approach to identify aphid effectors that facilitate feeding from host plants, eleven Myzus persicae (green peach aphid) salivary proteins and the GroEL protein of Buchnera aphidicola, a bacterial endosymbiont of this aphid species, were expressed transiently in Nicotiana tabacum (tobacco). Whereas two salivary proteins increased aphid reproduction, expression of three other aphid proteins and GroEL significantly decreased aphid reproduction on N. tabacum. These effects were recapitulated in stable transgenic Arabidopsis thaliana plants. Further experiments with A. thaliana expressing Mp55, a salivary protein that increased aphid reproduction, showed lower accumulation of 4-methoxyindol-3-ylmethylglucosinolate, callose and hydrogen peroxide in response to aphid feeding. Mp55-expressing plants also were more attractive for aphids in choice assays. Silencing Mp55 gene expression in M. persicae using RNA interference approaches reduced aphid reproduction on N. tabacum, A. thaliana, and N. benthamiana. Together, these results demonstrate a role for Mp55, a protein with as-yet-unknown molecular function, in the interaction of M. persicae with its host plants.
Assuntos
Afídeos/metabolismo , Proteínas de Insetos/metabolismo , Proteínas de Insetos/farmacologia , Nicotiana/metabolismo , Animais , Arabidopsis/fisiologia , Buchnera/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Folhas de Planta/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Reprodução , Nicotiana/genéticaRESUMO
Since research on plant interactions with herbivores and pathogens is often constrained by the analysis of already known compounds, there is a need to identify new defense-related plant metabolites. The uncommon nonprotein amino acid N(δ)-acetylornithine was discovered in a targeted search for Arabidopsis thaliana metabolites that are strongly induced by the phytohormone methyl jasmonate (MeJA). Stable isotope labeling experiments show that, after MeJA elicitation, Arg, Pro, and Glu are converted to Orn, which is acetylated by NATA1 to produce N(δ)-acetylornithine. MeJA-induced N(δ)-acetylornithine accumulation occurs in all tested Arabidopsis accessions, other Arabidopsis species, Capsella rubella, and Boechera stricta, but not in less closely related Brassicaceae. Both insect feeding and Pseudomonas syringae infection increase NATA1 expression and N(δ)-acetylornithine accumulation. NATA1 transient expression in Nicotiana tabacum and the addition of N(δ)-acetylornithine to an artificial diet both decrease Myzus persicae (green peach aphid) reproduction, suggesting a direct toxic or deterrent effect. However, since broad metabolic changes that are induced by MeJA in wild-type Arabidopsis are attenuated in a nata1 mutant strain, there may also be indirect effects on herbivores and pathogens. In the case of P. syringae, growth on a nata1 mutant is reduced compared with wild-type Arabidopsis, but growth in vitro is unaffected by N(δ)-acetylornithine addition.
Assuntos
Acetatos/farmacologia , Arabidopsis/metabolismo , Ciclopentanos/farmacologia , Ornitina/análogos & derivados , Oxilipinas/farmacologia , Acetilação , Animais , Afídeos/fisiologia , Arabidopsis/genética , Arabidopsis/microbiologia , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Herbivoria , Metaboloma , Dados de Sequência Molecular , Mutagênese Insercional , Mutação , Ornitina/biossíntese , Doenças das Plantas , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Pseudomonas syringae/patogenicidadeRESUMO
Inducible defenses, which provide enhanced resistance after initial attack, are nearly universal in plants. This defense signaling cascade is mediated by the synthesis, movement, and perception of jasmonic acid and related plant metabolites. To characterize the long-term persistence of plant immunity, we challenged Arabidopsis (Arabidopsis thaliana) and tomato (Solanum lycopersicum) with caterpillar herbivory, application of methyl jasmonate, or mechanical damage during vegetative growth and assessed plant resistance in subsequent generations. Here, we show that induced resistance was associated with transgenerational priming of jasmonic acid-dependent defense responses in both species, caused caterpillars to grow up to 50% smaller than on control plants, and persisted for two generations in Arabidopsis. Arabidopsis mutants that are deficient in jasmonate perception (coronatine insensitive1) or in the biogenesis of small interfering RNA (dicer-like2 dicer-like3 dicer-like4 and nuclear RNA polymerase d2a nuclear RNA polymerase d2b) do not exhibit inherited resistance. The observation of inherited resistance in both the Brassicaceae and Solanaceae suggests that this trait may be more widely distributed in plants. Epigenetic resistance to herbivory thus represents a phenotypically plastic mechanism for enhanced defense across generations.
Assuntos
Comportamento Alimentar , Insetos/fisiologia , Plantas/parasitologia , Acetatos/metabolismo , Animais , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Plantas/genética , RNA Interferente Pequeno/genéticaRESUMO
In most aphid species, facultative parthenogenetic reproduction allows rapid growth and formation of large single-genotype colonies. Upon predator attack, individual aphids emit an alarm pheromone to warn the colony of this danger. (E)-beta-farnesene (EBF) is the predominant constituent of the alarm pheromone in Myzus persicae (green peach aphid) and many other aphid species. Continuous exposure to alarm pheromone in aphid colonies raised on transgenic Arabidopsis thaliana plants that produce EBF leads to habituation within three generations. Whereas naive aphids are repelled by EBF, habituated aphids show no avoidance response. Similarly, individual aphids from the habituated colony can revert back to being EBF-sensitive in three generations, indicating that this behavioral change is not caused by a genetic mutation. Instead, DNA microarray experiments comparing gene expression in naive and habituated aphids treated with EBF demonstrate an almost complete desensitization in the transcriptional response to EBF. Furthermore, EBF-habituated aphids show increased progeny production relative to EBF-responsive aphids, with or without EBF treatment. Although both naive and habituated aphids emit EBF upon damage, EBF-responsive aphids have a higher survival rate in the presence of a coccinellid predator (Hippodamia convergens), and thus outperform habituated aphids that do not show an avoidance response. These results provide evidence that aphid perception of conspecific alarm pheromone aids in predator avoidance and thereby bestows fitness benefits in survivorship and fecundity. Therefore, although habituated M. persicae produce more progeny, EBF-emitting transgenic plants may have practical applications in agriculture as a result of increased predation of habituated aphids.
Assuntos
Afídeos/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Feromônios/farmacologia , Sesquiterpenos/farmacologia , Animais , Afídeos/fisiologia , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/parasitologia , Comportamento Animal/fisiologia , Cromatografia Gasosa-Espectrometria de Massas , Perfilação da Expressão Gênica , Habituação Psicofisiológica/fisiologia , Interações Hospedeiro-Parasita , Estrutura Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Feromônios/química , Feromônios/metabolismo , Plantas Geneticamente Modificadas , Sesquiterpenos/química , Sesquiterpenos/metabolismoRESUMO
Glucosinolates are biologically active natural products characteristic of crucifers, including oilseed rape, cabbage vegetables and the model plant Arabidopsis thaliana. Crucifer-specialist insect herbivores, like the economically important pest Plutella xylostella (diamondback moth), frequently use glucosinolates as oviposition stimuli. This suggests that the transfer of a glucosinolate biosynthetic pathway to a non-crucifer would stimulate oviposition on an otherwise non-attractive plant. Here, we demonstrate that stable genetic transfer of the six-step benzylglucosinolate pathway from A. thaliana to Nicotiana tabacum (tobacco) results in the production of benzylglucosinolate without causing morphological alterations. Benzylglucosinolate-producing tobacco plants were more attractive for oviposition by female P. xylostella moths than wild-type tobacco plants. As newly hatched P. xylostella larvae were unable to survive on tobacco, these results represent a proof-of-concept strategy for rendering non-host plants attractive for oviposition by specialist herbivores with the long-term goal of generating efficient dead-end trap crops for agriculturally important pests.
Assuntos
Produtos Agrícolas/genética , Engenharia Genética/métodos , Mariposas/fisiologia , Nicotiana/genética , Controle Biológico de Vetores , Feromônios/genética , Tiocianatos/metabolismo , Tioglucosídeos/metabolismo , Animais , Bioensaio , Vias Biossintéticas , Sistema Enzimático do Citocromo P-450/metabolismo , Feminino , Larva/crescimento & desenvolvimento , Mariposas/crescimento & desenvolvimento , Fases de Leitura Aberta/genética , Oviposição , Plantas Geneticamente Modificadas , Análise de Sobrevida , Nicotiana/crescimento & desenvolvimento , Nicotiana/parasitologia , Transformação GenéticaRESUMO
Glucosinolates are a diverse group of defensive secondary metabolites that is characteristic of the Brassicales. Arabidopsis thaliana (L.) Heynh. (Brassicaceae) lines with mutations that greatly reduce abundance of indole glucosinolates (cyp79B2 cyp79B3), aliphatic glucosinolates (myb28 myb29), or both (cyp79B2 cyp79B3 myb28 myb29) make it possible to test the in vivo defensive function of these two major glucosinolate classes. In experiments with Lepidoptera that are not crucifer-feeding specialists, aliphatic and indole glucosinolates had an additive effect on Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) larval growth, whereas Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) and Manduca sexta (L.) (Lepidoptera: Sphingidae) were affected only by the absence of aliphatic glucosinolates. In the case of two crucifer-feeding specialists, Pieris rapae (L.) (Lepidoptera: Pieridae) and Plutella xylostella (L.) (Lepidoptera: Plutellidae), there were no major changes in larval performance due to decreased aliphatic and/or indole glucosinolate content. Nevertheless, choice tests show that aliphatic and indole glucosinolates act in an additive manner to promote larval feeding of both species and P. rapae oviposition. Together, these results support the hypothesis that a diversity of glucosinolates is required to limit the growth of multiple insect herbivores.
Assuntos
Arabidopsis/química , Glucosinolatos/química , Glucosinolatos/farmacologia , Indóis/farmacologia , Lepidópteros/efeitos dos fármacos , Lepidópteros/fisiologia , Animais , Arabidopsis/genética , Arabidopsis/metabolismo , Comportamento Animal/efeitos dos fármacos , Dieta , Comportamento Alimentar , Feminino , Glucosinolatos/metabolismo , Lepidópteros/crescimento & desenvolvimento , MutaçãoRESUMO
Myzus persicae (green peach aphid) feeding on Arabidopsis thaliana induces a defence response, quantified as reduced aphid progeny production, in infested leaves but not in other parts of the plant. Similarly, infiltration of aphid saliva into Arabidopsis leaves causes only a local increase in aphid resistance. Further characterization of the defence-eliciting salivary components indicates that Arabidopsis recognizes a proteinaceous elicitor with a size between 3 and 10 kD. Genetic analysis using well-characterized Arabidopsis mutants shows that saliva-induced resistance against M. persicae is independent of the known defence signalling pathways involving salicylic acid, jasmonate and ethylene. Among 78 Arabidopsis genes that were induced by aphid saliva infiltration, 52 had been identified previously as aphid-induced, but few are responsive to the well-known plant defence signalling molecules salicylic acid and jasmonate. Quantitative PCR analyses confirm expression of saliva-induced genes. In particular, expression of a set of O-methyltransferases, which may be involved in the synthesis of aphid-repellent glucosinolates, was significantly up-regulated by both M. persicae feeding and treatment with aphid saliva. However, this did not correlate with increased production of 4-methoxyindol-3-ylmethylglucosinolate, suggesting that aphid salivary components trigger an Arabidopsis defence response that is independent of this aphid-deterrent glucosinolate.
Assuntos
Afídeos/fisiologia , Arabidopsis/metabolismo , Glucosinolatos/biossíntese , Saliva/química , Animais , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclopentanos/metabolismo , Etilenos/biossíntese , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Indóis , Metiltransferases/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Oxilipinas/metabolismo , RNA de Plantas/genética , Ácido Salicílico/metabolismo , Transdução de SinaisRESUMO
The diamondback moth (Plutella xylostella), a crucifer-specialist pest, has been documented to employ glucosinolates as host recognition cues for oviposition. Through the use of mutant Arabidopsis thaliana plants, we investigated the role of specific classes of glucosinolates in the signaling of oviposition by P. xylostella in vivo. Indole glucosinolate production in A. thaliana was found to be crucial in attracting oviposition. Additionally, indole glucosinolates functioned as oviposition cues only when in their intact form. 4-Methoxy-indol-3-ylmethylglucosinolate was implicated as an especially strong oviposition attractant in vitro, suggesting that indole glucosinolate secondary structure may play a role in P. xylostella host recognition as well. Aliphatic glucosinolate-derived breakdown products were found to attract P. xylostella, but only after damage or in the absence of indole glucosinolates. Furthermore, mutant plants lacking both intact indole glucosinolates and aliphatic glucosinolate breakdown products exhibited decreased oviposition attractiveness beyond that of the progenitor mutants lacking either component of the glucosinolate-myrosinase system. Therefore, we conclude that nonvolatile indole glucosinolates and volatile aliphatic glucosinolate breakdown products both appear to play important roles as host recognition cues for P. xylostella oviposition.
Assuntos
Arabidopsis/parasitologia , Glucosinolatos/metabolismo , Indóis/metabolismo , Mariposas/anatomia & histologia , Mariposas/fisiologia , Oviposição , Animais , Arabidopsis/genética , Arabidopsis/metabolismo , Sinais (Psicologia) , Glucosinolatos/química , Glucosinolatos/isolamento & purificação , Interações Hospedeiro-Parasita , Indóis/química , Indóis/isolamento & purificação , MutaçãoRESUMO
Whiteflies are among the world's most significant agricultural pests and chemical insecticides are extensively used to reduce crop damage to acceptable levels. However, nearly all insecticides pose a threat to the environment and alternative control methods, such as breeding of crop varieties that are inherently insect-resistant, are needed. Previously, a strong source of plant-age dependent resistance to the cabbage whitefly (Aleyrodes proletella) has been identified in the modern white cabbage (Brassica oleracea var. capitata) variety Rivera. However, nothing is known about the molecular mechanisms or the genes involved in this resistance. In the present study, a multidisciplinary approach combining transcriptome and metabolome profiling with genetic mapping was used to identify the molecular players of whitefly resistance in cabbage. Transcriptome profiles of young (susceptible) and older (resistant) Rivera plants were analyzed using RNA sequencing. While many genes involved in general processes were differentially expressed between both ages, several defense-related processes were overrepresented in the transcriptome profile of older plants. Hormone measurements revealed that jasmonic acid (JA) levels decreased upon whitefly infestation at both plant ages. Interestingly, abscisic acid (ABA) levels showed contrasting effects in response to whitefly infestation: ABA levels were reduced in young plants but induced in older plants upon whitefly feeding. Auxin levels were significantly lower in older plants compared with young plants, independent of whitefly presence, while glucosinolate levels were higher. Additionally, whitefly performance was monitored in an F2 population derived from a cross between Rivera and the susceptible white cabbage variety Christmas Drumhead. Significant QTL intervals were mapped on chromosome 2 and 9 for oviposition rate and whitefly adult survival, respectively. Several genes that were higher expressed in older plants and located in the identified QTL intervals were orthologous to Arabidopsis genes that have been related to ABA signaling, suggesting a role for ABA in the regulation of resistance towards whiteflies. Our results show that combining different omics approaches is a useful strategy to identify candidate genes underlying insect resistance.
Assuntos
Ácido Abscísico/metabolismo , Brassica/parasitologia , Mapeamento Cromossômico/métodos , Resistência à Doença , Hemípteros/fisiologia , Metaboloma/genética , Doenças das Plantas/parasitologia , Locos de Características Quantitativas/genética , Transcriptoma/genética , Animais , Brassica/genética , Brassica/crescimento & desenvolvimento , Cruzamentos Genéticos , Regulação da Expressão Gênica de Plantas , Glucosinolatos/metabolismo , Anotação de Sequência Molecular , Filogenia , Doenças das Plantas/genética , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/parasitologia , Análise de Componente Principal , Transdução de SinaisRESUMO
BACKGROUND: The green peach aphid, Myzus persicae (Sulzer), is a world-wide insect pest capable of infesting more than 40 plant families, including many crop species. However, despite the significant damage inflicted by M. persicae in agricultural systems through direct feeding damage and by its ability to transmit plant viruses, limited genomic information is available for this species. RESULTS: Sequencing of 16 M. persicae cDNA libraries generated 26,669 expressed sequence tags (ESTs). Aphids for library construction were raised on Arabidopsis thaliana, Nicotiana benthamiana, Brassica oleracea, B. napus, and Physalis floridana (with and without Potato leafroll virus infection). The M. persicae cDNA libraries include ones made from sexual and asexual whole aphids, guts, heads, and salivary glands. In silico comparison of cDNA libraries identified aphid genes with tissue-specific expression patterns, and gene expression that is induced by feeding on Nicotiana benthamiana. Furthermore, 2423 genes that are novel to science and potentially aphid-specific were identified. Comparison of cDNA data from three aphid lineages identified single nucleotide polymorphisms that can be used as genetic markers and, in some cases, may represent functional differences in the protein products. In particular, non-conservative amino acid substitutions in a highly expressed gut protease may be of adaptive significance for M. persicae feeding on different host plants. The Agilent eArray platform was used to design an M. persicae oligonucleotide microarray representing over 10,000 unique genes. CONCLUSION: New genomic resources have been developed for M. persicae, an agriculturally important insect pest. These include previously unknown sequence data, a collection of expressed genes, molecular markers, and a DNA microarray that can be used to study aphid gene expression. These resources will help elucidate the adaptations that allow M. persicae to develop compatible interactions with its host plants, complementing ongoing work illuminating plant molecular responses to phloem-feeding insects.
Assuntos
Afídeos/genética , Etiquetas de Sequências Expressas , Genoma de Inseto , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Polimorfismo de Nucleotídeo Único , Animais , Afídeos/crescimento & desenvolvimento , Catepsina B/genética , Cromossomos/genética , Biologia Computacional , Simulação por Computador , Bases de Dados de Ácidos Nucleicos , Perfilação da Expressão Gênica , Biblioteca Gênica , Genes de Insetos/genética , Genes Virais , Estágios do Ciclo de Vida , Repetições de Microssatélites/genética , Fenótipo , Vírus de Plantas/genética , Reprodutibilidade dos Testes , Proteínas e Peptídeos Salivares/genética , Proteínas e Peptídeos Salivares/metabolismo , Análise de Sequência de DNARESUMO
Plant defenses against pathogens and insects are regulated differentially by cross-communicating signaling pathways in which salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) play key roles. To understand how plants integrate pathogen- and insect-induced signals into specific defense responses, we monitored the dynamics of SA, JA, and ET signaling in Arabidopsis after attack by a set of microbial pathogens and herbivorous insects with different modes of attack. Arabidopsis plants were exposed to a pathogenic leaf bacterium (Pseudomonas syringae pv. tomato), a pathogenic leaf fungus (Alternaria brassicicola), tissue-chewing caterpillars (Pieris rapae), cell-content-feeding thrips (Frankliniella occidentalis), or phloem-feeding aphids (Myzus persicae). Monitoring the signal signature in each plant-attacker combination showed that the kinetics of SA, JA, and ET production varies greatly in both quantity and timing. Analysis of global gene expression profiles demonstrated that the signal signature characteristic of each Arabidopsis-attacker combination is orchestrated into a surprisingly complex set of transcriptional alterations in which, in all cases, stress-related genes are overrepresented. Comparison of the transcript profiles revealed that consistent changes induced by pathogens and insects with very different modes of attack can show considerable overlap. Of all consistent changes induced by A. brassicicola, Pieris rapae, and E occidentalis, more than 50% also were induced consistently by P. syringae. Notably, although these four attackers all stimulated JA biosynthesis, the majority of the changes in JA-responsive gene expression were attacker specific. All together, our study shows that SA, JA, and ET play a primary role in the orchestration of the plant's defense response, but other regulatory mechanisms, such as pathway cross-talk or additional attacker-induced signals, eventually shape the highly complex attacker-specific defense response.
Assuntos
Arabidopsis/genética , Arabidopsis/fisiologia , Doenças das Plantas/genética , Alternaria/patogenicidade , Animais , Arabidopsis/microbiologia , Arabidopsis/parasitologia , Sequência de Bases , Ciclopentanos/metabolismo , DNA de Plantas/genética , Etilenos/metabolismo , Perfilação da Expressão Gênica , Genes de Plantas , Marcadores Genéticos , Insetos/patogenicidade , Análise de Sequência com Séries de Oligonucleotídeos , Oxilipinas , Doenças das Plantas/microbiologia , Doenças das Plantas/parasitologia , Plantas Geneticamente Modificadas , Pseudomonas syringae/patogenicidade , Ácido Salicílico/metabolismo , Transdução de Sinais , Transcrição GênicaRESUMO
Plants and herbivores have co-evolved in their natural habitats for about 350 million years, but since the domestication of crops, plant resistance against insects has taken a different turn. With the onset of monoculture-driven modern agriculture, selective pressure on insects to overcome resistances has dramatically increased. Therefore plant breeders have resorted to high-tech tools to continuously create new insect-resistant crops. Efforts in the past 30 years have resulted in elucidation of mechanisms of many effective plant defenses against insect herbivores. Here, we critically appraise these efforts and - with a focus on sap-sucking insects - discuss how these findings have contributed to herbivore-resistant crops. Moreover, in this review we try to assess where future challenges and opportunities lay ahead. Of particular importance will be a mandatory reduction in systemic pesticide usage and thus a greater reliance on alternative methods, such as improved plant genetics for plant resistance to insect herbivores.
RESUMO
Plants in their natural environments are constantly subjected to biotic stress. In addition to possessing physical barriers and anti-nutritive toxins, plants can be primed to respond more efficiently against future attack via faster and stronger gene activation. Here we discuss recent findings showing that plants can pass signatures of attack to the next generation, thus rendering the progeny more resistant against insect and pathogen attack. A combination of phytohormone signaling, small RNA-mediated gene silencing and DNA methylation are involved in transgenerational induced resistance. Epiallelic variation against biotic threats should be under positive selection in populations of plants where the environment is predictable over time. Similarly, in very genetically homogenous populations, such as during range expansion, epigenome reorganization is a likely mechanism for faster plant adaptation to novel biotic attack. Further research is needed to understand the relative role of the genome vs. the epigenome for the evolution of increased plant resistance.