RESUMO
Type-IV glandular trichomes, which only occur in the juvenile developmental phase of the cultivated tomato (Solanum lycopersicum), produce acylsugars that broadly protect against arthropod herbivory. Previously, we introgressed the capacity to retain type-IV trichomes in the adult phase from the wild tomato, Solanum galapagense, into the cultivated species cv. Micro-Tom (MT). The resulting MT-Galapagos enhanced trichome (MT-Get) introgression line contained 5 loci associated with enhancing the density of type-IV trichomes in adult plants. We genetically dissected MT-Get and obtained a subline containing only the locus on Chromosome 2 (MT-Get02). This genotype displayed about half the density of type-IV trichomes compared to the wild progenitor. However, when we stacked the gain-of-function allele of WOOLLY, which encodes a homeodomain leucine zipper IV transcription factor, Get02/Wo exhibited double the number of type-IV trichomes compared to S. galapagense. This discovery corroborates previous reports positioning WOOLLY as a master regulator of trichome development. Acylsugar levels in Get02/Wo were comparable to the wild progenitor, although the composition of acylsugar types differed, especially regarding fewer types with medium-length acyl chains. Agronomical parameters of Get02/Wo, including yield, were comparable to MT. Pest resistance assays showed enhanced protection against silverleaf whitefly (Bemisia tabaci), tobacco hornworm (Manduca sexta), and the fungus Septoria lycopersici. However, resistance levels did not reach those of the wild progenitor, suggesting the specificity of acylsugar types in the pest resistance mechanism. Our findings in trichome-mediated resistance advance the development of robust, naturally resistant tomato varieties, harnessing the potential of natural genetic variation. Moreover, by manipulating only 2 loci, we achieved exceptional results for a highly complex, polygenic trait, such as herbivory resistance in tomato.
Assuntos
Solanum lycopersicum , Tricomas , Tricomas/genética , Solanum lycopersicum/genética , Solanum lycopersicum/parasitologia , Animais , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Mutação/genética , Herbivoria , Herança Multifatorial , Manduca/fisiologia , Doenças das Plantas/genética , Doenças das Plantas/parasitologiaRESUMO
Secretions from glandular trichomes potentially protect plants against a variety of aggressors. In the tomato clade of the Solanum genus, glandular trichomes of wild species produce a rich source of chemical diversity at the leaf surface. Previously, 7-epi-zingiberene produced in several accessions of Solanum habrochaites was found to confer resistance to whiteflies (Bemisia tabaci) and other insect pests. Here, we report the identification and characterisation of 9-hydroxy-zingiberene (9HZ) and 9-hydroxy-10,11-epoxyzingiberene (9H10epoZ), two derivatives of 7-epi-zingiberene produced in glandular trichomes of S. habrochaites LA2167. Using a combination of transcriptomics and genetics, we identified a gene coding for a cytochrome P450 oxygenase, ShCYP71D184, that is highly expressed in trichomes and co-segregates with the presence of the zingiberene derivatives. Transient expression assays in Nicotiana benthamiana showed that ShCYP71D184 carries out two successive oxidations to generate 9HZ and 9H10epoZ. Bioactivity assays showed that 9-hydroxy-10,11-epoxyzingiberene in particular exhibits substantial toxicity against B. tabaci and various microorganisms including Phytophthora infestans and Botrytis cinerea. Our work shows that trichome secretions from wild tomato species can provide protection against a wide variety of organisms. In addition, the availability of the genes encoding the enzymes for the pathway of 7-epi-zingiberene derivatives makes it possible to introduce this trait in cultivated tomato by precision breeding.
Assuntos
Hemípteros/metabolismo , Sesquiterpenos Monocíclicos/metabolismo , NADPH-Ferri-Hemoproteína Redutase/metabolismo , Solanum/metabolismo , Animais , Botrytis/efeitos dos fármacos , Botrytis/patogenicidade , Hemípteros/genética , Hemípteros/microbiologia , Sesquiterpenos Monocíclicos/toxicidade , NADPH-Ferri-Hemoproteína Redutase/genética , Phytophthora infestans/efeitos dos fármacos , Phytophthora infestans/patogenicidade , Solanum/genéticaRESUMO
BACKGROUND: Plant-produced specialised metabolites are a powerful part of a plant's first line of defence against herbivorous insects, bacteria and fungi. Wild ancestors of present-day cultivated tomato produce a plethora of acylsugars in their type-I/IV trichomes and volatiles in their type-VI trichomes that have a potential role in plant resistance against insects. However, metabolic profiles are often complex mixtures making identification of the functionally interesting metabolites challenging. Here, we aimed to identify specialised metabolites from a wide range of wild tomato genotypes that could explain resistance to vector insects whitefly (Bemisia tabaci) and Western flower thrips (Frankliniella occidentalis). We evaluated plant resistance, determined trichome density and obtained metabolite profiles of the glandular trichomes by LC-MS (acylsugars) and GC-MS (volatiles). Using a customised Random Forest learning algorithm, we determined the contribution of specific specialised metabolites to the resistance phenotypes observed. RESULTS: The selected wild tomato accessions showed different levels of resistance to both whiteflies and thrips. Accessions resistant to one insect can be susceptible to another. Glandular trichome density is not necessarily a good predictor for plant resistance although the density of type-I/IV trichomes, related to the production of acylsugars, appears to correlate with whitefly resistance. For type VI-trichomes, however, it seems resistance is determined by the specific content of the glands. There is a strong qualitative and quantitative variation in the metabolite profiles between different accessions, even when they are from the same species. Out of 76 acylsugars found, the random forest algorithm linked two acylsugars (S3:15 and S3:21) to whitefly resistance, but none to thrips resistance. Out of 86 volatiles detected, the sesquiterpene α-humulene was linked to whitefly susceptible accessions instead. The algorithm did not link any specific metabolite to resistance against thrips, but monoterpenes α-phellandrene, α-terpinene and ß-phellandrene/D-limonene were significantly associated with susceptible tomato accessions. CONCLUSIONS: Whiteflies and thrips are distinctly targeted by certain specialised metabolites found in wild tomatoes. The machine learning approach presented helped to identify features with efficacy toward the insect species studied. These acylsugar metabolites can be targets for breeding efforts towards the selection of insect-resistant cultivars.
Assuntos
Resistência à Doença/genética , Variação Genética , Hemípteros/fisiologia , Metaboloma/genética , Solanum/genética , Tisanópteros/fisiologia , Tricomas/genética , Tricomas/metabolismo , Algoritmos , Animais , Ecótipo , Genótipo , Fenótipo , Compostos Orgânicos Voláteis/análiseRESUMO
MAIN CONCLUSION: Cultivated tomatoes harboring the plastid-derived sesquiterpenes from S. habrochaites have altered type-VI trichome morphology and unveil additional genetic components necessary for piercing-sucking pest resistance. Arthropod resistance in the tomato wild relative Solanum habrochaites LA1777 is linked to specific sesquiterpene biosynthesis. The Sesquiterpene synthase 2 (SsT2) gene cluster on LA1777 chromosome 8 controls plastid-derived sesquiterpene synthesis. The main genes at SsT2 are Z-prenyltransferase (zFPS) and Santalene and Bergamotene Synthase (SBS), which produce α-santalene, ß-bergamotene, and α-bergamotene in LA1777 round-shaped type-VI glandular trichomes. Cultivated tomatoes have mushroom-shaped type-VI trichomes with much smaller glands that contain low levels of monoterpenes and cytosolic-derived sesquiterpenes, not presenting the same pest resistance as in LA1777. We successfully transferred zFPS and SBS from LA1777 to cultivated tomato (cv. Micro-Tom, MT) by a backcrossing approach. The trichomes of the MT-Sst2 introgressed line produced high levels of the plastid-derived sesquiterpenes. The type-VI trichome internal storage-cavity size increased in MT-Sst2, probably as an effect of the increased amount of sesquiterpenes, although it was not enough to mimic the round-shaped LA1777 trichomes. The presence of high amounts of plastid-derived sesquiterpenes was also not sufficient to confer resistance to various tomato piercing-sucking pests, indicating that the effect of the sesquiterpenes found in the wild S. habrochaites can be insect specific. Our results provide for a better understanding of the morphology of S. habrochaites type-VI trichomes and paves the way to obtain insect-resistant tomatoes.
Assuntos
Artrópodes , Sesquiterpenos , Solanum lycopersicum , Solanum , Animais , Solanum lycopersicum/genética , Solanum/genética , TricomasRESUMO
In this paper, we describe why and how to build a local community of practice in scientific programming for life scientists who use computers and programming in their research. A community of practice is a small group of scientists who meet regularly to help each other and promote good practices in scientific programming. While most life scientists are well trained in the laboratory to conduct experiments, good practices with (big) data sets and their analysis are often missing. We propose a model on how to build such a community of practice at a local academic institution, present two real-life examples, and introduce challenges and implemented solutions. We believe that the current data deluge that life scientists face can benefit from the implementation of these small communities. Good practices spread among experimental scientists will foster open, transparent, and sound scientific results beneficial to society.
Assuntos
Participação da Comunidade/métodos , Ciência de Dados/métodos , Big Data , Análise de Dados , Educação Profissionalizante , Humanos , Modelos Teóricos , Pesquisa , Projetos de Pesquisa/normasRESUMO
Volatiles mediate the interaction of plants with pollinators, herbivores and their natural enemies, other plants and micro-organisms. With increasing knowledge about these interactions the underlying mechanisms turn out to be increasingly complex. The mechanisms of biosynthesis and perception of volatiles are slowly being uncovered. The increasing scientific knowledge can be used to design and apply volatile-based agricultural strategies.
Assuntos
Resistência à Doença/imunologia , Herbivoria , Plantas/metabolismo , Polinização , Compostos Orgânicos Voláteis/química , Agricultura , Ciclopentanos/química , Ciclopentanos/metabolismo , Resistência à Doença/fisiologia , Regulação da Expressão Gênica de Plantas/imunologia , Regulação da Expressão Gênica de Plantas/fisiologia , Oxilipinas/química , Oxilipinas/metabolismo , Fenóis/química , Fenóis/metabolismo , Plantas/química , Plantas/microbiologia , Plantas/parasitologia , Transdução de Sinais/fisiologia , Terpenos/química , Terpenos/metabolismo , Compostos Orgânicos Voláteis/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
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
Type-VI glandular trichomes of wild tomato Solanum habrochaites PI127826 produce high levels of the sesquiterpene 7-epizingiberene and its derivatives, making the plant repellent and toxic to several pest insects and pathogens. How wild tomato trichomes achieve such high terpene production is still largely unknown. Here we show that a cross (F1) with a cultivated tomato produced only minute levels of 7-epizingiberene. In the F2-progeny, selected for the presence of the 7-epizingiberene biosynthesis genes, only three percent produced comparable amounts the wild parent, indicating this trait is recessive and multigenic. Moreover, trichome density alone did not explain the total levels of terpene levels found on the leaves. We selected F2 plants with the "high-production active-trichome phenotype" of PI127826, having trichomes producing about 150 times higher levels of terpenes than F2 individuals that displayed a "low-production lazy-trichome phenotype". Terpene quantities in trichomes of these F2 plants correlated with the volume of the storage cavity and shape of the gland. We found that trichome morphology is not a predetermined characteristic, but cavity volume rather depended on gland-cell metabolic activity. Inhibitor assays showed that the plastidial-precursor pathway (MEP) is fundamental for high-level production of both cytosolic as well as plastid-derived terpenes in tomato trichomes. Additionally, gene expression profiles of isolated secretory cells showed that key enzymes in the MEP pathway were higher expressed in active trichomes. We conclude that the MEP pathway is the primary precursor-supply route in wild tomato type-VI trichomes and that the high-production phenotype of the wild tomato trichome is indeed a multigenic trait.
RESUMO
Compounds of the terpenoid class play numerous roles in the interactions of plants with their environment, such as attracting pollinators and defending the plant against pests. We show here that the genome of cultivated tomato (Solanum lycopersicum) contains 44 terpene synthase (TPS) genes, including 29 that are functional or potentially functional. Of these 29 TPS genes, 26 were expressed in at least some organs or tissues of the plant. The enzymatic functions of eight of the TPS proteins were previously reported, and here we report the specific in vitro catalytic activity of 10 additional tomato terpene synthases. Many of the tomato TPS genes are found in clusters, notably on chromosomes 1, 2, 6, 8, and 10. All TPS family clades previously identified in angiosperms are also present in tomato. The largest clade of functional TPS genes found in tomato, with 12 members, is the TPS-a clade, and it appears to encode only sesquiterpene synthases, one of which is localized to the mitochondria, while the rest are likely cytosolic. A few additional sesquiterpene synthases are encoded by TPS-b clade genes. Some of the tomato sesquiterpene synthases use z,z-farnesyl diphosphate in vitro as well, or more efficiently than, the e,e-farnesyl diphosphate substrate. Genes encoding monoterpene synthases are also prevalent, and they fall into three clades: TPS-b, TPS-g, and TPS-e/f. With the exception of two enzymes involved in the synthesis of ent-kaurene, the precursor of gibberellins, no other tomato TPS genes could be demonstrated to encode diterpene synthases so far.
Assuntos
Alquil e Aril Transferases/genética , Proteínas de Plantas/genética , Solanum lycopersicum/genética , Alquil e Aril Transferases/metabolismo , Ciclopentanos/farmacologia , Citosol/enzimologia , Diterpenos do Tipo Caurano/metabolismo , Evolução Molecular , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/enzimologia , Mitocôndrias/enzimologia , Dados de Sequência Molecular , Monoterpenos/metabolismo , Família Multigênica , Oxilipinas/farmacologia , Proteínas de Plantas/metabolismoRESUMO
Phytopathogens and herbivores induce plant defences. Whereas there is evidence that some pathogens suppress these defences by interfering with signalling pathways involved in the defence, such evidence is scarce for herbivores. We found that the invasive spider mite Tetranychus evansi suppresses the induction of the salicylic acid and jasmonic acid signalling routes involved in induced plant defences in tomato. This was reflected in the levels of inducible defence compounds, such as proteinase inhibitors, which in mite-infested plants were reduced to even lower levels than the constitutive levels in herbivore-free plants. Additionally, the spider mite suppressed the release of inducible volatiles, which are implicated in plant defence. Consequently, the mites performed much better on previously attacked plants than on non-attacked plants. These findings provide a new perspective on plant-herbivore interactions, plant protection and plant resistance to invasive species.
Assuntos
Cadeia Alimentar , Transdução de Sinais , Solanum lycopersicum/fisiologia , Tetranychidae/fisiologia , Animais , Ciclopentanos/análise , Ciclopentanos/metabolismo , Comportamento Alimentar , Feminino , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Espécies Introduzidas , Oxilipinas/análise , Oxilipinas/metabolismo , Doenças das Plantas/genética , Folhas de Planta/química , Dinâmica Populacional , Inibidores de Proteases/análise , Inibidores de Proteases/metabolismo , Distribuição Aleatória , Reprodução , Ácido Salicílico/análise , Ácido Salicílico/metabolismo , Tetranychidae/crescimento & desenvolvimentoRESUMO
Solanum lycopersicum and Solanum habrochaites (f. typicum) accession PI127826 emit a variety of sesquiterpenes. To identify terpene synthases involved in the production of these volatile sesquiterpenes, we used massive parallel pyrosequencing (RNA-seq) to obtain the transcriptome of the stem trichomes from these plants. This approach resulted initially in the discovery of six sesquiterpene synthase cDNAs from S. lycopersicum and five from S. habrochaites. Searches of other databases and the S. lycopersicum genome resulted in the discovery of two additional sesquiterpene synthases expressed in trichomes. The sesquiterpene synthases from S. lycopersicum and S. habrochaites have high levels of protein identity. Several of them appeared to encode for non-functional proteins. Functional recombinant proteins produced germacrenes, ß-caryophyllene/α-humulene, viridiflorene and valencene from (E,E)-farnesyl diphosphate. However, the activities of these enzymes do not completely explain the differences in sesquiterpene production between the two tomato plants. RT-qPCR confirmed high levels of expression of most of the S. lycopersicum sesquiterpene synthases in stem trichomes. In addition, one sesquiterpene synthase was induced by jasmonic acid, while another appeared to be slightly repressed by the treatment. Our data provide a foundation to study the evolution of terpene synthases in cultivated and wild tomato.
Assuntos
Alquil e Aril Transferases/química , Proteínas de Plantas/química , RNA de Plantas/química , Solanum lycopersicum/genética , Solanum/genética , Alquil e Aril Transferases/genética , DNA Complementar/química , Biblioteca Gênica , Solanum lycopersicum/enzimologia , Proteínas de Plantas/genética , Proteínas Recombinantes de Fusão/metabolismo , Análise de Sequência de RNA , Sesquiterpenos/metabolismo , Solanum/enzimologiaRESUMO
The phloem-feeding whitefly Bemisia tabaci is a serious pest to a broad range of host plants, including many economically important crops such as tomato. These insects serve as a vector for various devastating plant viruses. It is known that whiteflies are capable of manipulating host-defense responses, potentially mediated by effector molecules in the whitefly saliva. We hypothesized that, beside putative effector proteins, small RNAs (sRNA) are delivered by B. tabaci into the phloem, where they may play a role in manipulating host plant defenses. There is already evidence to suggest that sRNAs can mediate the host-pathogen dialogue. It has been shown that Botrytis cinerea, the causal agent of gray mold disease, takes advantage of the plant sRNA machinery to selectively silence host genes involved in defense signaling. Here we identified sRNAs originating from B. tabaci in the phloem of tomato plants on which they are feeding. sRNAs were isolated and sequenced from tomato phloem of whitefly-infested and control plants as well as from the nymphs themselves, control leaflets, and from the infested leaflets. Using stem-loop RT-PCR, three whitefly sRNAs have been verified to be present in whitefly-infested leaflets that were also present in the whitefly-infested phloem sample. Our results show that whitefly sRNAs are indeed present in tomato tissues upon feeding, and they appear to be mobile in the phloem. Their role in the host-insect interaction can now be investigated.
RESUMO
⢠Arsenate tolerance, uptake and arsenate-induced phytochelatin (PC) accumulation were compared at different phosphorus supply rates in two populations of the broom, Cytisus striatus , one from an arsenic-enriched gold mine and one from a nonmetalliferous site. ⢠After 7 d of exposure, arsenate tolerance was higher in the mine population. Arsenate uptake was phosphate-suppressible, and much lower in the mine plants. When compared at equal levels of stress, the mine plants and the nonmetallicolous plants exhibited similar arsenic accumulation, suggesting that reduced arsenate uptake is mainly responsible for superior tolerance. ⢠Arsenate-induced PC accumulation occurred in both plant types. The γ-glutamylcysteine synthetase inhibitor, L-buthioninesulfoximine, caused arsenate hypersensitivity in both plant types, suggesting that PC-based arsenic sequestration is essential for both normal and enhanced arsenate tolerance. Mine plants produced longer PCs than the nonmetallicolous plants, possibly due to a differential temporal pattern of arsenate accumulation. ⢠Our results are consistent with a similar mechanism underlying arsenate hypertolerance in C. striatus and grasses, that is reduced arsenate uptake through suppression of phosphate transporter activity.
RESUMO
A combination of metal immobilising agents and metal tolerant plants has been utilised in order to reduce the environmental impact of the acidic metal contaminated Jales mine spoil tips. The addition of Beringite (a modified aluminosilicate), steel shots (iron bearing material) and organic matter as spoil amendments resulted in changes in arsenate (As) concentrations and pH of spoil material and improved plant growth. The application of Beringite increased both pH and plant available As concentrations. A 4-year follow up of the spoil analysis demonstrated that the effect of the spoil treatments was stable following treatments, however, the effectiveness did not increase any more after 2 years. The use of metal tolerant grasses in combination with spoil treatments resulted in a rapid and effective revegetation of the As contaminated Jales mine spoils. Colonisation and reproduction of Agrostis castellana and Holcus lanatus was most successful when the substrate contained a combination of all three additives. Plant performances could be enhanced by supplementation of a phosphate fertiliser. The rapid reproduction of the two grass species makes them very suitable for revegetation purposes. Agrostis castellana and Holcus lanatus apparently exhibited a level of metal- and As-tolerance sufficient for survival on untreated spoil, but in the first stages of revegetation the use of spoil amendments was found to be essential. Organic matter in combination with Beringite and/or steel shots resulted in decreased As in the aboveground biomass, protecting possible grazers and predators from undesirable levels of As.
Assuntos
Arsênio/farmacocinética , Poaceae/crescimento & desenvolvimento , Poluentes do Solo/farmacocinética , Silicatos de Alumínio/química , Arsênio/isolamento & purificação , Biomassa , Conservação dos Recursos Naturais , Poluição Ambiental/prevenção & controle , Fertilizantes , Concentração de Íons de Hidrogênio , Mineração , Reprodução , Poluentes do Solo/isolamento & purificação , AçoRESUMO
How whiteflies (Bemisia tabaci) make the choice for a host plant prior to landing, is not precisely known. Here we investigated whether they respond to specific volatiles of tomato. Zingiberene and curcumene were purified from Solanum habrochaites (PI127826), characterised by NMR and X-ray analysis and identified as 7-epizingiberene and R-curcumene. In contrast, oil from Zingiber officinalis contained the stereoisomers zingiberene and S-curcumene, respectively. Using a combination of free-choice bio-assays and electroantennography, 7-epizingiberene and its dehydrogenated derivative R-curcumene were shown to be active as semiochemicals to B. tabaci adults, whereas the stereoisomers from ginger were not. In addition, R-curcumene elicited the strongest electroantennographic response. Bio-assays showed that a cultivated tomato could be made less attractive to B. tabaci than its neighbouring siblings by the addition of the tomato stereoisomer 7-epizingiberene or its derivative R-curcumene. These sesquiterpenes apparently repel adult whiteflies prior to landing, presumably because it informs them that after landing they, or their offspring, may be exposed to higher and lethal concentrations of the same compounds.
Assuntos
Hemípteros/fisiologia , Repelentes de Insetos/isolamento & purificação , Repelentes de Insetos/farmacologia , Sesquiterpenos/isolamento & purificação , Solanum lycopersicum/química , Animais , Hemípteros/efeitos dos fármacos , Interações Hospedeiro-Parasita , Repelentes de Insetos/química , Estrutura Molecular , Sesquiterpenos Monocíclicos , Óleos Voláteis , Sesquiterpenos/química , Sesquiterpenos/farmacologia , EstereoisomerismoRESUMO
Bemisia tabaci (whitefly) infestations and the subsequent transfer of viruses are the cause of severe losses in crop production and horticultural practice. To improve biological control of B. tabaci, we investigated repellent properties of plant-produced semiochemicals. The mix of headspace volatiles, collected from naturally repellent wild tomato accessions, influenced B. tabaci initial choice behavior, indicating a role for plant semiochemicals in locating host plants. A collection of wild tomato accessions and introgression lines (Solanum pennellii LA716 x Solanum lycopersicum 'Moneyberg') were extensively screened for attractiveness to B. tabaci, and their headspace profiles were determined by means of gas chromatography-mass spectrometry. Correlation analysis revealed that several terpenoids were putatively involved in tomato-whitefly interactions. Several of these candidate compounds conferred repellence to otherwise attractive tomato plants when applied to the plant's branches on paper cards. The sesquiterpenes zingiberene and curcumene and the monoterpenes p-cymene, alpha-terpinene, and alpha-phellandrene had the strongest effects in free-choice bioassays. These terpenes also elicited a response of receptors on the insect's antennae as determined by electroantennography. Conversely, the monoterpene beta-myrcene showed no activity in both assays. B. tabaci apparently uses, besides visual cues, specific plant volatile cues for the initial selection of a host. Altering whitefly choice behavior by manipulation of the terpenoid composition of the host headspace may therefore be feasible.
Assuntos
Hemípteros/fisiologia , Interações Hospedeiro-Parasita , Solanum lycopersicum/parasitologia , Compostos Orgânicos Voláteis/metabolismo , Animais , Comportamento Animal/classificação , Sinais (Psicologia) , Hemípteros/efeitos dos fármacos , Interações Hospedeiro-Parasita/efeitos dos fármacos , Endogamia , Repelentes de Insetos/farmacologia , Solanum lycopersicum/efeitos dos fármacos , Feromônios/farmacologia , Especificidade da EspécieRESUMO
Decreased arsenate [As(V)] uptake is the major mechanism of naturally selected As(V) hypertolerance in plants. However, As(V)-hypertolerant ecotypes also show enhanced rates of phytochelatin (PC) accumulation, suggesting that improved sequestration might additionally contribute to the hypertolerance phenotype. Here, we show that enhanced PC-based sequestration in As(V)-hypertolerant Holcus lanatus is not due to an enhanced capacity for PC synthesis as such, but to increased As(V) reductase activity. Vacuolar transport of arsenite-thiol complexes was equal in both ecotypes. Based on homology with the yeast As(V) reductase, Acr2p, we identified a Cdc25-like plant candidate, HlAsr, and confirmed the As(V) reductase activity of both HlAsr and the homologous protein from Arabidopsis thaliana. The gene appeared to be As(V)-inducible and its expression was enhanced in the As(V)-hypertolerant H. lanatus ecotype, compared with the non-tolerant ecotype. Homologous ectopic overexpression of the AtASR cDNA in A. thaliana produced a dual phenotype. It improved tolerance to mildly toxic levels of As(V) exposure, but caused hypersensitivity to more toxic levels. Arabidopsis asr T-DNA mutants showed increased As(V) sensitivity at low exposure levels and enhanced arsenic retention in the root. It is argued that, next to decreased uptake, enhanced expression of HlASR might act as an additional determinant of As(V) hypertolerance and As transport in H. lanatus.
Assuntos
Arseniatos/metabolismo , Glutationa/metabolismo , Holcus/enzimologia , Proteínas de Plantas/metabolismo , Fosfatases cdc25/metabolismo , Sequência de Aminoácidos , Análise de Variância , Arabidopsis/enzimologia , Arabidopsis/genética , Arseniatos/farmacologia , ATPases Transportadoras de Arsenito , Sequência Consenso , DNA Bacteriano/genética , DNA Complementar/metabolismo , Holcus/efeitos dos fármacos , Holcus/genética , Bombas de Íon/genética , Bombas de Íon/metabolismo , Dados de Sequência Molecular , Complexos Multienzimáticos/genética , Complexos Multienzimáticos/metabolismo , Mutagênese Insercional , Fenótipo , Fitoquelatinas , Proteínas de Plantas/genética , Alinhamento de Sequência , Fosfatases cdc25/genéticaRESUMO
Using the gamma-glutamylcysteine synthetase inhibitor, L-buthionine-[S,R]-sulphoximine (BSO), the role for phytochelatins (PCs) was evaluated in Cu, Cd, Zn, As, Ni, and Co tolerance in non-metallicolous and metallicolous, hypertolerant populations of Silene vulgaris (Moench) Garcke, Thlaspi caerulescens J.&C. Presl., Holcus lanatus L., and Agrostis castellana Boiss. et Reuter. Based on plant-internal PC-thiol to metal molar ratios, the metals' tendency to induce PC accumulation decreased in the order As/Cd/Cu > Zn > Ni/Co, and was consistently higher in non-metallicolous plants than in hypertolerant ones, except for the case of As. The sensitivities to Cu, Zn, Ni, and Co were consistently unaffected by BSO treatment, both in non-metallicolous and hypertolerant plants, suggesting that PC-based sequestration is not essential for constitutive tolerance or hypertolerance to these metals. Cd sensitivity was considerably increased by BSO, though exclusively in plants lacking Cd hypertolerance, suggesting that adaptive cadmium hypertolerance is not dependent on PC-mediated sequestration. BSO dramatically increased As sensitivity, both in non-adapted and As-hypertolerant plants, showing that PC-based sequestration is essential for both normal constitutive tolerance and adaptive hypertolerance to this metalloid. The primary function of PC synthase in plants and algae remains elusive.