RESUMO
The adoption of pest-resistant transgenic plants to reduce yield losses and decrease pesticide use has been successful. To achieve the goal of controlling both chewing and sucking pests in a given transgenic plant, we generated transgenic tobacco, Arabidopsis, and rice plants expressing the fusion protein, AaIT/GNA, in which an insecticidal scorpion venom neurotoxin (Androctonus australis toxin, AaIT) is fused to snowdrop lectin (Galanthus nivalis agglutinin, GNA). Compared with transgenic tobacco and Arabidopsis plants expressing AaIT or GNA, transgenic plants expressing AaIT/GNA exhibited increased resistance and toxicity to one chewing pest, the cotton bollworm, Helicoverpa armigera. Transgenic tobacco and rice plants expressing AaIT/GNA showed increased resistance and toxicity to two sucking pests, the whitefly, Bemisia tabaci, and the rice brown planthopper, Nilaparvata lugens, respectively. Moreover, in the field, transgenic rice plants expressing AaIT/GNA exhibited a significant improvement in grain yield when infested with N. lugens. This study shows that expressing the AaIT/GNA fusion protein in transgenic plants can be a useful approach for controlling pests, particularly sucking pests which are not susceptible to the toxin in Bt crops.
Assuntos
Antibiose , Arabidopsis/fisiologia , Herbivoria/efeitos dos fármacos , Insetos/fisiologia , Nicotiana/fisiologia , Oryza/fisiologia , Venenos de Escorpião/farmacologia , Animais , Arabidopsis/genética , Galanthus/química , Hemípteros/crescimento & desenvolvimento , Hemípteros/fisiologia , Insetos/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Larva/fisiologia , Lectinas de Ligação a Manose/genética , Lectinas de Ligação a Manose/farmacologia , Mariposas/crescimento & desenvolvimento , Mariposas/fisiologia , Ninfa/crescimento & desenvolvimento , Ninfa/fisiologia , Oryza/genética , Lectinas de Plantas/genética , Lectinas de Plantas/farmacologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Venenos de Escorpião/genética , Escorpiões/química , Nicotiana/genéticaRESUMO
BACKGROUND: The whitefly Bemisia tabaci is an important agricultural pest with global distribution. This phloem-sap feeder harbors a primary symbiont, "Candidatus Portiera aleyrodidarum", which compensates for the deficient nutritional composition of its food sources, and a variety of secondary symbionts. Interestingly, all of these secondary symbionts are found in co-localization with the primary symbiont within the same bacteriocytes, which should favor the evolution of strong interactions between symbionts. RESULTS: In this paper, we analyzed the genome sequences of the primary symbiont Portiera and of the secondary symbiont Hamiltonella in the B. tabaci Mediterranean (MED) species in order to gain insight into the metabolic role of each symbiont in the biology of their host. The genome sequences of the uncultured symbionts Portiera and Hamiltonella were obtained from one single bacteriocyte of MED B. tabaci. As already reported, the genome of Portiera is highly reduced (357 kb), but has kept a number of genes encoding most essential amino-acids and carotenoids. On the other hand, Portiera lacks almost all the genes involved in the synthesis of vitamins and cofactors. Moreover, some pathways are incomplete, notably those involved in the synthesis of some essential amino-acids. Interestingly, the genome of Hamiltonella revealed that this secondary symbiont can not only provide vitamins and cofactors, but also complete the missing steps of some of the pathways of Portiera. In addition, some critical amino-acid biosynthetic genes are missing in the two symbiotic genomes, but analysis of whitefly transcriptome suggests that the missing steps may be performed by the whitefly itself or its microbiota. CONCLUSIONS: These data suggest that Portiera and Hamiltonella are not only complementary but could also be mutually dependent to provide a full complement of nutrients to their host. Altogether, these results illustrate how functional redundancies can lead to gene losses in the genomes of the different symbiotic partners, reinforcing their inter-dependency.
Assuntos
Enterobacteriaceae/genética , Genoma Bacteriano , Halomonadaceae/genética , Hemípteros/genética , Hemípteros/microbiologia , Simbiose/genética , Aminoácidos/biossíntese , Animais , DNA/análise , DNA/isolamento & purificação , DNA/metabolismo , Hemípteros/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Hibridização in Situ Fluorescente , Redes e Vias Metabólicas/genética , Dados de Sequência Molecular , Análise de Sequência de DNA , Vitaminas/biossínteseRESUMO
Superoxide dismutases (SODs) are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase (ecCuZnSOD) complementary DNA was cloned and characterized from the whitefly, Bemisia tabaci. Quantitative polymerase chain reaction analysis showed that the expression level of BtecCuZnSOD was more than 10-fold higher in the invasive Middle East Asia Minor 1 (MEAM1) than in the native Asia II 3 species of the B. tabaci species complex. After exposure to low temperature (4 °C), the expression of Bt-ecCuZnSOD gene was significantly up-regulated in MEAM1 but not in Asia II 3. Furthermore, the expression level of B. tabaci intracellular CuZnSOD (Bt-icCuZnSOD), Bt-ecCuZnSOD and mitochondrial MnSOD (Bt-mMnSOD) was compared after transferring MEAM1 and Asia II 3 whiteflies from favorable (cotton) to unfavorable host plants (tobacco). On cotton, both CuZnSOD genes were expressed at a higher level in MEAM1 compared with Asia II 3. Interestingly, after transferring onto tobacco, the expression of Bt-ecCuZnSOD was significantly induced in Asia II 3 but not in MEAM1. On the other hand, while Bt-mMnSOD was expressed equally in both species on cotton, Bt-mMnSOD messenger RNA was up-regulated in MEAM1 on tobacco. Consistently, enzymatic activity assays of CuZnSOD and MnSOD demonstrated that CuZnSOD might play an important protective role against oxidative stress in Asia II 3, whereas MnSOD activation was critical for MEAM1 whiteflies during host adaptation. Taken together, our results suggest that the successful invasion of MEAM1 is correlated with its constitutive high activity of CuZnSOD and inducible expression of MnSOD under stress conditions.
Assuntos
Hemípteros/enzimologia , Superóxido Dismutase/genética , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar , Expressão Gênica , Gossypium/parasitologia , Hemípteros/genética , Interações Hospedeiro-Parasita , Espécies Introduzidas , Dados de Sequência Molecular , Estresse Oxidativo , RNA Mensageiro , Reação em Cadeia da Polimerase em Tempo Real , Superóxido Dismutase/isolamento & purificação , Superóxido Dismutase/metabolismo , Temperatura , Nicotiana/parasitologiaRESUMO
In phytophagous insects, experience can increase positive responses towards non-host plant extracts or induce oviposition on non-host plants, but the underlying chemical and behavioral mechanisms are poorly understood. By using the diamondback moth, Plutella xylostella, its host plant Chinese cabbage, and a non-host plant Chrysanthemum morifolium, as a model system, we observed the experience-altered olfactory responses of ovipositing females towards volatiles of the non-host plant, volatiles of pure chemicals (p-cymene and alpha-terpinene) found in the non-host plant, and volatiles of host plants treated with these chemicals. We assessed the experience-altered oviposition preference towards host plants treated with p-cymene. Naive females showed aversion to the odors of the non-host plant, the pure chemicals, and the pure chemical-treated host plants. In contrast, experienced females either became attracted by these non-host odors or were no longer repelled by these odors. Similarly, naive females laid a significantly lower proportion of eggs on pure chemical-treated host plants than on untreated host plants, but experienced females laid a similar or higher proportion of eggs on pure chemical-treated host plants compared to untreated host plants. Chemical analysis indicated that application of the non-host pure chemicals on Chinese cabbage induced emissions of volatiles by this host plant. We conclude that induced preference for previously repellent compounds is a major mechanism that leads to behavioral changes of this moth towards non-host plants or their extracts.
Assuntos
Brassica/parasitologia , Chrysanthemum/parasitologia , Monoterpenos/farmacologia , Mariposas/efeitos dos fármacos , Feromônios/farmacologia , Animais , Brassica/química , Chrysanthemum/química , Monoterpenos Cicloexânicos , Cimenos , Comportamento Alimentar , Feminino , Interações Hospedeiro-Parasita , Monoterpenos/análise , Mariposas/fisiologia , Odorantes , Oviposição , Feromônios/análiseRESUMO
Botanical preparations, usually from non-host plants, can be used to manipulate the behaviour of insect pests and their natural enemies. In this study, the effects of extracts of Chrysanthemum morifolium, a non-host plant of the diamondback moth, Plutella xylostella (Linnaeus), on the olfactory and oviposition responses of this phytophagous insect and on levels of parasitism by its specialist parasitoid Cotesia plutellae (Kurdjumov) were examined, using Chinese cabbage Brassica campestris L. ssp. pekinensis as the test host plant. Olfactometer tests showed that volatiles of chrysanthemum extract-treated host plants were less attractive to P. xylostella females than those from untreated host plants; and in contrast, volatiles of the chrysanthemum extract-treated host plants were more attractive to females of its parasitoid C. plutellae than those from untreated host plants. Oviposition preference tests showed that P. xylostella females laid only a small proportion of their eggs on chrysanthemum extract-treated host plants, while ovipositing parasitoid females parasitized a much higher proportion of host larvae feeding on the treated host plants than on untreated host plants. These results suggest that certain non-host plant compounds, when applied onto a host plant, may render the plant less attractive to a phytophagous insect but more attractive to its parasitoids. Application of such non-host plant compounds can be explored to develop push-pull systems to reduce oviposition by a pest insect and at the same time enhance parasitism by its parasitoids in crops.
Assuntos
Himenópteros/fisiologia , Lepidópteros/efeitos dos fármacos , Lepidópteros/parasitologia , Oviposição/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Brassica/parasitologia , Chrysanthemum/química , Feminino , Interações Hospedeiro-Parasita , Repelentes de Insetos/química , Repelentes de Insetos/farmacologia , Larva/efeitos dos fármacos , Larva/parasitologia , Extratos Vegetais/químicaRESUMO
The oviposition responses of the diamondback moth, Plutella xylostella L., to Neemix 4.5, a neem-based oviposition repellent/deterrent, with or without previous experience were studied in the laboratory on plain aluminum foil sheets, on aluminum foil sheets coated with cabbage juice, and on cabbage plants. In the plain aluminum foil sheet experiment, the females without prior experience of Neemix (inexperienced females) deposited more eggs (66.2%) on sections with untreated sheets than on sections with Neemix-treated sheets (33.8%), indicating that the inexperienced females were significantly repelled and/or deterred by Neemix. In contrast, the Neemix-experienced females deposited more eggs (69.2-69.7%) on sections with Neemix-treated sheets than on sections with untreated sheets (30.3-30.8%), implying that the females were not repelled and/or deterred by Neemix following an experience. In the tests using egg-laying sheets coated with cabbage juice or with cabbage juice plus Neemix, the inexperienced females deposited more eggs (63.8%) on the sections with sheets treated with cabbage juice only than on the sections with sheets coated with cabbage juice and Neemix (36.2%), indicating that inexperienced females were repelled and/or deterred by Neemix. Similarly, Neemix-experienced females deposited more eggs (62.2-65.6%) on the sections with sheets treated with cabbage juice plus Neemix than on the sections with sheets treated with juice only (34.4-37.8%), suggesting that the females were not repelled and/or deterred by Neemix following an experience. On cabbage plants, inexperienced females oviposited similar numbers of eggs on both Neemix-treated (50.8%) and untreated plants (49.2%). However, the Neemix-experienced females oviposited more eggs on the plants treated with Neemix (56.3%) than on untreated plants (43.7%), indicating that the females were attracted by Neemix-treated plants following an experience. The significance of this study for applications of insect repellents/deterrents in pest management is discussed.