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1.
Western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), cause very large economic damage on a variety of field and greenhouse crops. In this study, plant resistance against thrips was introduced into transgenic potato plants through the expression of novel, custom-made, multidomain protease inhibitors. Representative classes of inhibitors of cysteine and aspartic proteases [kininogen domain 3 (K), stefin A (A), cystatin C (C), potato cystatin (P) and equistatin (EIM)] were fused into reading frames consisting of four (K-A-C-P) to five (EIM-K-A-C-P) proteins, and were shown to fold into functional inhibitors in the yeast Pichia pastoris. The multidomain proteins were expressed in potato and found to be more resistant to degradation by plant proteases than the individual domains. In a time span of 14-16 days, transgenic potato plants expressing EIMKACP and KACP at a similar concentration reduced the number of larvae and adults to less than 20% of the control. Leaf damage on protected plants was minimal. Engineered multidomain cysteine protease inhibitors thus provide a novel way of controlling western flower thrips in greenhouse and field crops, and open up possibilities for novel insect resistance applications in transgenic crops.  相似文献   

2.
Proteolytic activity in whole insect extracts of the western flower thrips, Frankliniella occidentalis, was found to belong predominantly to the class of cysteine proteases. The pH optimum of the general proteolytic activity was determined to be 3.5, which is low when compared to other insects using cysteine proteases for protein digestion. The proteinaceous cysteine protease inhibitors chicken cystatin, potato cystatin and sea anemone equistatin inhibited in vitro more than 90% of the protease activity. To test in vivo the biological effect of such inhibitors on the oviposition rate of western flower thrips, recombinant potato cystatin and equistatin were fed to adult females. A gradual reduction in oviposition rate to about 45% of control was observed when reared on these PIs for a period of 5 days, with no increase in mortality. These results are discussed in the light of the application of protease inhibitors in transgenic plants to control this insect pest.  相似文献   

3.
The spread of the western flower thrips Frankliniella occidentalis (Pergande)   总被引:17,自引:0,他引:17  
Abstract 1 Since the late 1970s, the western flower thrips has spread from its original distribution in western North America to become a major worldwide crop pest. 2 A wide range of data sources have been used to map the original distribution in the U.S.A. and Canada, and the progress of the spread in the U.S.A., Canada, Europe, northern Africa and Australia. 3 The possible reasons for the start of the spread are discussed. The most likely reason is that intensive insecticide use in horticulture in the 1970s and 1980s selected an insecticide resistant strain or strains. These then established in glasshouses across North America and spread from there to Europe, Asia, Africa and Australia. 4 The international spread of the western flower thrips occurred predominantly by the movement of horticultural material, such as cuttings, seedlings and potted plants. Within Europe, an outward spread from the original outbreak in the Netherlands is discernible. The speed of spread was 229 ± 20 km/year. 5 The spread has not been restricted to glasshouses. The western flower thrips has established outdoors in areas with milder winters; for example, across the southern U.S.A., southern Europe and Australia. It also overwinters in some regions with colder winters. 6 Polyphagous phytophagous thrips have many factors predisposing them to become worldwide crop pests, particularly in glasshouses. Some other species that might spread in a similar way to the western flower thrips are listed.  相似文献   

4.
In this study, the effects of the accumulation of cysteine protease inhibitors on the food preferences of adult female western flower thrips, Frankliniella occidentalis (Pergande), were investigated. Representative members of the cystatin and thyropin gene families (stefin A, cystatin C, kininogen domain 3 and equistatin) were expressed in potato (Solanum tuberosum) cv. Impala, Kondor and Line V plants. In choice assays, a strong time- and concentration-dependent deterrence from plants expressing stefin A and equistatin was observed. Cystatin C and kininogen domain 3 were not found to be active. All tested inhibitors were equally or more active than stefin A at inhibiting the proteolytic activity of thrips, but, in contrast with stefin A, they were all expressed in potato as partially degraded proteins. The resistance of cysteine protease inhibitors against degradation in planta by endogenous plant proteases may therefore be relevant in explaining the observed differences in the deterrence of thrips. The results demonstrate that, when given a choice, western flower thrips will select plants with low levels of certain cysteine protease inhibitors. The novel implications of the defensive role of plant cysteine protease inhibitors as both deterrents and antimetabolic proteins are discussed.  相似文献   

5.
Highly polymorphic microsatellite markers can provide important demographic information on founder events and range expansion following initial introduction of invasive insect species. Six microsatellite loci were isolated from an enriched DNA library in order to study the invasion patterns of the western flower thrips, Frankliniella occidentalis. All loci tested were found to be polymorphic and successfully amplified in all individuals. The number of alleles per locus ranged from five to nine and heterozygosity ranged from 45 to 73%. Some of the loci were also successfully amplified in other thrips species.  相似文献   

6.
A rapid cold hardening process is reported in first instar larvae of Frankliniella occidentalis. When larvae are transferred directly from 20 degrees C to -11.5 degrees C for 2h there is 78% mortality, whereas exposure to 0 degrees C for 4h prior to transfer to -11.5 degrees C reduces mortality to 10%. The response can also be induced by exposure to 5 degrees C for 4h or by gradual cooling at rates between 0.1 and 0.5 degrees C min(-1.) The acquired cold tolerance is transient and is rapidly lost (after 1h at 20 degrees C). Rapid cold hardening extends survival times at -11.5 degrees C and depresses lethal temperatures in short (2h) exposures. Rearing at 15 degrees C (12L:12D), (a cold acclimation regime for F. occidentalis), does not protect against the cold shock induced by direct transfer to -11.5 degrees C (which rapid cold hardening does) but does extend survival time at -5 degrees C (i.e. increased chill tolerance) whilst rapid cold hardening does not. The rapid and longer term cold hardening responses in F. occidentalis therefore appear to have different underlying mechanisms.  相似文献   

7.
Philasterides dicentrarchi is a causative agent of scuticociliatosis in olive flounder Paralichthys olivaceus, aquaculture in Korea. In this study, a cDNA encoding a cathepsin L-like cysteine protease (PdCtL) of P. dicentrarchi (synonym Miamiensis avidus) was identified. To express the PdCtL recombinant protein in a heterologous system, 10 codons were redesigned to conform to the standard eukaryotic genetic code using polymerase chain reaction (PCR)-based site-directed mutagenesis. The recombinant P. dicentrarchi procathepsin L (proPdCtL) was expressed at high levels in E. coli Rosetta (DE3) pLysS with a pPET21a vector, and successfully refolded, purified, and activated into a functional and enzymatically active form. The optimal pH for protease activity was 5. Similar to other cysteine proteases, enzyme activity was inhibited by E64 and leupeptin. Immunogenicity of recombinant PdCtL was assessed by enzyme-linked immunosorbent assay, western blot, and specific anti-recombinant PdCtL antibodies were detected. Our results suggest that the biochemical characteristics of the recombinant ciliate proPdCtL protein are similar to those of the cathepsin L-like cysteine protease, that the PCR-based site-direct mutated ciliate gene was successfully expressed in a biochemically active form, and that the recombinant PdCtL acted as a specific epitope in olive flounder.  相似文献   

8.
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10.
西花蓟马Frankliniella occidentalis(Pergande)是世界性的园艺作物上的重要害虫,几乎对每种类型的杀虫剂均产生了抗药性,包括有机磷、氨基甲酸酯、拟除虫菊酯类杀虫剂和多杀菌素等。本文对国外西花蓟马的抗药性发展现状和抗性机制进行了总结,并提出了抗性治理策略,即科学合理使用杀虫剂,结合栽培防治、物理防治、生物防治和寄主植物抗性等方式降低杀虫剂对西花蓟马的选择压,从而达到抗性治理的目的。  相似文献   

11.
All life stages of the western flower thrips (Frankliniella occidentalis Pergande) were tested for their response to the fumigant methyl bromide and dose response curves were established. Pseudo-pupae were the most tolerant stage with a calculated LD99 of 53.1 mg litre“1 h at 15oC with a gas concentration of 13.5 mg litre”1, but no survivors were found in any test above a concentration time product of 40 mg litre“1 h. Standard 4 h fumigations achieving a CTP of 54 mg litre”1 h that are used to control other pests are therefore likely to give very high levels of control of western flower thrips.  相似文献   

12.
The systemic effects of neem on the western flower thrips, Frankliniella occidentalis (Pergande), were investigated in laboratory trials using green bean, Phaseolus vulgaris L., in arena and microcosm experiments. In arena experiments, systemic effects of neem against western flower thrips larvae on primary bean leaves were observed with maximum corrected mortality of 50.6%. In microcosm experiments using bean seedlings, higher efficacy in the control of western flower thrips were observed with soil applications of neem on a substrate mixture (i.e., Fruhstorfer Erde, Type P, and sand) in a 1:1 ratio (93% corrected mortality) compared with application on the commercial substrate only (76% corrected mortality). However, longer persistence of neem was observed with soil application on the commercial substrate, which showed effects against thrips for up to 6 d after application. In addition to systemic effects observed on all foliage-feeding stages of western flower thrips, mortality on contact and repellent effects were observed on soil-inhabiting stages after soil applications of neem. Finally, bean seedlings grown from seeds pregerminated for 3 d in neem emulsion were also toxic to western flower thrips.  相似文献   

13.
Aims: Larval stages of Frankliniella occidentalis are known to be refractory to fungal infection compared with the adult stage. The objective of this study was to identify promising fungal isolate(s) for the control of larval stages of F. occidentalis. Methods and Results: Ten isolates of Metarhizium anisopliae and eight of Beauveria bassiana were screened for virulence against second‐instar larvae of F. occidentalis. Conidial production and genetic polymorphism were also investigated. Metarhizium anisopliae isolates ICIPE 7, ICIPE 20, ICIPE 69 and ICIPE 665 had the shortest LT50 values of 8·0–8·9 days. ICIPE 69, ICIPE 7 and ICIPE 20 had the lowest LC50 values of 1·1 × 107, 2·0 × 107 and 3·0 × 107 conidia ml?1, respectively. Metarhizium anisopliae isolate ICIPE 69 produced significantly more conidia than M. anisopliae isolates ICIPE 7 and ICIPE 20. Internally transcribed spacers sequences alignment showed differences in nucleotides composition, which can partly explain differences in virulence. Conclusion: These results coupled with the previous ones on virulence and field efficacy against other species of thrips make M. anisopliae isolate ICIPE 69 a good candidate. Significance and Impact of the Study: Metarhizium anisopliae isolate ICIPE 69 can be suggested for development as fungus‐based biopesticide for thrips management.  相似文献   

14.
The impact of flowers on the population growth of Frankliniella occidentalis was characterized on waxflower plants, Impatiens walleriana . In the presence of flowers, the populations grew exponentially while in their absence, the thrips populations only maintained themselves and showed no growth The difference in number and in size of thrips reared in both conditions suggested that the flowers were essential for the success of thrips development. In the presence of flowers, the distribution of females between the leaves and the flowers varied strongly according to the availability of flowers. By contrast, the proportion of first larvae in the flowers increased up to the last day of the experiment.  相似文献   

15.
In insect reverse genetics, dietary delivery of interfering RNAs is a practical approach in nonmodel species, such as thrips, whose small size, and feeding behavior restricts the use of other delivery methods. In a laboratory context, an unsuitable diet could confound the interpretation of an RNA interference (RNAi) phenotype, however well-formulated artificial diets can minimize experimental variability, reduce the need for insect handling, and can further be used for roles, such as delivering double-strand RNA (dsRNA)-expressing recombinant bacteria. In this study, artificial diets for oral delivery of dsRNA were developed for two important pest thrips species, western flower thrips (Frankliniella occidentalis) and onion thrips (Thrips tabaci), with the goal of (a) stimulating feeding behavior, (b) supporting optimal growth rates of dsRNA-expressing symbiotic bacteria, and (c) nutritionally supporting the thrips for sufficient periods to observe RNAi phenotypes. The efficacy of artificial diets for ingesting “naked” dsRNA or dsRNA-expressing symbionts and dsRNA delivery via host plant uptake was evaluated. Compared with previously published diet formulations, new combinations based on tryptone, yeast, and soy were superior for enhancing feeding and longevity. However, simply adding “naked” dsRNA to an artificial diet was an unreliable form of RNAi delivery in our hands due to dsRNA degradation. Delivery via host plants was more successful, and the new diet formulation was suitable for symbiont-mediated dsRNA delivery, which we believe is the most convenient approach for large-scale knockdown experiments. This study, therefore, provides alternative methodologies for thrips rearing, dietary RNAi delivery, and insights into the challenges of performing dietary RNAi in nonmodel insects.  相似文献   

16.
The general principles in pathogen transmission by insects involve a complex and specific interplay, in this case between thrips, tospovirus and their shared host plant, which has led to outbreaks of crop disease epidemics of economic and social importance. The possible processes and factors driving their co‐evolution were partly studied by rearing Frankliniella occidentalis [western flower thrips (WFT)] on either tomato spotted wilt virus (TSWV)–infected or uninfected Capsicum annum leaflets throughout their larval stages. Later, pupae were transferred individually on healthy leaf discs for further studies of the influence of TSWV on WFT development and behavioural patterns. The exposure of WFT to TSWV was found to improve performance with regard to longevity and survival, with mean longevity being significantly higher in TSWV‐exposed WFT compared to unexposed ones (F(3,403) = 22.44, P < 0.0001). The observed improvement in survival was as a result of significant reduction in mortality for the WFT individuals exposed to TSWV (F(3,383) = 849.94, P < 0.0001) compared to the unexposed. However, the results showed a significant reduction in mean daily fecundity overtime (F10,10) = 246.66, P < 0.0001) and across the four treatments (F(3,30) = 6.62, P = 0.001), as well as lifetime fecundity (F(3,23) = 21.23, P < 0.0001) of the WFT exposed to TSWV compared to the unexposed reared on uninfected leaf discs. For preferential test, C. annum leaf discs infected with TSWV were more attractive to WFT as compared to healthy leaf discs (χ2(4, 34) = 112.35, P < 0.0001). These results are envisaged to contribute to a clear understanding into the plant–vector–virus interaction, which is essential for accurate diagnosis and control of the TSWV epidemic, as well as the control of F. occidentalis as crop pest.  相似文献   

17.
There is limited information on the biology and pathogenesis of Leishmania aethiopica, causative agent of cutaneous leishmaniasis (CL) in Ethiopia. In this study we have identified and characterized two cathepsin L-like cysteine protease genes, Laecpa and Laecpb, from L. aethiopica. The predicted amino acid sequence of Laecpa and Laecpb is more than 75% identical with homologous cathepsin L-like cysteine protease genes of other Leishmania species and less than 50% identical with human cathepsin L. Laecpa is expressed predominantly in the stationary, and to a lower level, during the amastigote stage while Laecpb is specifically expressed in the stationary stage of L. aethiopica development. Phylogenetic analysis showed that the two genes are grouped into separate clades which are the result of gene duplication. The isolation of these genes will be useful in developing Leishmania species specific diagnostics for molecular epidemiological studies and serves as a first step to study the role of cysteine proteases in L. aethiopica pathogenesis.  相似文献   

18.
Western flower thrips (Frankliniella occidentalis) develops from an active larval stage through to a non-feeding, almost immobile, pre-pupal and pupal stage. This generally occurs in the compost or soil below the plant on which the larvae fed. Control of thrips at this stage in their development offers a chance of utilising pathogens or pesticides not suitable for use in an integrated control programme aimed at adult and larval stages. Trials were done with F. occidentalis pupae and pre-pupae in a soil/peat based compost using 11 pesticides, three fungal pathogens and four species of insect parasitic nematodes. The pesticides malathion and chlorpyrifos-methyl gave the most promising result with 97.5% and 96.5% control, respectively. The fungal pathogen Metarhizium anisopliae proved better when applied as a pre-pupation rather than as a post-pupation treatment (74.5% : 26.6% control). The insect parasitic nematode Steinernema carpocapsae gave 76.6% control when applied at 25 times 104 nematodes litre-1 of compost. The results are discussed in relation to control of thrips in glasshouses.  相似文献   

19.
Western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), is a major pest of horticultural crops worldwide. The search for alternative pest management techniques has resulted in increasing interest in the use of kairomones and other behaviour‐modifying chemicals to mitigate the impact of this pest. In this study, we determined whether the origin of populations, feeding history, and/or genotype influence the response of WFT to the thrips kairomone lure methyl isonicotinate (MI) in a Y‐tube olfactometer study. Four New Zealand thrips populations were tested: (1) from a commercial glasshouse capsicum crop, (2) from a long‐established laboratory colony (>222 generations) kept on chrysanthemums, (3) from a laboratory colony (6–9 generations) kept on French dwarf beans, and (4) thought to be a separate cryptic non‐pest species from outdoor yellow tree lupins, Lupinus arboreus Sims (Fabaceae). In the laboratory tests, significantly more WFT from all four populations chose the MI‐laden arm of a Y‐tube olfactometer when it contained 1 μl methyl isonicotinate (61.3–73.2%) compared with the blank no‐odour arm. No differences in response to MI were found between the two laboratory and the one glasshouse WFT populations. Both laboratory populations and the greenhouse population belonged to the ‘glasshouse pest’ genotype of WFT. However, the cryptic non‐pest WFT genotype responded more strongly to MI than any of the other populations, although the response was only significantly stronger than that of the long‐established laboratory population. Significant differences were also found among populations in the average time taken for thrips to make a choice to enter either arm of the Y‐tube olfactometer, with the cryptic non‐pest lupin genotype taking the shortest time, followed by thrips from the capsicum glasshouse. The results are discussed with respect to the variability in olfactory perception and olfactory behaviour within a species and the relevance to the use of such a kairomone lure in pest management programmes.  相似文献   

20.
刘丽辉  张帆  吴珍泉 《生态学报》2008,28(10):4891-4895
在15、20、25℃和30℃,RH75%的条件下,分析了温度与西花蓟马发育速率的关系,并测定了西花蓟马的发育起点温度、有效积温和存活率。结果表明:在15~30℃范围内,西花蓟马各虫期的发育速率随温度的升高而加快;用直接最优法对发育起点温度和有效积温进行估算,求得西花蓟马的卵、若虫、预蛹和伪蛹及世代发育起点温度分别为5.70、7.04、4.49、6.84℃和6.23℃;世代的有效积温为219.73d&#183;℃。西花蓟马的世代存活率在25℃时最高,为40.12%;30℃时西花蓟马的存活率最低,为17.80%。  相似文献   

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