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1.
Taro Colocasia esculenta (L. Schott) and rice (Oryza sativa L.) form a major part of the staple diet of pacific islanders. Pest constraints hamper the sustainability of taro and rice production in the Pacific region. Insect feeding trials were conducted in vitro to determine the effects of plant lectins against planthopper pests of taro and rice. Lectins were incorporated into artificial diet at 0.1% (w/v) level. The lectins Galanthus nivalis agglutinin (GNA) and concanavalin A (Con A) showed significant antimetabolic effects towards third instar nymphs of taro planthopper (Tarophagous proserpina Kirkaldy) whilst Pisum sativum agglutinin (PSA) showed no significant effects toward the insect. Psophocarpus tetragonolobus agglutinin (PTA) showed significant antimetabolic effects towards third instar nymphs of rice brown planthopper (Nilaparvata lugens Stål). PTA also reduced honeydew excretion levels of rice brown planthopper, over a 24-hour period, demonstrating antifeedant properties of the protein.  相似文献   

2.
Insect feeding trials were carried out to determine the effects of incorporating a range of plant derived proteins into artificial diets fed to leafhopper and planthopper pests of rice. The lectins Galanthus nivalis agglutinin (GNA) and wheat germ agglutinin (WGA), and the enzyme soy bean lipoxygenase (LPO) were shown to exhibit significant antimetabolic effects towards first and third instar nymphs of rice brown planthopper (Nilaparvata lugens Stål) when incorporated into artificial diet at 0.1% (w/v), 0.1% (w/v) and 0.08% (w/v) levels respectively. The lectin GNA was also shown to exhibit a significant antimetabolic effect towards third instar nymphs of the rice green leafhopper (Nephotettix cinciteps Uhler). A number of inert proteins, lectins, protein inhibitors and enzymes also tested showed relatively little or no effect towards both insects.  相似文献   

3.
Insect feeding trials were carried out to determine the effects of a range of mannose-specific lectins on third instar nymphs of the rice brown planthopper (BPH), Nilaparvata lugens. Stål. Dose response curves show that Galanthus nivalis agglutinin (GNA) has the strongest toxic effect of the lectins tested, and is effective at concentrations considerably lower than those previously reported. Narcissus pseudonarcissus agglutinin (NPA) and Allium sativum agglutinin (ASA) exhibit a significant antimetabolic effect towards the insect but were less effective (on a molar basis) than GNA. LC50 values for GNA, NPA and ASA are approximately 4 μM, 11 μM and >40 μM respectively. These mannose-specific lectins are serologically identical, but differ in the number of subunits per protein molecule; ASA is a dimer, NPA is a trimer and GNA is a tetramer. The results obtained support the hypothesis, that the effectiveness of the mannose-binding lectins as antimetabolites is determined by the number of subunits per molecule. Two N-acetylglucosamine binding lectins, the dimeric Oryza sativa agglutinin (OSA) and the monomeric Urtica dioica agglutinin (UDA), were also tested but at a concentration of 0.1% w/v exhibited no significant antimetabolic effect towards BPH, although the related lectin wheatgerm agglutinin (WGA) has previously been demonstrated to be toxic towards the insect.  相似文献   

4.
Five transgenic rice lines, each containing an insecticidal toxin gene from Bacillus thuringiensis (Bt) under control of a different promoter, were tested for effects on two non-target insects: the brown planthopper, Nilaparvata lugens (Stål) (Homoptera: Delphacidae), and its predator Cyrtorhinus lividipennis (Hemiptera: Miridae). Bt toxin was detected by ELISA in the honeydew of N. lugens that fed on rice lines with the CaMV 35S and actin promoters. Nilaparvata lugens produced greater volumes of acidic honeydew (derived from xylem feeding) on all five Bt rice lines than on non-transgenic control lines. The amount of honeydew derived from phloem feeding did not differ between Bt and control lines. There were no differences between N. lugens reared on Bt and control lines in any of the five fitness parameters measured (survival to the adult stage, male and female weight, and male and female developmental time). There were no differences between C. lividipennis reared on N. lugens nymphs from Bt and control lines, in any of the three fitness parameters examined (survival to the adult stage and male and female developmental time). Our results indicate that N. lugens and its natural enemies will be exposed to Bt toxins from rice lines transformed with some Bt gene constructs, but that this exposure might not affect N. lugens and C. lividipennis fitness.  相似文献   

5.
Summary Mature seed-derived callus from an elite Chinese japonica rice cv. Ewan 5 was cotransformed with two plasmids, pWRG1515 and pRSSGNAl, containing the selectable marker hygromycin phosphotransferase gene (hpt), the reporter β-glucuronidase gene (gusA) and the snowdrop (Galanthus nivalis) lectin gene (gna) via particle bombardment. Thirty-five independent transgenic rice plants were regenerated from 177 bombarded calluses. Eighty-three percent of the transgenic plants contained all three genes, as revealed by Southern blot analysis. Western blot analysis revealed that 23 out of 29 gna-containing transgenic plants expressed Galanthus nivalis agglutinin (GNA) (79%) at various levels, with the highest expression being approximately 0.5% of total soluble protein. Genetic analysis confirmed Mendelian segregation of all three transgenes (gna, hpt and gusA) in the R2 progeny. Amongst the R2 generation two independent homozygous lines were identified that expressed all three transgenes. Insect bioassay and feeding tests showed that these homozygous lines had significant inhibition to rice brown planthopper (Nilaparvata lugens, BPH) by decreasing the survival, overall fecundity of BPH, retarding development, and decreasing the feeding of BPH. These BPH-resistant lines have been incorporated into a rice insect resistance breeding program. This is the first report that homozygous transgenic rice lines expressing GNA, developed by genetic transformation and through genetic analysis-based selection, conferred enhanced resistance to BPH.  相似文献   

6.
In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through theAgrobacterium tumefaciens- mediated method. The toxicity of GNA-expressing plants to aphids has also been studied. The independently derived plants were subjected to molecular analyses. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that thegna gene was integrated into maize genome and inherited to the following generations. The typical Mendelian patterns of inheritance occurred in most cases. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of nine GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to aphids. These plants synthesized GNA at levels above 0.22% total soluble protein, and enhanced resistance to aphids was demonstrated by exposing the plants to corn leaf aphid (Rhopalosiphum maidis Fitch) under greenhouse conditions. The nymph production was significantly reduced by 46.9% on GNA-expressing plants. Field evaluation of the transgenic plants supported the results from the inoculation trial. After a series of artificial self-crosses, some homozygous transgenic maize lines expressing GNA were obtained. In the present study, we have obtained new insect-resistant maize material for further breeding work.  相似文献   

7.
We used particle bombardment to transform two elite Thai rice varieties, Khao Dawk Mali 105 (KDML105) and Supanburi 60 (SP60), with the snowdrop lectin gene gna (Galanthus nivalis agglutinin). This gene confers resistance to sap-sucking insects such as the brown planthopper (BPH; Nilaparvata lugens), which is one of the most damaging pests of rice. Traditionally, KDML105 and SP60 have been regarded as recalcitrant to transformation, and this is the first account of successful gene transfer to these varieties. By molecular analysis, we confirmed the recovery of over thirty gna-transgenic lines. GNA protein expression was characterised by western blot analysis, and we achieved expression levels of up to 0.25% total soluble protein. GNA-producing R1 transgenic plants were significantly more resistant to BPH than control plants (P<0.0001), with 37% and 42% reduction in nymphal survival for constitutive and phloem-specific expression, respectively. Transferring the gna gene to these superior rice varieties thus represents a major step forward for crop improvement in Thailand, and should help to reduce the damage caused by rice pests, and hence increase yields for this vital domestic and export market.  相似文献   

8.
1 Insecticidal proteins can be excreted in the honeydew when sap-sucking insects feed on insect-resistant transgenic plants. Honeydew can be an important source of carbohydrates, thus potentially exposing a broad range of honeydew-feeding insects to transgene products.
2 Snowdrop lectin ( Galanthus nivalis agglutinin; GNA) dissolved in a 2 m sucrose solution had no antifeedant effect on female aphid parasitoids ( Aphidius ervi ) but had a direct negative effect on their longevity.
3 When feeding on honeydew from Rhopalosiphum padi feeding on a GNA-containing artificial diet, Aphidius ervi suffered a longevity reduction that was more pronounced than was to be expected based on the detected GNA concentration in the honeydew.
4 Analysis of carbohydrate and amino acid composition revealed that a change in honeydew composition caused by a GNA-effect on the aphids could be a possible explanation for the additional reduction in parasitoid longevity.
5 When comparing the effect of honeydew from Sitobion avenae and R. padi feeding on GNA-expressing or nontransformed wheat plants on A. ervi longevity, aphid species was found to have a significant effect, whereas the wheat variety had no effect. The latter result was probably due to low GNA expression levels in the plants. Differences in nutritional suitability between honeydew from R. padi and S. avenae could be explained by differences in carbohydrate and amino acid composition.
6 This is the first study to demonstrate that GNA ingested by aphids and transported into the honeydew can negatively affect the parasitoids consuming this honeydew.
7 We recommend that honeydew should be considered as a route of exposure to transgene products in future risk assessment studies.  相似文献   

9.
Transgenic rice plants expressing snowdrop lectin (Galanthus nivalis agglutinin; GNA) were screened for resistance to green leafhopper (Nephotettix virescens; GLH), a major homopteran pest of rice. Survival was reduced by 29% and 53% (P<0.05) respectively, on plants where GNA expression was tissue-specific (phloem and epidermal layer) or constitutive. Similar levels of resistance in GNA-expressing transgenic rice were previously reported for rice brown planthopper (Nilaparvata lugens; BPH). GNA binding to glycoproteins in gut tissues showed that BPH contained more "receptors" than GLH, and that the binding affinity was stronger, particularly in the midgut. Subsequent toxicity of GNA is thus unlikely to be directly related to the amount of lectin bound. GNA was not detected in the honeydew of either insect species when they were fed on GNA-expressing plants, in contrast to results from artificial diet studies. This result suggests that GNA is not being delivered to the insect efficiently. When offered a free choice vs control plants, BPH nymphs tended to avoid plants expressing GNA; avoidance was less pronounced and took longer to develop on plants where GNA expression was tissue-specific, In contrast to BPH, GLH nymphs were attracted to plants expressing GNA, whether constitutively or in a tissue-specific manner.  相似文献   

10.
The gene coding for agglutinin from Galanthus nivalis (GNA) was expressed in, and secreted by, the methylotrophic yeast, Pichia pastoris. Transformants of P. pastoris were selected and a process to produce and purify gram quantities of recombinant GNA was developed. GNA was secreted at approximately 80 mg l–1 at the 200 l scale and was purified to 95% homogeneity using hydrophobic interaction chromatography. The recombinant protein was similar to the protein synthesised in plant with respect to structure and biological activity.  相似文献   

11.
A kairomone produced by the rice white-backed planthopper Sogatella furcifera (Horvath) for the mymarid egg parasitoid Anagrus nilaparvatae Pang et Wang was investigated. Eggs, female and male adults, nymphs, exuvia, honeydew, nymph-damaged plants and plants with S. furcifera eggs all elicited searching behavior in the parasitoid. Eggs, female adults and plants with eggs were the most attractive, while exuvia and honeydew evoked the weakest responses. The active compound(s) from S. furcifera female adults, nymphs and plants with eggs could be effectively extracted with methanol, acetone, n-hexane, ethyl ether and dichloromethane. Isolation of the active chemical(s) in three solvent extracts, the acetone and n-hexane extracts of S. furcifera female adults, and the acetone extract of S. furcifera nymphs, was accomplished by thin-layer chromatography. Only one fraction was active and had the same chemical properties in the three solvent extracts. Infra-red analysis of this fraction revealed a kind of ester with saturated and unsaturated fatty acids, which was previously found to be active in the same compound as the extracts from the nymphs and female adults of the rice brown planthopper, Nilaparvata lugens (Stål), and identified as palm oil. The results are discussed in relation to host location by A. nilaparvatae.  相似文献   

12.
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.  相似文献   

13.
傅强  张志涛胡萃 《昆虫学报》1997,40(-1):116-121
对寄主经历不同的稻褐飞虱Nilaparvata lugens 3龄若虫致害特性的研究结果表明:在抗虫品种Mudgo上的生活经历显著提高试虫在含Bphl基因的Mudeo、IR26、IR28、IR34上的蜜露排泄量、存活率及试虫对Mudgo的选择性:而ASD7上的经历仅显著提高试虫在ASD7上的蜜露量和存活率。不同品种上的经历对试虫的影响有所不同。同时,各经历试虫 在抗虫品种上的蜜露排泄量显著小于在TNl上的排泄量。  相似文献   

14.
Abstract The current study investigated changes in the content of the rare earth element Europium (Eu) in roots, shoots and leaves of rice plants and in Nilaparvata lugens Stål (Hemiptera: Delphacidae) using an Eu marker and hydroponic culture. The results showed that N. lugens infestation significantly reduced Eu content in roots, shoots and leaves of two rice varieties, Shenyou 1 and Xieyou 963. The Eu content in roots, shoots and leaves of the susceptible variety, Shenyou 1, was significantly higher than that in the resistant variety, Xieyou 963. The Eu content of N. lugens fed on Shenyou 1 was significantly higher than of those fed on Xieyou 963. In addition, the Eu level was elevated at a higher density of N. lugens infestation. Eu content in the bodies of N. lugens was related to their weight and honeydew excretion, with a significant positive correlation. Thus, Eu content in the bodies of N. lugens can be considered an index of the amount of phloem sap taken in by N. lugens because the amount of honeydew excretion is proportional to the amount of phloem sap consumed. The ratios of Eu content in N. lugens to that in roots, shoots and leaves of rice plants were elevated at a higher N. lugens infestation density. That ratio was maximal in leaves, was intermediate in shoots and was minimal in roots. There was no significant difference in ratios between the two plant varieties. An Eu marker may be useful in the screening of resistant varieties and in the study of the mechanisms of resistance.  相似文献   

15.
Biolistic transformation was used to introduce genes encoding the insecticidal proteins snowdrop lectin (Galanthus nivalis agglutinin; GNA) and cry1Ac Bt toxin (-endotoxin from Bacillus thuringiensis) into elite rice (Oryza sativa) cultivars. Plant transformation was carried out in parallel experiments simultaneously by using either whole plasmids containing suitable gene constructs, or the corresponding minimal gene cassettes, which were linear DNA fragments lacking vector sequences excised from the plasmids. Both transformation methods generated similar numbers of independent transformation events. Selected R0 clonal plant lines were further characterised for presence and expression of transgenes. Co-transformation of the unselected genes (cry1Ac and gna) with the selectable marker (hpt) was at least as efficient for transformation with minimal gene cassettes as with whole plasmid DNA, and higher levels of accumulation of the insecticidal gene products GNA and cry1Ac were observed in plants resulting from minimal gene cassette transformation. Insect bioassays with major pests of rice showed that transgenic plants expressing gna showed enhanced resistance to brown planthopper (Nilaparvata lugens), and plants expressing cry1Ac were protected against attack by striped stem borer (Chilo suppressalis). Expression of both transgenes gave protection against both pests, but did not increase protection against either pest significantly over the levels observed in plants containing a single insecticidal transgene.  相似文献   

16.
Rice brown planthoppers (Nilaparvata lugens) were fed on artificial diet containing snowdrop lectin (Galanthus nivalis agglutinin; GNA), which has been shown to be toxic towards this insect pest. In addition to decreasing survival, the lectin affected development, reducing the growth rate of nymphs by approximately 50% when present at a concentration of 5.3&mgr;M. Immunolocalisation studies showed that lectin binding was concentrated on the luminal surface of the midgut epithelial cells within the planthopper, suggesting that GNA binds to cell surface carbohydrate moieties in the gut. Immunolabelling at a lower level was also observed in the fat bodies, the ovarioles, and throughout the haemolymph. These observations suggest that GNA is able to cross the midgut epithelial barrier, and pass into the insect's circulatory system, resulting in a systemic toxic effect. Electron microscope studies showed morphological changes in the midgut region of planthoppers fed on a toxic dose of GNA, with disruption of the microvilli brush border region. No significant proteolytic degradation of GNA was observed either in the gut or honeydew of planthoppers fed on lectin-containing diet. The presence of glycoproteins which bind GNA in the gut of the brown planthopper was confirmed using digoxigen-labeled lectins to probe blots of extracted gut polypeptides.  相似文献   

17.
The snowdrop lectin GNA (Galanthus nivalis agglutinin) has been shown to possess insecticidal activity to a range of economically important insect pests. However, the precise mechanism of insecticidal action of GNA against insects remains unknown. In this investigation, we attempted to purify and identify receptor(s) responsible for binding of GNA in the larval midgut of a major lepidopteran pest (the cotton leafworm, Spodoptera littoralis) to better understand its mode of action. Therefore, cytoplasmic as well as membrane proteins from 800 larval midguts were chromatographed on a column with immobilized GNA. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis analysis of the proteins eluted from the GNA column followed by sequencing of the GNA‐binding proteins and BLAST analyses revealed that the N‐terminal sequences of a 24 kDa polypeptide purified from the cytoplasmic and membrane protein fraction revealed sequence similarity to sequences encoding heavy chain homologs of ferritin from Manduca sexta (76% sequence identity), Calpodes ethlius (80% sequence identity) and Bombyx mori (61% sequence identity). Furthermore, the N‐terminal sequence of a 31 kDa polypeptide from the membrane protein fraction showed sequence similarity to a light chain homolog of ferritin from Manduca sexta (88% sequence identity).  相似文献   

18.
The feeding behaviour of Nilaparvata lugens (Stal.) was electronically monitored on three rice varieties showing different levels of resistance in the Philippines, over an 8 h period, using a DC amplifier and a video-assisted observation method to simultaneously record honeydew production.Six electrical penetration graph (EPG) patterns are described by their frequency and amplitude characteristics. Patterns 4 and 5 were associated with the production of two different types of honeydew; basic honeydew containing amino acids was produced during Pattern 4 and acidic honeydew with no amino acids was produced during Pattern 5. These patterns are thought to indicate phloem and xylem ingestion by N. lugens respectively.Insects produced shorter periods of Pattern 4 on moderately resistant IR46 and resistant IR62 rice varieties than on susceptible IR22, although the time taken to produce the first period of Pattern 4 from the start of EPG recording did not differ between varieties. This suggests that N. lugens has the ability to initiate phloem ingestion on all three varieties, but that resistance mechanisms in IR46 and IR62 prevent sustained phloem ingestion, at least within the first 8 h of contact.
Résumé Le comportement alimentaire de N. lugens Stahl a été enregistré életroniquement pendant 8 heures aux Philippines sur 3 variétés de riz présentant différents degrés de résistance, en utilisant un amplificateur DC associé à un système vidéo pour enregistrer simultanément la production de miellat.Six types d'enregistrements de pénétration (EPG) sont décrits avec leurs caractéristiques de fréquence et d'amplitude. Les types 4 et 5 sont associés à la production de 2 types différents de miellat; le miellat de base contenant des acides aminés est produit pendant le type 4, et le miellat acide sans acides aminés est produit pendant le type 5. On considére que ces types indiquent respectivement l'ingestion de phloème et xylème par N. lugens.Les périodes de type 4 ont été plus courtes sur les variétés modérément résistante IR46 et résistante IR62 que sur la variété sensible IR22, bien que le temps passé à produire la première période de type 4 depuis le début des enregistrements des EPG ne diffère pas suivant les variétés. Ceci laisse penser que N. lugens a la possibilité de commencer la consommation de phloème sur les trois variétés, mais que les mécanismes de résistance chez IR46 et IR62 empêchent une ingestion soutenue de phloème, au moins pendant les 8 premières heures de contact.
  相似文献   

19.
Oviposition behavior of Nilaparvata lugens (Stål) (Homoptera: Delphacidae) on rice, Oryzae sativa L., was monitored using a videocamera and an AC (alternating-current) electronic measuring system. Oviposition by N. lugens always took place during stylet penetration into the plant. After penetration of the ovipositor, there were three specific ovipositional sequences observed; sawing insertion of the ovipositor, release of an egg in the plant, and partial or complete withdrawal of the ovipositor from the plant. Distinctive waveforms were recorded in correlating with these events. From a sequence of waveform patterns, it was possible to determine the number of eggs and egg masses laid inside the plant tissue and the duration of oviposition behavior. Waveform analysis revealed that N. lugens laid an egg mass consisting of on average 5.7 eggs. This occurred 4.3 min after a period of about 6.4 min of stylet penetration, and this sequence was repeated several times in 24 h. Electrical and/or tethering effects on oviposition appear to be negligible within 24 h of recording.  相似文献   

20.
The effect of expressing the gene encoding snowdrop lectin (Galanthus nivalis agglutinin, GNA) in transgenic potato plants, on parasitism of the phytophagous insect pest Lacanobia oleracea by the gregarious ectoparasitoid Eulophus pennicornis, was investigated in glasshouse trials. Expression of GNA (approx. 1.0% total soluble protein) by transgenic plants significantly reduced the level of pest damage, thus confirming previous studies. Furthermore, the presence of the parasitoid significantly reduced the levels of damage incurred either by the transgenic or control plants when compared to those plants grown in the absence of the parasitoid. For the GNA expressing plants the presence of the parasitoid resulted in further reductions (ca. 21%) in the level of damage caused by the pest species. The ability of the wasp to parasitise and subsequently develop on the pest larvae was not altered by the presence of GNA in the diet of the host. E. pennicornis progeny that developed on L. oleracea reared on GNA expressing plants showed no significant alteration in fecundity when compared with wasps that had developed on hosts fed on control potato plants, although mean size and longevity of female parasitoids was significantly reduced. The number of F 2 progeny produced by parasitoids derived from hosts fed on GNA-expressing plants was not significantly different to those produced by parasitoids from hosts fed control plants. Results from the present study demonstrate that the use of transgenic plants expressing insecticidal proteins can be compatible with the deployment of beneficial insects and that the two factors may interact in a positive manner.  相似文献   

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