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1.
马铃薯甲虫是重要的入侵害虫,严重威胁着我国粮食作物马铃薯的生产。苏云金芽胞杆菌是重要的农业害虫生防细菌,对马铃薯甲虫有良好的防治效果。本文围绕苏云金芽胞杆菌在马铃薯甲虫防治上的研究进展与应用进行综述。主要从马铃薯甲虫的入侵与防治手段、苏云金芽胞杆菌的晶体蛋白结构与杀虫机制、对马铃薯甲虫有活性的Bt毒蛋白研究进展、Bt毒蛋白对马铃薯甲虫的作用机制以及马铃薯甲虫对Bt毒蛋白的抗性机制等方面进行了综述。最后,从Bt新基因的挖掘和杀虫机理方面对苏云金芽胞杆菌在马铃薯甲虫防治上的研究进行了展望。  相似文献   

2.
BACKGROUND: Ceratitis capitata (Wiedemann) is one of the world's most destructive fruit pests. The aim of this study was to ascertain insecticidal activity of Bacillus thuringiensis (Berliner) δ‐endotoxins to C. capitata. RESULTS: Among 42 selected Bacillus strains, only B. thuringiensis serovar israelensis (Bti) solubilised protoxins showed biological activity against C. capitata neonate larvae, whereas Bti spore and crystal mixture was inactive. Insecticidal activity of Bti protoxins was significantly enhanced by incubation with Culex pipiens L. gut extracts. Overdigestion of Bti protoxins with Sesamia nonagrioides (Lefebvre) gut extracts suppressed biological activity against C. capitata, and this correlated with degradation of Cyt toxins. Cyt1Aa solubilised protoxin showed the highest toxicity, LC50 after 7 days of 4.93 µg cm?2, while proteolytical processing of Cyt1Aa protoxins by larval gut extracts did not enhance insecticidal activity. CONCLUSION: The present study provides evidence for the first time of the insecticidal activity of a B. thuringiensis strain against C. capitata and identifies a single δ‐endotoxin with potential for controlling this pest. Copyright © 2010 Society of Chemical Industry  相似文献   

3.
为分离并鉴定对亚洲玉米螟具有高毒力的苏云金芽胞杆菌菌种,通过棋盘式采集法从吉林省农业科学院试验田土壤中分离获得野生菌株,进行了形态、生化特性、伴孢晶体观察及基因型鉴定,并测定了其对3种鳞翅目害虫的室内生物活性。结果显示:该菌株在LB培养基上为圆形、暗白色菌落,边缘不整齐,革兰氏染色阳性,产生的伴孢晶体形状多为不规则形。16S r DNA序列与苏云金芽胞杆菌属的NBRC 13865同源性达99%;该菌株为cry2+cry9复合基因型,编码氨基酸序列与Cry2Ab和Cry9Ea蛋白同源性分别为94%和99%,鉴定为苏云金芽胞杆菌,命名为JN001。该菌株对亚洲玉米螟具有较高毒性,以1.0×109个/m L菌体浓度接种3龄幼虫72 h后,其校正死亡率为95.06%,致死中浓度(LC50)为4.12×103个/m L菌体,而对斜纹夜蛾和粘虫毒性较弱,接种72 h后校正死亡率分别为12.88%和7.34%。表明该菌株是1株对亚洲玉米螟具有较强毒力的苏云金芽胞杆菌新菌株,具有较好的开发利用潜力。  相似文献   

4.
Insecticidal δ‐endotoxin proteins, degraded from parasporal crystals by protease, were recovered by a simple procedure using heat treatment, solubilization, and ultrafiltration of a fermentation broth of Bacillus thuringiensis subsp kurstaki HD‐1. A 68 kDa insecticidal protein was obtained and characterized by SDS‐PAGE. The procedure described gave a nearly quantitative recovery of toxicity. Furthermore, bioassay results on larvae of the diamondback moth (Plutella xylostella) showed that the 68 kDa δ‐endotoxin fraction (P1) was the principal insecticidal component to this target insect. A similar molecular mass polypeptide P2 (65 kDa) which was solubilized together with P1 from the parasporal crystals, gave relatively low mortality. © 2000 Society of Chemical Industry  相似文献   

5.
采用桑叶浸渍法和饲料表面覆盖法测定发现,从土壤中分离的一株苏云金芽胞杆菌菌株GS8对家蚕Bombyx mori、甜菜夜蛾Spodoptera exigua和棉铃虫Helicoverpa armigera初孵幼虫均表现出较高的杀虫活性,经形态学和生理生化反应鉴定该菌株为东北亚种(Bacillus thuringiensis subsp.tohokuensis)。聚丙烯酰胺凝胶电泳(SDS-PAGE)分析结果表明,GS8菌株的杀虫蛋白晶体主要由分子质量为130、81和60 kDa的蛋白组成。聚合酶链反应-限制性片段长度多态性(PCR-RFLP)鉴定结果显示,GS8菌株含有 cry1Aa、cry1Ab、cry1Ib、cry2Ab和cry9Ba 等基因。  相似文献   

6.
[目的]从北京植物园采集的土壤样品中分离高毒力的苏云金芽胞杆菌.[方法]采用温度法筛选Bt菌株,比对大质粒DNA图谱,区分不同类型的Bt菌株,PCR-RFLP方法对cry1~cry40类基因型进行鉴定,SDS-PAGE分析杀虫晶体蛋白,测定Bt分离株对大猿叶甲、小菜蛾幼虫的杀虫活性,筛选出高毒力的菌株.[结果]从149份土壤样品中分离出147株Bt,分为12种类型,含有菱形、方形、球形和不规则等晶体类型;6株菌中分别含有fry1Aa、cry1Ab、cry1Ac、cy1Ah、cry1Ba、cry1Be、cry1Ia、cry1La、cry2Ab和cry7Aa等基因类型,其余6株中不含有已知基因.有9株表达约130 ku蛋白,1株表达约70 ku蛋白,2株表达约150 ku蛋白,有3株既表达130 ku蛋白又表达60 ku蛋白;发现一株对大猿叶甲具有较高毒力的菌株ZWY-7,1株对小菜蛾有高毒力的菌株ZWY-9,2株菌都没有检测到已知基因型.[结论]北京植物园中Bt分布广泛,类型多样,其中发现两株高毒力的菌株,有望获得新的杀虫基因.  相似文献   

7.
BACKGROUND: Supernatant‐secreted proteins in Bacillus thuringiensis (Berliner) (Bt) with insecticidal activity provide an important source of information for discovery of new useful strains and/or entomotoxins. However, physiological variation among isolates might interfere in the detection efficiency of screening procedures on Bt collections. The aim of this study was to assess the magnitude of this variation in a sample of isolates from a tropical Bt collection, which was gauged through the assessment of their temporal patterns of growth and protein secretion in culture supernatants (SNs), as well as of the corresponding toxicity against fall armyworm (Spodoptera frugiperda, JE Smith). Feeding bioassays were performed, with larvae being treated with heated and non‐heated total protein extracted from SNs collected at different culture times. Larva mortality and reduction in pupa formation were observed. RESULTS: Intra‐ and interisolate variations were observed in the temporal patterns of growth, quality and quantity of protein secreted, as well as in insecticidal activity of these SNs, based on larvae mortality and pupation rates. These results suggest that the insecticidal potential of certain isolates can be hidden if comparisons are done on the basis of the same number of cells in the culture and/or the same culturing time. CONCLUSIONS: Methods of screening Bt collections on the basis of feeding bioassays can be misleading with regards to identifying more promising isolates for biocontrol purposes if physiological differences are not considered. The consequences and implications of these findings for the development of experimental systems that depend on toxicity bioassays to identify alternative Bt strains and entomotoxins with practical applicability have been discussed. Copyright © 2011 Society of Chemical Industry  相似文献   

8.
BACKGROUND: Biopesticides containing Cry insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are effective against many lepidopteran pests, but there is a lack of Bt‐based pesticides for efficient control of important coleopteran pests. Based on the reported increase in Bt toxin oligomerization by a polypeptide from the Cry3Aa receptor cadherin in Tenebrio molitor (Coleoptera: Tenebrionidae), it was hypothesized that this cadherin peptide, rTmCad1p, would enhance Cry3Aa toxicity towards coleopteran larvae. To test this hypothesis, the relative toxicity of Cry3Aa, with or without rTmCad1p, against damaging chrysomelid vegetable pests of China was evaluated. RESULTS: Cry3Aa toxicity was evaluated in the spotted asparagus beetle (Crioceris quatuordecimpunctata), cabbage leaf beetle (Colaphellus bowringi) and daikon leaf beetle (Phaedon brassicae). To assess the effect of rTmCad1p on Cry3Aa toxicity, neonate larvae were fed Cry3Aa toxin alone or in combination with increasing amounts of rTmCad1p. The data demonstrated that Cry3Aa toxicity was significantly increased in all three vegetable pests, resulting in as much as a 15.3‐fold increase in larval mortality. CONCLUSION: The application of rTmCad1p to enhance Cry3Aa insecticidal activity has potential for use in increasing range and activity levels against coleopteran pests displaying low susceptibility to Bt‐based biopesticides. Copyright © 2011 Society of Chemical Industry  相似文献   

9.
自青海北山林场桦树根围分离筛选到芽胞杆菌菌株166株,根据菌落特征分为6个类群,选择其中31株菌株纯化保存并进行BOX—PCR及ERIC—PCR指纹图谱分析、16SrDNA及gyrB基因序列分析,鉴定出球形赖氨酸芽胞杆菌Lysinibacillussphaericus12株、解淀粉芽胞杆菌Bacillusamyloliq-uefaciens 5株、韦氏芽胞杆菌Bacillusweihenstephanensis7株、芽胞杆菌Bacillussp.5株、苏云金芽胞杆菌Bacillusthuringiensis2株。通过平板对峙试验筛选得到11株对油菜菌核病原真菌Sclerotiniasclerotiorum具有显著拮抗活性的菌株。以MALDI.TOF.MS质谱分析拮抗菌株QBll的活性成分,可产生脂肽化合物表面活性素和泛革素,表明其拮抗活性与脂肽化合物的合成和分泌有关。  相似文献   

10.
对基因工程菌Escherichia.coli fss6所产黑色素进行了紫外保护研究。结果表明:添加黑色素的苏云金芽孢杆菌(Bt)经紫外照射后菌体的存活率是不加黑色素菌体的8倍;黑色素能有效保护Bt的晶体毒素蛋白免受紫外照射的降解;随着被照射菌体中黑色素浓度的增加,其杀虫活性也相应提高。  相似文献   

11.
The extensive mammalian toxicity studies performed to support the safety of Bacillus thuringiensis-containing pesticides clearly demonstrate that the tested isolates are not toxic or pathogenic. Toxicity studies submitted to the US Environmental Protection Agency to support the registration of B. thuringiensis subspecies have failed to show any significant adverse effects in body weight gain, clinical observations, or upon necropsy. Infectivity/pathogenicity studies have shown that the intact rodent immune system responds as expected to eliminate B. thuringiensis gradually from the body after oral, pulmonary or intravenous challenge. Similar clearance patterns were observed with B. subtilis and B. sphaericus, which also were non-toxic to test animals. The results also indicate that the currently used protocols for toxicity/pathogenicity evaluations of micro-organisms in laboratory animals have provided useful and necessary information for risk assessment. The study results raise the issue, however, of whether some reduced set of toxicily/pathogenicity studies could be required for new B. thuringiensis isolates of already registered subspecies, and whether this reduced data set would also apply to subspecies not yet registered as pesticidal active ingredients.  相似文献   

12.
为获得对大黑鳃金龟甲幼虫具有较高杀虫活性的菌株,利用拌土法测定了本实验室分离的500株苏云金芽胞杆菌(Bacillus thuringiensis,Bt)对大黑鳃金龟甲的杀虫活性,建立Bt菌株比较鉴别技术来进行多样性分析,并对菌株的晶体形态及幼虫感染Bt后的中肠组织切片进行观察。结果显示,从500株Bt菌株中筛选到42株对大黑鳃金龟甲幼虫具有不同程度活性的菌株,分属于14个菌株类型;其中有10株校正死亡率大于60%,261-1菌株杀虫活性最高,7 d校正死亡率达100%。从14个菌株类型中各选取1个代表菌株进行基因鉴定,仅P65-1、1126-1、FCD114和78-2菌株分别含有cry8Ma、cry8Ca、cry8Ab、cry8Ga、cry8Ea共5个cry8类新基因,其它10个菌株均不含cry3、cry8、cry18、cry23、cry37、cry43等对鞘翅目害虫有效的杀虫基因。14个菌株中,78-3、127-2和1198-1菌株能产生双锥体型晶体,261-1、FCD114、P65-1、FTL84、78-2和1126-1菌株能产生球形晶体。幼虫感染261-1、1198-1、FCD114、1126-1、QDL40-2菌株后,中肠组织发生明显的病理变化:2 d时肠壁细胞明显出现空洞化,排列疏松,4 d时受到严重破坏并脱离底膜。表明筛选到的Bt菌株具有防治大黑鳃金龟甲幼虫的潜力。  相似文献   

13.
14.
Bacillus thuringiensis cotton is a variety of cotton genetically modified to contain a gene derived from B. thuringiensis bacteria; which results in expression of toxin protein that confers resistance to bollworm complex (the most destructive pest of cotton). Introduction of Bt cotton lowered the need of insecticides, but still a number of insecticides are used for other insects like jassids, whitefly, aphids and tobacco caterpillar to which Bt gene does not provide effective control. Imidacloprid (tradename Imidacel 17.8 SL) is an insecticide designed for control of these major sucking/piercing insects that affect cotton. So in the present work we studied the post effect of imidacloprid insecticide on plant health of three Bt cotton hybrids (RCH-134, JKCH-1947, NCEH-6R) as there are reports of this insecticide causing growth and yield enhancements in absence of insect pests. Imidacloprid was first sprayed at recommended concentration (40 ml/acre) on 3 months old plants sown in randomly designed plots with three replications of each hybrid. The spray was repeated three times at 10 days interval. The level of B. thuringiensis gene expression, peroxidase activity and total phenols was measured on third day after every spray in leaves along with growth and yield of plants. The insecticide has shown to increase the level of B. thuringiensis protein, peroxidase enzyme activity, total phenols, height, number of bolls retained on plants and yield. These observations suggested that the imidacloprid treated plants showed better growth and development, thereby imidacloprid has growth enhancing effect on Bt cotton plants in addition to its insecticidal properties.  相似文献   

15.
从钙粘蛋白介导苏云金杆菌晶体(Bt Cry)毒素对害虫的毒杀过程、钙粘蛋白重复区和近膜区与Bt Cry毒素的分子间作用涉及的结合位点及可能的互作机制等方面,综述了钙粘蛋白片段与Bt Cry毒素协同作用的最新研究进展。昆虫钙粘蛋白某些片段在非折叠状态时,可与Bt Cry毒素形成寡聚体,从而增加Bt Cry毒素对靶标害虫的毒杀活性。相关研究成果有助于提高Bt Cry毒素毒杀害虫的能力,克服害虫抗药性,具有一定的应用前景。  相似文献   

16.
BACKGROUND: Diamondback moth, Plutella xylostella (L.), is a major insect pest of crucifers in the biodiversity‐rich north‐western Indian Himalayan hills. The present investigation was aimed at determining the susceptibility pattern of P. xylostella populations collected from different locations of this region to autochthonous and standard Bacillus thuringiensis strains. RESULTS: Among the reference as well as indigenous B. thuringiensis strains tested, sub spp. kurstaki HD‐1, kurstaki HD‐73, galleriae HD‐8, local galleriae/colmeri strain BtOa1 and some of their Cry1 class toxins were found to be highly toxic. Surprisingly, the sub sp. tolworthi HD‐125, local tolworthi strain BtHa1 and Cry9 class toxins were found to be non‐toxic. Midgut homogenate from fourth‐instar larvae was found to activate 130 kDa protoxin from the local tolworthi strain BtHa1 into 68 kDa toxin, but failed to exert any larval mortality, probably owing to lack of receptor binding. CONCLUSION: The present study provides valuable baseline susceptibility data for the deployment of B. thuringiensis‐based control methods, as well as for future monitoring of development of resistance in P. xylostella to B. thuringiensis in this ecologically sensitive region. Copyright © 2008 Society of Chemical Industry  相似文献   

17.
A local isolate of Metarhizium anisopliae (Hypocreales: Clavicipitaceae), Bacillus thuringiensis subsp. kurstaki and chlorantraniliprole were assessed against six field populations of tomato fruitworm Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in a series of laboratory bioassays. Two dose rates of B. thuringiensis (0.5, 1 μg g?1), one of both M. anisopliae (1.3?×?106 conidia ml?1) and chlorantraniliprole (0.01 ppm) were applied alone and in combination with each other against 2nd, 3rd, 4th and 5th larval instars. The mortality was observed every 24 h until pupation. The bioassays were carried out at 25°C and 75% r.h. The highest mortality was observed in Rawalpindi with the lowest pupation rate by applying the combined concentrations of B. thuringiensis and chlorantraniliprole. The lowest mortality was observed in population from Gujranwala among all the tested populations. The antagonistic interaction was noted where the high dose rate of B. thuringiensis was combined with M. anisopliae; however, the remaining interactions enhanced the mortality and reduced the percent pupation. The overall results demonstrated that all the treatments gave significant control of the larval instars of H. armigera. The population from Gujranwala proved least susceptible whereas the one from Rawalpindi was highly susceptible.  相似文献   

18.
In spring 1972 experiments were carried out in Lower Austria with the Bacillus thuringiensis isolate Dipel in order to assess its effectiveness in controlling Colotois pennaria L. infesting oak and other broad-leaved trees. The results obtained compared favourably with those produced by the insecticides Nexion2), malathion and Dipterex3), although both malathion and Dipterex yielded higher mortality. Similar results were obtained when B. thuringiensis was used to control Stilpnotia salicis L., feeding on poplar trees. Neodiprion sertifer Geoffr. and Pris-tiphora abietina Christ., however, were not affected.  相似文献   

19.
从自然死亡的玉米螟幼虫体内分离获得一株对玉米螟高毒的苏云金芽孢杆菌,编号为HB-3。该菌株产生菱形和方形两种晶体蛋白。纯化晶体及孢晶混合物的毒力测定结果表明,该菌株对玉米螟具有较高毒力,LC50分别为5.82μg/mL和5.94μg/mL,对尖音库蚊也有一定毒力,LC50分别为16.59μg/mL和17.50μg/mL。生理生化特性研究结果表明,HB-3与已知的蜡螟亚种(Bacillus thuringiensissubsp.galleriae)基本一致,血清型为H5ab。晶体蛋白的SDS-PAGE分析表明,HB-3菌株晶体蛋白至少由分子量为133 kDa、65 kDa4、5 kDa和23 kDa的4种多肽组成。  相似文献   

20.
The European corn borer (Ostrinia nubilalis), the Mediterranean corn borer (Sesamia nonagrioides) and the western corn rootworm (Diabrotica virgifera virgifera) are the main arthropod pests in European maize production. Practised pest control includes chemical control, biological control and cultural control such as ploughing and crop rotation. A pest control option that is available since 1996 is maize varieties that are genetically engineered (GE) to produce insecticidal compounds. GE maize varieties available today express one or several genes from Bacillus thuringiensis (Bt) that target corn borers or corn rootworms. Incentives to growing Bt maize are simplified farm operations, high pest control efficiency, improved grain quality and ecological benefits. Limitations include the risk of resistance evolution in target pest populations, risk of secondary pest outbreaks and increased administration to comply with licence agreements. Growers willing to plant Bt maize in the European Union (EU) often face the problem that authorisation is denied. Only one Bt maize transformation event (MON810) is currently authorised for commercial cultivation, and some national authorities have banned cultivation. Spain is the only EU member state where Bt maize adoption levels are currently delivering farm income gains near full potential levels. In an integrated pest management (IPM) context, Bt maize can be regarded as a preventive (host plant resistance) or a responsive pest control measure. In any case, Bt maize is a highly specific tool that efficiently controls the main pests and allows combination with other preventive or responsive measures to solve other agricultural problems including those with secondary pests. Copyright © 2011 Society of Chemical Industry  相似文献   

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