Susceptibility pattern and development of resistance in the diamondback moth,Plutella xylostella L,to Bacillus thuringiensis Berl var kurstaki in India

Base‐line susceptibility for six‐day‐old larvae of the diamondback moth, Plutella xylostella, against Bacillus thuringiensis var kurstaki (Biobit^®) was studied by a cabbage leaf disc dip bioassay technique. Diamondback moth from 13 locations in seven different states spread over a distance of about 3000 km longitudinally was used for these studies. Forty‐eight‐hour LC₅₀ values varied from 1.0 to 10.97 mg AI litre⁻¹. Further investigations on the development of resistance under laboratory conditions showed an increase in LC₅₀ from 2.76 (for unselected F₁ generation) to 5.28 mg AI litre⁻¹ (for selected F₉ generation), using a selection concentration of 6.4 mg AI litre⁻¹. This suggested a possibility of the development of resistance under field conditions if there were to be extensive and indiscriminate use of B thuringiensis. These findings are discussed in relation to integrated pest management and the mechanisms of resistance in resistance management tactics. © 2000 Society of Chemical Industry     

Bt制剂防治玉米螟的田间效果

研究了Bt、克百威、丁硫克百威、溴氰菊酯、杀虫双和辛硫磷6种药剂处理对玉米螟的田间防治效果。结果表明,6种药剂均能显著降低玉米的被害株率,克百威的防效最好,其次为丁硫克百威、辛硫磷和溴氰菊酯,对被害株的相对防效分别为71.70%、70.03%、68.47%和68.11%,Bt生物制剂的防效达66%以上;6种药剂对幼虫的相对防效均达到80%以上;Bt生物制剂的防效与其他化学制剂间差异不显著（p<0.01）。辛硫磷处理易发生药害,但5 d后叶片白斑逐渐消失。     

具杀线虫活性的苏云金杆菌筛选研究

 Bioassay of 51 strains of Bacillus thuringiensis to Bursaphelenchus xylophilus had been studied in laboratory. Two of them had higher toxicity to B. xylophilus. Extraction of endotoxin and exotoxin of the two isolates to B. xylophilus showed the action of exotoxin higher than that of endotoxin.     

吐温80对Bt在作物叶面的展着及葱田甜菜夜蛾防效的影响

在苏云金杆菌中,加入表面活性剂吐温80,研究了苏云金杆菌在不同作物上的展着及对葱田甜菜夜蛾防效的影响。试验结果表明,吐温80能显著提高Bt在作物叶面的展着量。药液在不同作物的展着量差异显著,吐温80的2 000倍稀释液,可使CAB109制剂在葱、玉米、羽衣甘蓝、菜豆、草莓、白菜、黄瓜和萝卜的展着量分别提高8.0、7.9、7.7、2.5、2.3、2.0、1.4倍和1.3倍;可使商品TB的展着量分别提高4.4、6.4、4.7、1.8、1.9、1.7、1.4倍和1.2倍。吐温80的1 000倍稀释液,可使CAB109与TB制剂将葱田甜菜夜蛾的虫口减退率分别从63.6%与43.3%,提高到84.5%与62.8%,显著提高防效。     

Effects of interactions between host plants and selective insecticides on larvae of Plutella xylostella L. (Lepidoptera: Yponomeutidae) in the laboratory

The residual toxicity of two selective insecticides, teflubenzuron (acylurea) and Bacillus thuringiensis Berliner ssp. aizawai (microbial), to laboratory and field strains of Plutella xylostella L. was shown in the laboratory to be significantly affected by leaf nutritional status, other host-plant resistance factors, cultivation method and plant age. With plants offering some degree of host-plant resistance, the toxicity of the insecticides was either increased or decreased compared with highly susceptible plants, depending on the specific nature of the plant-herbivore interaction. Differences in residual toxicity of the insecticides varied up to nine-fold on different host plants (=host-plant- + insecticide-induced mortality) despite less than four-fold differences in P. xylostella mortality in controls (=host-plant-induced mortality alone). Host-plant nutritional status also had a substantial effect on the damage potential of larvae. Different response times by P. xylostella to the two insecticides tested on host plants of varying nutritional status were related to the contrasting modes of action of the respective active ingredients. The present studies suggest that insecticides applied to Brassica oleracea L. var. capitata with partial plant resistance may contribute to improved control of P. xylostella. A conceptual model is used to describe likely mortality responses by macrophagous larvae to insecticides applied to plants of varying resistance status. The implications of the findings in relation to the integrated management of P. xylostella are considered.     

云南小菜蛾对Bt的抗药性及盆栽药效评价

2008-2012年, 采用浸叶法连续5年测定了云南省2个主要蔬菜种植区通海县和弥渡县的小菜蛾田间种群对Bt制剂的抗药性,并进行了盆栽药效评价。结果表明,两地区小菜蛾对Bt制剂均表现为低抗或无抗性,抗药性无明显上升趋势。比较了在光照培养箱和自然条件下3种不同Bt制剂对通海和弥渡县小菜蛾盆栽药效,药后第5天通海和弥渡小菜蛾种群在恒温（25±1 ℃,RH65%~70%）和自然条件下(12~28 ℃,RH50%~75%)的校正死亡率分别为88.3%~95.5%和80.2%~90.6%、78.8%~89.1%和73.4%~85.7%,两者差异不显著。云南小菜蛾种群对Bt制剂均有较高的敏感性,可在十字花科蔬菜生产中有选择地使用。     

Differential toxicity of Bacillus thuringiensis strains and their crystal toxins against high-altitude Himalayan populations of diamondback moth, Plutella xylostella L

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     

Activity of Bacillus thuringiensis toxins against cocoa pod borer larvae

Twelve Cry proteins from Bacillus thuringiensis Berliner were tested in bioassays on cacao plantations in Indonesia for activity against the larvae of cocoa pod borer (Conopomorpha cramerella (Snellen)), an insect pest of the cacao tree. Through the damage caused by their feeding, the larvae of cocoa pod borer cause the pods of the cocoa tree to ripen prematurely. They are difficult to control with conventional measures. Preliminary assays identified five toxins that were more active than others. In two subsequent bioassays the activity of selected toxins was determined more accurately. Three Cryl proteins with relatively little homology were all found to be toxic, opening perspectives for controlling cocoa pod borer by expression of Cry proteins in transgenic plants.     

四株对小地老虎有活性的苏云金芽胞杆菌的毒力评价

本文对4株苏云金芽胞杆菌Bacillus thuringiensis（Bt）的杀虫基因类型、杀虫蛋白表达类型、蛋白表达量以及杀虫活性进行了初步的评价分析。菌株PS9-D12和PS9-C12的基因类型与蛋白表达类型丰富,且其杀虫蛋白表达量是对照菌株HD-1的1.7倍。在相同培养条件下,制备菌株胞晶混合冻干粉对小地老虎Agrotis ypsilon的生测结果显示,菌株HD-1、PS9-D12、PS9-C12、PS9-D11和PS9-H9的LC50分别为0.71、0.19、0.14、0.24和1.16 mg·g^-1,菌株PS9-D12和PS9-C12的杀虫活性显著高于菌株HD-1（P〈0.05）;此4株菌株对小菜蛾Plutella xylostella幼虫的校正死亡率均大于78%,它们对甜菜夜蛾Spodoptera exigua幼虫的活性均高于菌株HD-1;而对大猿叶甲Colaphellus bowringi幼虫均无活性。可见菌株PS9-D12和PS9-C12比商业化生产的HD-1有更好的杀虫活性,可作为Bt杀虫剂的候选材料。     

研究苏云金芽孢杆菌毒素作用的新思路

苏云金芽孢杆菌对多种昆虫具有特异毒杀作用,虽然对其作用机理有了一定的了解,但对昆虫随之出现的抗性研究还很不深入。利用秀丽隐杆线虫这种模式动物并结合基因芯片技术和基因组学研究,为详细了解苏云金芽孢杆菌毒素的作用机理、昆虫对毒素的抗性机理和研究更加高效的生物杀虫制剂提供了新的思路。     

苏云金杆菌小试生产发酵影响因子的研究

利用 1 0L全自动发酵罐对苏云金杆菌小试生产的发酵条件进行了研究。结果显示 ,不同的温度、pH值、培养基成分及接种方式对发酵产量和发酵周期均有影响 ;在培养基配方中存在葡萄糖时 ,葡萄糖单独灭菌对提高苏云金杆菌发酵产量是重要的。初步生物测定显示 ,发酵上清中存在一些杀虫增效因子 ,建议配制苏云金杆菌制剂时尽可能加以利用。     

苏云金芽胞杆菌安全性的研究进展

苏云金芽胞杆菌（Bacillus thuringiensis,简称Bt）是目前应用最为广泛的一类微生物农药。本文分析比较了我国及美国Bt产品登记和安全评价等情况,并对Bt制剂的风险评价进行了分析。Bt菌株不是人或其他哺乳动物的已知病原体,对哺乳动物无毒性、感染性和致病性;其种群数量在环境中呈逐渐减少趋势,无残留风险;对非靶标昆虫的生存和繁殖及土壤微生物群落结构无显著影响。     

Inheritance of resistance to a Bacillus thuringiensis toxin in a field population of diamondback moth (Plutella xylostella)

Inheritance of resistance to the Bacillus thuringiensis Berl. CryIA(b) crystal protein was studied in Plutella xylostella L. (diamondback moth). A field population 50-fold more resistant to CryIA(b) than a control susceptible strain was used. Dose-mortality curves of the resistant population, the susceptible strain and the F₁ from the two reciprocal crosses were compared. Resistance transmission to the F₁ was dependent on the sex of the resistant progenitor. Sex ratio of the survivors to high doses of CryIA(b) in the F₁ of the two reciprocal crosses did not corroborate the preliminary hypothesis of resistance being due to a recessive sex-linked allele. Since, in a previous work, the loss of CryIA(b) binding capacity of resistant insects had been demonstrated, binding to midgut tissue sections from F₁ individuals was also analysed. The presence of binding in all of the F₁ preparations showed that, at least, a recessive autosomal allele was responsible for the loss of binding capacity in the resistant population.     

苏云金芽胞杆菌JQ23的表型鉴定及对韭菜迟眼蕈蚊的防治效果

韭菜迟眼蕈蚊是韭菜生产上的重要害虫,实验室从河南省采集的土样中分离获得1株对韭菜迟眼蕈蚊幼虫高毒力的Bt新菌株JQ23。经BiologGENⅢ细菌鉴定系统分析菌株JQ23为苏云金芽胞杆菌,相似率为0.596;通过室内生物活性测定,明确菌株JQ23对韭菜迟眼蕈蚊2龄幼虫72h的LC₅₀值为8.38×10⁶芽胞/mL;经室内盆栽防治试验结果表明,菌株JQ23在1×10⁷芽胞/mL浓度下,采用滴灌法连续用药3次,间隔7d,对韭菜迟眼蕈蚊的防治效果达到86.28%;经春季田间小区试验,再次验证菌株JQ23在1×10⁸芽胞/mL浓度下,采用滴灌法连续用药3次,间隔7~10d,对韭菜迟眼蕈蚊的防治效果达到66.75%。由此可见,菌株JQ23能够有效地控制韭菜迟眼蕈蚊的虫口数量,减少作物的受害率,达到较好的防治效果。     

对植物寄生线虫具有活性的苏云金芽胞杆菌研究进展

植物寄生线虫是重要的植物病原物之一,给农业生产带来了巨大的经济损失。长期以来,一直缺乏有效防治植物寄生线虫的手段。苏云金芽胞杆菌（Bacillus thuringiensis,Bt）是重要的昆虫病原细菌,广泛应用于鳞翅目、双翅目、鞘翅目害虫等农林及卫生害虫的防治;部分Bt菌株对植物寄生线虫具有很高的活性。本文总结了杀植物寄生线虫Bt菌株筛选的模型与方法的建立、Bt菌株杀植物寄生线虫的作用机理及其相关应用、以及杀植物寄生线虫的伴胞晶体蛋白与相关基因等,可望为杀植物寄生线虫Bt制剂的研究与开发提供借鉴。     

苏云金芽胞杆菌杀线虫活性因子分析及杀虫机制研究进展

苏云金芽胞杆菌Bacillus thuringiensis(Bt),是一种重要的昆虫病原细菌,对多种农业害虫包括鳞翅目、鞘翅目、双翅目、膜翅目和线虫等具有高特异杀虫活性,是迄今应用最广泛的微生物杀虫剂。本文总结了杀线虫Bt菌株的活性因子种类以及不同类的活性物质杀线虫机制的分析,能够为生物杀线剂的研究与创制提供理论基础。     

具杀松材线虫活性的苏云金芽胞杆菌筛选及其毒力测定

松材线虫是松树萎蔫病的主要病原,为了筛选到具有较高杀线虫活性的苏云金芽胞杆菌Bacillus thuringiensis菌株,采用直接触杀法对苏云金芽胞杆菌发酵液进行杀线虫活性以及稳定性的检测,并对筛选出的高活性菌株通过扩增16S rDNA片段进行菌株的鉴定。结果表明,4个菌株中具有最高活性的菌株D发酵液处理松材线虫8 h的死亡率超过70%,24 h其死亡率达到100%;在温度为-20、25、40、60、80和121℃条件下处理的菌株D发酵液,其对松材线虫的毒力都在85%以上;在pH 5.0~10.0时,菌株D发酵液对松材线虫的毒力都在85%~94%,并无显著的差异;将菌株D发酵液进行蛋白酶K处理后,其对松材线虫的毒力仍在98%以上;在4℃环境下保存50 d,菌株D发酵液的毒力没有明显下降。通过16S rDNA 1.5 kb片段的序列分析,初步得出菌株D可能是苏云金芽胞杆菌KNU-07。可见,菌株D发酵液具有耐热、耐储藏和对蛋白酶K稳定的特性,是一株具有开发潜力和应用前景的高毒力菌株。     

重组增效蛋白对Bt和氯氰菊酯防治棉铃虫的增效作用

前期在大肠杆菌中表达得到具有显著生物活性的含粉纹夜蛾颗粒体病毒增效蛋白C端818个氨基酸的融合蛋白P96。以毒饲料法研究P96对苏云金芽孢杆菌和氯氰菊酯杀棉铃虫敏感、抗性品系的增效作用。结果表明：P96对Bt杀棉铃虫敏感、低抗、中抗品系的增效比分别为3．34、4．72和9．82；对氯氰菊酯杀棉铃虫敏感、低抗、中抗品系的增效比分别为2．53、3．38和6．24。随棉铃虫抗性升高，P96的增效作用愈为明显。单对汰选是快速选育棉铃虫抗Bt品系的有效方法。     

基于比较基因组学分析苏云金芽胞杆菌防治蛴螬的相关功能基因

为挖掘对地下害虫蛴螬高效的苏云金芽胞杆菌（Bacillus thuringiensis,简称Bt）菌株在杀虫及环境适应方面的关键功能基因,本研究对蛴螬高效的菌株261-1、Fcd114、1126-1、HBF-18和78-2以及对鳞翅目害虫高效菌株WBt-2、G03进行了基因组测序,并与对鳞翅目害虫高效菌株HD1、HD12、HD73和对蛴螬高毒力菌株Bt185等基因组进行了比较分析,重点分析比较了杀虫基因、多糖代谢相关基因、与其他微生物竞争和适应性相关的酰基高丝氨酸内酯水解酶（N-Acylhomoserine lactones hydrolase,aiiA）、突破和克服靶标昆虫免疫系统的免疫抑制因子A（Immune Inhibitor A,InhA）、具有抗病并能增效杀虫的双效菌素（Zwittermicin A,ZwA）等五大类基因和基因簇。研究发现,这些菌株按照其所含有的杀虫基因特异性可分为3个类型：对鳞翅目害虫有效的菌株（类型1）、对鞘翅目害虫有效的菌株（类型2）以及同时含有两种害虫有效基因的菌株（类型3）。对地下害虫蛴螬特异（类型2）的Bt菌株除了在杀虫基因上具有特异性之外,所含有的aiiA、InhA基因也有显著特点。相关研究为进一步对地下害虫蛴螬高效Bt菌株的改良提供了新思路。