苏云金芽孢杆菌防治柑桔害虫的后效作用

1985年在广西隆安县从未施过Bt菌剂的柑桔园中喷洒Bt菌剂(Bt5a5b).1986年对该园的土壤和柑桔叶片取样分析,结果表明,施菌区土壤及叶片的Bt(H5a5b)检出率均高于不施菌剂的对照区。1987年检测果园的鳞翅目害虫,施菌区活虫的Bt检出率为51.4％,其中Bt(H5a5b) 占60.2％,死虫的Bt检出率为81.3％,其中Bt(H5a5b)占67.1％。对照区活虫Bt检出率为20.5％,其中Bt(H5a5b)占34.3％,死虫的Bt检出率为35.7％,其中Bt(H5a5b)占10％。说明施用菌剂两年后,果园中仍残留一定量的Bt,对害虫有致死的作用。     

杀线虫苏云金芽胞杆菌菌株YC-10的分离鉴定及活性测定

自烟草中分离获得对南方根结线虫Meloidogyneincognita具有较高杀线活性的拮抗细菌菌株YC-10。研究发现该菌株有伴胞晶体产生,菌体、芽胞形态及生理生化特性与苏云金芽胞杆菌库斯塔克亚种B．thuringiensissubsp．kurstaki相似,16srDNA序列分析表明其与苏云金芽胞杆菌相似性最高,初步确定该菌株为苏云金芽胞杆菌。菌株发酵液及伴胞晶体蛋白对南方根结线虫J2有很强的杀线活性,发酵液处理南方根结线虫,24h线虫死亡率为100％;伴胞晶体蛋白处理南方根结线虫,24h的LC50为5．86μg．mL^-1。研究结果显示该菌株在线虫生物防治上有较大的应用潜力。     

对甜菜夜蛾具有杀虫活性的Bt菌株cry1Ca基因研究

根据已报道的14个cry1Ca基因,设计能够扩增cry1Ca全长基因的简并引物。利用该引物对本实验室分离的472株Bt菌株进行筛选。PCR扩增发现22株菌含有cry1Ca基因。对22株菌株进行cry基因多样性分析,结果获得5种类型酶切图谱,表明这些菌株分为5种类型。SDS-PAGE结果显示22株菌均表达约130ku的蛋白。Western Blotting结果证实这些株菌中cry1Ca基因均正常表达。提取菌株的晶体蛋白,经胰蛋白酶活化,对甜菜夜蛾[Spodoptera exigua(Hübner)]进行初步生测,结果表明22株菌中活化的Cry蛋白对甜菜夜蛾均表现出很强的毒杀作用。进一步从5个类型的菌中各挑选一株毒力较高的菌株进行LC_(50)的测定,结果证实它们均具有很高毒力,其中,T1-E12(0.087μg/g)、B16-C8(0.103μg/g)、T1-B8(0.202μg/g)的活性与阳性对照菌株G03(0.090μg/g)相当,具有生产应用潜力。本研究为新型高效Bt菌株的挖掘奠定了基础。     

Laboratory and field tests of spray-dried and granular formulations of a Bacillus thuringiensis strain with insecticidal activity against the sugarcane borer

Formulations of Bacillus thuringiensis Berliner (Bt) with insecticidal activity against the sugarcane borer, Diatraea saccharalis Fabricius (Lepidoptera: Pyralidae), were developed and tested under laboratory and field conditions. The formulations were prepared using biodegradable polymers such as modified corn starch as an encapsulating agent, gelatin as an adherent, powdered sugarcane as a feeding stimulant and a Bt var. kurstaki GM-34 strain from a non-sugarcane region as the active ingredient. The spore-crystal complex of this strain was mixed at three different concentrations (30, 70 and 100 g kg(-1)) with the other ingredients. The blends were prepared as spray-dried and granular formulations, and then submitted to laboratory tests with two day old larvae of D. saccharalis and field tests in sugarcane crops with natural sugarcane borer infestation. Spray-dried formulations in laboratory bioassays caused mortality near 100% with all three concentrations, and granular formulations caused mortality around 84%. The field tests showed that spray-dried formulations at 70 and 100 g kg(-1) concentrations were as effective as a commercial bioinsectide (Lepinox), while granular formulations were ineffective.     

小菜蛾对苏云金杆菌制剂的抗药性及其对策

作者首次报道了我国部分地区小菜蛾对Ｂｔ制剂的抗药性问题，华南、广州、深圳一带小菜蛾对Ｂｔ制剂的抗性比扬州小菜蛾强１０～２５倍，平均１７倍，而泰国曼谷小菜蛾的抗性又比华南的抗性小菜蛾增强４０％左右。抗性小菜蛾对Ｈ１、Ｈ３、Ｈ５、Ｈ７４个常用血清型的参试菌株均较敏感；试验结果还表明，小菜蛾在Ｂｔ与化学农药之间无交互抗性。Ｂｔ与化学农药复方制剂对小菜蛾有较好的防效，已在深圳、广州等地示范推广死亡率达８１．５～９０．９％。对防治抗Ｂｔ害虫的对策本文作了讨论     

Bt杀虫蛋白对棉铃虫的抗虫毒力和增效作用

用 Bt杀虫蛋白 (ICP) Cry1A汰选的棉铃虫 (H elicoverpa armigera)抗性种群和同源对照种群分别测定了 4种不同类型 Bt ICP的抗虫毒力。结果表明 :4种杀虫蛋白对对照种群的毒力顺序为 :Cry1Ac>Cry1Ac+ 1C>Cry2 A>Cry1C;对抗性种群的毒力顺序为 :Cry1Ac+ 1C>Cry1Ac>Cry2 A>Cry1C,其中Cry1Ac+ 1C表现出对抗性种群有显著的协同增效作用。采用从 Bt液体发酵上清液中提取的增效粉 ,与Cry1Ac以 1∶ 1和 1∶ 2的比例混合 ,对 Cry1Ac有显著的增效作用 ,用抗性种群测定的增效比值明显大于对照种群 ,处理 5d的增效比值大于 14d的     

苏云金芽胞杆菌G033A对新发南美番茄潜叶蛾的室内毒力及田间防效

南美番茄潜叶蛾Tuta absoluta（Meyrick）是新近传入我国的一种世界毁灭性番茄害虫,对我国番茄产业的潜在威胁巨大。为有效控制南美番茄潜叶蛾,测定了新型苏云金芽胞杆菌基因工程菌G033A（Bt G033A）对该昆虫的室内毒力及田间防效。结果表明,在室内条件下,以32000 IU/mg Bt G033A WP处理过的番茄叶片饲喂南美番茄潜叶蛾幼虫,其室内毒力效果由高至底依次为4龄幼虫,3龄幼虫,1～2龄幼虫,致死中浓度LC₅₀分别为14.63、15.59和23.17 g/L;浓度在10 g/L及以上时,Bt G033A对南美番茄潜叶蛾各龄幼虫的毒力均较高,处理后96 h（第4 d）校正死亡率均超过90%。在田间条件下,以Bt G033A WP 100倍液（10 g/L）喷雾防治低龄幼虫即有较好的防效,药后第5 d对1龄和2龄幼虫的校正死亡率分别为89.1%和98.0%,药后第7 d对1龄和2龄幼虫的校正死亡率均为100%;并且,与常用的生物杀虫药剂鱼藤酮的防效相当。研究结果对有机蔬菜生产基地新发南美番茄潜叶蛾的有效控制及其综合防控方案制定,具有重要指导意义。     

Inheritance of resistance and cross-resistance to deltamethrin in Plutella xylostella (Lepidoptera: Plutellidae) from Pakistan

A field population of Plutella xylostella (L) from Pakistan was found to be highly resistant to deltamethrin (>500-fold) but had little or no resistance to spinosad, fipronil, indoxacarb, abamectin, Bacillus thuringiensis (Bt) var kurstaki, Bt var aizawai or Cry1Ac when compared with a susceptible laboratory population, Lab-UK. A sub-population was selected for six generations (laboratory G3-G8) with deltamethrin (delta-SEL), while a second sub-population was left unselected (UNSEL). Bioassays at G9 found that selection with deltamethrin gave resistance ratios of >230 compared with UNSEL (>6730 compared with Lab-UK). The delta-selected population showed no apparent cross-resistance to spinosad, fipronil or indoxacarb. Logit regression analysis of F1 reciprocal crosses between delta-SEL and Lab-UK indicated that resistance to deltamethrin in the delta-SEL population was inherited as an autosomal, incompletely dominant (D(LC) = 0.67) trait. A direct test of monogenic inheritance based on a back-cross of F1 progeny with delta-SEL suggested that resistance to deltamethrin was controlled by more than one factor.     

Can agrobiont spiders (Araneae) avoid a surface with pesticide residues?

Integrated pest management includes the use of various pesticides that can have a detrimental effect on spiders. A pesticide can express its activity directly or via residues. Most of the attention paid to the impact of pesticides on spiders has investigated the effects of direct exposure. Our aim in this study was to investigate the residual activity (repellency and toxicity) of selected pesticides on six principal species of spider occurring in an apple orchard. Four of these species (Clubiona spp., Pardosa spp., Philodromus spp., Xysticus spp.) are hunting spiders, and two species (Dictyna spp., Theridion spp.) are web-building. We were interested to see whether these spiders could avoid a surface with residues and to examine their susceptibilities to fresh and 1-day-old residues. For this purpose we conducted laboratory experiments using three types of pesticide, an organophosphate, phosalone (Zolone), a pyrethroid, permethrin (Ambush) and Bacillus thuringiensis (Bt; Novodor), which were applied to paper discs or branches. Spiders could choose to stay on any treated paper/branch within a dish. Except for Theridion spp., all species were repelled by fresh residues but not by 1-day-old residues. Clubiona spp., Dictyna spp., Pardosa spp. and Xysticus spp were repelled by phosalone and permethrin residues only, whereas Philodromus spp. specimens were repelled also by Bt residues. Theridion spp. avoided contact with residues by making a web on any available branch. Regarding toxicity, neither fresh nor 1-day-old residues of phosalone or Bt caused significant mortality to any spider species. In contrast, both fresh and 1-day-old residues of permethrin caused significant mortality to all species except for Theridion spp. In conclusion, spiders can recognise and avoid only fresh residues, although older residues might still be toxic to them.     

Abiotic factors affecting the larvicidal activity of the Bacillus thuringiensis Vip3Aa16 toxin against the lepidopteran pest Ephestia kuehniella

Journal of Plant Diseases and Protection - BUPM95 is a Bacillus thuringiensis subsp. kurstaki strain producing the Vip3Aa16 protoxin with an interesting insecticidal activity against the...     

苏云金杆菌防治棉花红铃虫的研究

室内测定苏云金杆菌对红铃虫有很高毒力,LC50为6.17—16微克/毫升。1983—1985年用工业菌剂田间试验,使铃害减少42—53％,含虫量下降24.1—95％,平均56.1％,用菌剂加少量杀灭菊酯混用有明显增效作用,防治第二,三代红铃虫效果可达70～95％,皮棉增产21—35％。防治第二代后,对第三代发生为害有较好的控制效果。     

广西苏云金杆菌资源调查研究

1978～1982年,对广两地区苏云金杆菌的资源,包括分布、种类、毒力等进行了系统调查。从不同经纬度的11个县44个点,采集土壤、虫尸样品4454份,分离出172个菌株。经过生理、生化和血清学的分析试验,作出分类鉴定;并找出广西苏云金杆菌的优势种为蜡螟变种,占94％,其毒力也是最强的。     

植物提取物对苏云金芽孢杆菌杀虫活性的影响

植物提取物对苏云金芽孢杆菌Bacillus thuringiensis(Bt)的杀虫活性有不同程度的影响。试验结果表明,处理48 h后,花椒Zanthoxylum bungeanum、假连翘Duranta repens、南洋楹Albizia falcataria、羊蹄甲Bauhinia variegata、番石榴Psidium guajava、荷花玉兰Magnolia grandiflora 的乙醇提取物对Bt的杀虫活性有显著增效作用; 小蜡Ligustrum sinense、胜红蓟Ageratum conyzoides、 一品红Euphorbia pulcherrima、烟草Nicotiana tabacum、南洋杉Araucaria cunninghamii、 繁缕Stellaria media、光叶子花Bougainvillea glabra、小飞蓬Conyza canadensis、地肤Kochia scoparia的乙醇提取物对Bt杀虫活性有显著拮抗作用;其他植物提取物则没有显著影响。     

Bt抗性和敏感棉铃虫钙黏蛋白及氨肽酶N1表达量的比较

棉铃虫体内的钙黏蛋白（CAD）和氨肽酶N1（APN1）是Bt的主要作用靶标,两种受体蛋白的变化也是棉铃虫对Bt产生抗性的主要原因。本文利用实时荧光定量PCR技术比较了Bt抗性和敏感棉铃虫CAD和APN1的表达量变化,分析受体蛋白表达量变化与抗性的关系。结果表明,与敏感品系相比,抗性品系棉铃虫CAD和APN1的表达量明显降低,1、2龄幼虫的差异达到极显著。在敏感品系中,CAD和APN1的表达量随龄期的增加呈现“V”字形变化,1、2龄幼虫的表达量最高,3、4龄时显著降低,到5龄时表达量又有所增加。抗性品系中CAD和APN1的表达量提前降低,与表达量最高的1龄幼虫相比,2、3龄时表达量显著降低,到4、5龄时又略有回升。说明棉铃虫中肠钙黏蛋白、APN1表达量的下降与抗性相关,且不同龄期棉铃虫幼虫表达量不同。     

Rapid report acetamiprid resistance and cross-resistance in the diamondback moth, Plutella xylostella

A 110-fold acetamiprid-resistant Plutella xylostella (L) strain was established after four selection experiments (in five generations) on a 9.5-fold resistant colony in the laboratory. The resistant strain did not show cross-resistance to chlorfluazuron or Bacillus thuringiensis subsp kurstaki Berliner, but displayed low resistance to cartap and phenthoate.     

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。     

苏云金杆菌(Bt)与氰戊菊酯混配防治马尾松毛虫试验

应用苏云金杆菌（Bt）对越冬代马尾松毛虫防治试验,结果表明,同一Bt浓度,室内试验幼虫死亡率较林间试验高,不同Bt浓度与1/100000的20%氰戊菊酯乳油混配林间12天防效较Bt单用有较大提高,幅度在47.96～37.35个百分点之间,林间增效作用明显,大面积防治时用500～1200倍液与1/100000的20%氰戊菊酯乳油混配为佳。     

复方苏云金杆菌商品制剂对捕食螨的影响

试验证明复方青虫菌6号商品制剂在通常稀释1000倍的剂量下(含氯氰菊酯0.5ppm),能100％杀死伪钝绥螨和智利植绥螨。不含氯氰菊酯的青虫菌制剂对两种捕食螨的死亡率分别为1.7％和7.1％。复方制剂1000倍液和500倍液在温室中施药10天后,其残效仍能使智利螨的初孵幼螨死亡率分别达到52.5％和70.7％。用含氯氰菊酯农药0.1ppm的液剂处理感染叶螨的菜豆叶,饲养智利螨雌成螨,产卵量比对照减少80％左右。为避免含有化学农药的生物制剂杀伤天敌的副作用,作者认为生物制剂中以不加化学农药为宜。必要时可根据实际情况,在使用前再加入适量的化学农药。     

苏云金素高产菌株的选育

用硫酸二乙酯(DES)处理苏云金杆菌CT-43菌株,得到了菌落形态、菌形和产生褐色素三种类型的突变株。光滑型(S-型)菌落自发突变的比例依亚种的不同而不同,CT-43菌株高达13.0％,苏云金亚种为10.6％,以色列亚种为3.9％,库斯塔克亚种不产生S-型菌落,S-型菌落依DES处理CT-43菌株芽孢时间的延长而增加,处理90分钟达49.0％。除个别S-型菌落外,均不产生苏云金素。诱变产褐色素菌落的频率为1.4％,它们都不产苏云金素。菌形变异株有仅芽孢,仅伴孢晶体,芽孢和伴孢晶体,营养体和芽孢,营养体和伴孢晶体,营养体、芽孢和伴孢晶体发生变异六种类型,它们产生苏云金素的水平各不相同。还获得了苏云金素产量比出发菌株明显提高的二个菌株。