亚硝基胍对苏芸金杆菌晶体和芽孢的影响

苏芸金杆菌(Bacillus thuringiensis)作为高效、无公害农药用于防治农、林、果、蔬害虫取得了良好的效果。近年来,为了提高苏芸金杆菌对昆虫的致病力,国内外不少学者研究了苏芸金杆菌的伴孢晶体与杀虫毒力的密切关系,并通过自然筛选或遗传育种方法不断获     

我国部分地区土壤中的苏芸金芽孢杆菌和球形芽孢杆菌

从云南、贵州、四川和陕西4省的土壤中分离到大量苏芸金芽孢杆菌(Bacillus thuringie—nsis)和球形芽孢杆菌(Bacillus sphaeticus)菌株。血清型分析表明,苏芸金芽孢杆菌分离株分属于23个血清型中的13个血清型,另有近20％的自凝型菌株及部分与所有标准菌抗血清无反应的菌株。对该两种昆虫病原细菌的生态分布规律进行了分析。研究了全部苏芸金芽孢杆菌分离株对鳞翅目、鞘翅目及双翅目的6种昆虫的毒力特性、伴孢晶体与芽孢的形态,以及晶体蛋白质成分。观察和测定了球形芽孢杆菌分离株的形态和毒力,并分析了部分菌株的晶体蛋白质成分。得到22株高效苏芸金芽孢杆菌和2株高效球形芽孢杆菌。证明苏芸金芽孢杆菌是典型的土壤微生物类群,我国西南地区土壤中的苏芸金芽孢杆菌资源十分丰富。     

苏芸金杆菌五个菌株的晶体蛋白研究

六十年代以来,国外在利用电泳方法研究苏芸金杆菌(Bacillus thuringiensis)不同菌株的致病机理方面,做了许多工作。Lecadet曾用含尿素的凝胶电泳和纸电泳研究了血清型Ⅰ的B.t.芽孢和晶体。Somerville等以Alesti为材料,用聚丙烯酰胺凝胶电泳进行研究。Cooksey以Galleriae、Sotto、Morris、Tolworthi为材料,用圆盘电泳法研究了提取毒蛋白(芽孢、晶体混合物)、晶体蛋白毒素和晶体蛋白溶解物。Glation以B.T.Berliner为材料,用Davis与SDS电泳法进行研究。本文主要报道了用聚丙烯酰胺凝胶圆盘电泳研究的苏芸金杆菌不同菌株晶体蛋白的图谱。     

苏芸金杆菌的毒力比较试验

苏芸金杆菌(Bacillus thuringiensis)不同品系对某一种昆虫,或苏芸金杆菌的某一品系对不同种昆虫表现出毒力的差别已有报道,例如血清型H_5a5b的蜡螟杆菌对蜡螟(Galleria mellonella)的毒力比H_3a3b的HD-1菌株高150倍,但对烟草夜蛾(Heliothis virescens)的毒     

亚硝基呱对苏芸金杆菌晶体和芽抱的影响

苏芸金杆菌(Bacillus thuringiensis）作为高 效、无公害农药用于防治农、林、果、蔬害虫取得 了良好的效果^[1]。近年来,为了提高苏芸金杆 菌对昆虫的致病力,国内外不少学者研究了 苏芸金杆菌的伴袍晶体与杀虫毒力的密切关 系^[2],并通过自然筛选或遗传育种方法不断获 得了新的变种和高毒效菌株^[3,4,10,11]。我国1975 年从美国引进B. thuringiensis var. kurstaki (HD-1)以来,已较广泛地应用于生产^[5],但对 原有菌种进行改良尚无报道。我们用化学诱变 剂亚硝基孤(NTG）处理HD-1菌种,获得了一 些诱变菌株,并研究了NTG 对苏芸金杆菌晶 体、芽抱以及诱变后对昆虫毒力     

乳化剂和保存条件对两种苏芸金杆菌杀蚊δ—内毒素的形响

我们已研究过有关化学物质及保存条件对苏芸金杆菌杀鳞翅目δ-内毒素的影响,本研究的目的在于考查乳化剂及保存温度与时间对两种杀蚊δ-内毒素毒力的影响。一、材料与方法 1、供试昆虫致倦库蚊(Culex fatigans)。 2、苏芸金杆菌菌株IPS—82(Bcillus thuringiensis var israelensis)L14(未鉴定)。 3、伴孢晶体的分离提纯以液体双相法结合蔗糖密度梯度法进行,提纯的晶体悬液于-20℃保存备用。     

用三种不同方法生产的BT(库斯塔克变种)制剂及其对家蚕的毒力

一前言在微生物杀虫剂中,苏芸金杆菌是最成功的商品制剂。活孢子和足量的σ-內毒素是该产品主要的毒素成分。众所周知,产品的毒力是根据菌株(不同菌株及同一菌株的不同株系)、培养基和生长条件的不同而异(Dulmage and Rhodes,1971)。但是,使用相同菌株在体外和体内条件下的培养物所进行的毒性比较尚未见报道。最近,我们用苏芸金杆菌(简称     

板状电泳在苏芸金芽孢杆菌亚种鉴定上的应用

作者用聚丙烯酰胺凝胶板状电泳技术,对1986年以前发表的苏芸金杆菌22个血清型29个亚种进行了酯酶分析,获得了较圆盘电泳更为准确、迅速、易区分的酮酶分析结果。其中H_1—H_(14)的20个亚种与圆盘电泳的结果相同。本文报道了1978—1986年间所发表的苏芸金杆菌9个亚种板状电泳的酮酶分析和生化测定的结果。从而确立了以酮酶型为主结合生化特性对苏芸金杆菌未知菌株进行鉴定的可行方法。     

苏芸金杆菌的一个新血清型

从土壤中分离出一株产生伴孢晶体的芽孢杆菌。该菌株具有苏芸金杆菌的典型特征,严生不规则型晶体,鞭毛抗原及生化反应与已知的苏芸金杆菌21个血清型不㈤,不产生β-外毒素。对棉铃虫、粘虫、大蜡螟及尖音库蚊幼虫均无毒性,是一个新血清型(H22),定名为苏芸金杆菌山东变种(Bacillus thuringiensis serovar shandongiensis, H22)。     

MSO7A防治松毛虫的效果及其与HD-1、7216的比较

MSO_7A是1981年从松毛虫(Dendrolimus)死蛹体内筛选的一株苏芸金杆菌(Bccillus thuringiensis)新的野生菌株,简称7A。该菌株具有var Kurstaki的典型特征。对松毛虫的毒力LC_(50)是HD-1的3倍以上,是7216的5倍以上。林间用药量比HD-1节者4/5。     

土壤来源的五个苏云金芽孢杆菌新亚种的鉴定

从中国土壤中分离的大量苏云金芽孢杆菌菌株中鉴定出H42、H43、H56、H60及H62等5种新H血清型,并进行了形态、培养特征、生化反应、晶体蛋白质成分及毒力特性等项检测鉴定,鉴定了5个苏云金芽孢杆菌新亚种： Bacillus thuringiensis subsp. jinghongiensis (H42), B.thuringiensis subsp. guiyangiensis (H43),B.thuringiensis subsp. rongseni(H56),B.thuringiensis subsp. pingluonsis(H60)及B.thuringiensis subsp. zhaodongensis(H62) 。毒力生物测定证明5个新亚种的代表菌株对棉铃虫(Heliothis armigera)\,小菜蛾(Plutella xylostella)\,柳蓝叶甲(Plagiodera versicolora)幼虫均无毒力。H42、H43、H56、H60对埃及伊蚊(Aedes aegypti)\,斑须按蚊(Anopheles stephensi)及尖音库蚊(Culex pipiens)亦均无毒;H62对埃及伊蚊无毒,但对尖音库蚊与斑须按蚊有低毒。     

我国森林土壤中苏云金芽孢杆菌生态分布的研究

从我国8个森林立地带(寒温带、中温带、暖温带、北亚热带、中亚热带、南亚热带、高原亚热带、热带)所属的13个自然保护区,采集了0—5cm土层林下土壤样品384个.测定了土壤pH、水分和养分.从中分离观察芽孢杆菌菌落1873个,分离出苏云金芽孢杆菌79株,并对其所属亚种进行了初步鉴定.其平均出土率和分离率分别为14.32%和4.21%.研究了芽孢杆菌和苏云金芽孢杆菌在森林土壤中生态分布的规律及苏云金芽孢杆菌对6种昆虫的室内毒力测定,从中筛选出不少的高效菌株.为研究苏云金芽孢杆菌在我国森林生态系中资源的保护、开发和利用,具有重要意义.     

金矿床区蜡状芽孢杆菌孢壁蛋白SDS-PAGE图谱及聚类分析

用SDS PAGE方法对 5株从我国金矿床区采集到的典型的蜡状芽孢杆菌 (Bacilluscereus)C6,C14 ,B2 ,JY I T1,JY X T9的孢壁蛋白同标准株AS1.12 6的孢壁蛋白共同进行比较分析 ,计算出了各蛋白电泳带的近似分子量 ,又以它们的迁移距离为标准同苏云金芽孢杆菌 (Bacillusthuringiensis)的孢壁蛋白相比较 ,得到聚类分析树状图谱 ,表明具有聚金作用的蜡状芽孢杆菌在分类学上具有相似性 ,而与具有杀虫作用的Bt菌株在分类学上则相差较远。     

Transformation of Bacillus thuringiensis vegetative cells by electroporation

A highly efficient procedure for the transformation of Bacillus thuringiensis and Bacillus subtilis using covalently closed circular plasmid DNA was developed by using the small Staphylococcus aureus plasmid pC194 and electroporation. We have achieved transformation efficiencies in B. thuringiensis subsp. kurstaki (HD-73) greater than 5 x 10(6) transformants/micrograms plasmid DNA. The electro-transformation (or electroporation) procedure also worked with B. subtilis 168 although at a 200-fold less level of efficiency. The results indicated that the plasmid exists in double and single-stranded forms both in B. subtilis and B. thuringiensis. A second single-stranded species was also observed in both species. This technique may prove to be applicable to other members of the genus Bacillus.     

Purification, amino acid sequence and characterization of Bacthuricin F4, a new bacteriocin produced by Bacillus thuringiensis

AIMS: Purification and characterization of a new bacteriocin, Bacthuricin F4 of Bacillus thuringiensis. METHODS AND RESULTS: A newly isolated B. thuringiensis subsp. kurstaki strain BUPM4, was shown to produce a novel bacteriocin named Bacthuricin F4. The highest bacteriocin activity was found in the growth medium and evidenced in the late exponential growth phase. Bacthuricin F4 could be purified by a two-step procedure: ammonium sulphate precipitation of protein from culture supernatant followed by a reverse phase chromatography. Upon purification, the specific activity was increased 100-fold. This bacteriocin was heat-stable up to 70 degrees C and resisted up to pH 3.0. Bacthuricin F4 was sensitive to proteases demonstrating its proteinaceous nature. Its molecular mass, determined by mass spectrometry was 3160.05 Da. Direct N-terminal sequencing of Bacthuricin F4 revealed the following sequence: DWTXWSXL. The latter was unique in the databases. Bacthuricin F4 was active against Bacillus species while it had little or no effect on Gram-negative bacteria. CONCLUSIONS: A strain BUPM4 of B. thuringiensis subsp. kurstaki, was shown to produce a new bacteriocin named Bacthuricin F4 of both new molecular mass (3160.05 Da) and new amino acid terminal sequence. This is, to our knowledge, the first bacteriocin exhibiting such characteristics reported to be produced by B. thuringiensis. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacteriocin produced by the B. thuringiensis strain BUPM4 respond to both criteria of thermostability and stability to low pHs. Thus, it could be used for the control of the related species of Bacillus harmful for agricultural products.     

Identification and characterization of strains of Bacillus thuringiensis by genomic fingerprinting using biotinylated phage M13 DNA]

Genomic DNA of the entomopathogenic bacterium Bacillus thuringiensis was analyzed by the genomic fingerprinting technique. The biotin-labeled single-stranded DNA of the phage M13 was used as a marker of hypervariable sequences. A procedure for analyzing the differentiation among various Bacillus thuringiensis strains was developed. Characteristic patterns of fingerprints were obtained for several strains, the main representatives of subspecies that are most frequently used in the manufacture of bacterial insecticides, such as subsp. thuringiensis, subsp. kurstaki, and subsp. galleriae. Because no essential differences were revealed in band patterns upon comparing fingerprints of crystal-producing bacterial strains with those of acrystallic mutants, it was assumed that the loss of crystal-producing ability in the insect pathogen Bacillus thuringiensis is not connected with significant rearrangement of its genome.     

Characterization and pathogenic evaluation of Bacillus thuringiensis and Bacillus sphaericus isolates from Argentinean soils

Eighty soil samples of different origin (from urban, agricultural, forested and horticultural areas) which had not previously been treated with bioinsecticides, were collected and examined to investigate the presence of Bacillus thuringiensis and B. sphaericus. From a total of 1473 bacterial isolates examined by differential staining techniques and growth on nutrient agar with the addition of penicillin and streptomycin, 31 (2.1%) strains of Bacillus sphaericus and 25 (1.6%) strains of Bacillus thuringiensis were isolated. These strains were tested for their pathogenicity against Diptera (Culex quinquefasciatus) and Lepidoptera (Anticarsia gemmatalis and Spodoptera frugiperda). Seven strains of Bacillus thuringiensis subspecies kurstaki were found to be pathogenic to Spodoptera frugiperda and twenty-two strains showed a pathological effect against Anticarsia gemmatalis. None of the strains of Bacillus thuringiensis nor the Bacillus sphaericus investigated, showed pathogenic activity against Culex quinquefasciatus. The strains of Bacillus thuringiensis were characterized serologically as belonging to six serotypes (darmstadiensis, entomocidus, kurstaki, muju, sotto and xianguangiensis). One strain seemed to be a new serotype. The electrophoretic profiles of the strains of Bacillus thruringiensis showed bands of 130 kDa similar to those found in strains pathogenic against Lepidoptera. Some physicochemical characteristics were also studied in the soil samples, in order to relate them to the presence or absence of these Bacillus species.     

苏云金芽胞杆菌转座因子的转座机理及其与杀虫晶体蛋白基因的关系

各种苏云金芽胞杆菌在杀虫毒力和杀虫谱上有很大差异。研究表明,这种特异性的杀虫毒力与存在于苏云金芽胞杆菌内的转座因子有密切关系,不同类型的转座因子其转座方式各异,总的来说可分为３种,即同源重组、转座重组和特异位点重组。这种转座过程的发生往往伴随着苏云金芽胞杆菌杀虫晶体蛋白的变异,这在基因工程菌的构建和杀虫多样性的研究上有着重要意义。     

Four New Bacteriophages of Bacillus thuringiensis

S ummary . The morphologies of 4 new bacteriophages active against various strains of Bacillus thuringiensis have been examined by electron microscopy. The sensitivity patterns to these bacteriophages of B. thuringiensis strains and of other Bacillus spp. are described.     

A new group of parasporal inclusions encoded by the S-layer gene of Bacillus thuringiensis

Bacillus thuringiensis produces various groups of active proteins, such as Cyt, Vip and Parasporin, in addition to the Cry protein. In this study we show S-layer proteins to be a new group of parasporal inclusions of B. thuringiensis . The S-layer consists of a two-dimensional lattice structure and is the outermost component of many archaeobacteria and eubacteria. The parasporal inclusion of B. thuringiensis strain CTC was found to be not a typical crystal protein encoded by the cry gene, but a proteinaceous inclusion encoded by the S-layer gene. Furthermore, the CTC-like strains (with their parasporal inclusions coded by the S-layer gene) are widely distributed and accounted for 25.4% of the B. thuringiensis strains tested. These strains constitue a new group of parasporal inclusions encoded by the S-layer gene of B. thuringiensis and shed new light on B. thuringiensis nontoxic strains.