苏云菌芽胞杆菌毒蛋白基因导入甘蓝获得抗虫转基因植株

苏云菌芽胞杆菌（Ｂｔ）毒蛋白对鳞翅目等昆虫有良好的特异性抗虫效果，转Ｂｔ基因为害虫防治和环境保护开辟了一条全新的途径。本试验采用根癌农杆菌介导的方法，成功地将Ｂｔ毒蛋白基因ＧＦＭＣｒｙＩＡ导入到甘蓝中，并获得了抗虫转基因植株。１材料与方法１．１植物...

苏云金芽孢杆菌工程菌BM B_(20-4)的杀虫晶体蛋白检测及毒力测定

苏云金芽孢杆菌的杀虫晶体蛋白由单基因编码,经过遗传操作,构建成BM B 20-4工程菌.本研究对工程菌BM B 20-4进行了杀虫晶体蛋白检测和发酵液毒力测定.结果表明:工程菌BM B 20-4杀虫晶体蛋白质含量比天然菌株提高44%,毒力效价显著提高,对小菜蛾提高了0.72倍,对棉铃虫提高了0.75倍.

Bt8010菌电镜观察、生化反应及对家蚕的致病力测定

本文描述了苏云金杆菌8010菌株(简称Bt8010)营养体、芽胞和伴胞晶体在扫描电子显微镜(SEM)下所看到的形态特征,并讨论了伴胞晶体的形态与其杀虫特异性的关系;再对8010菌的生化反应特性进行了鉴定与比较分析;并以4—5龄家蚕幼虫为供试虫,对来自瑞士、美国及我国湖北、河北和福建省的6种苏云菌杆菌制剂进行室内致病力的测定与比较。

Protein Characteristics and Field Efficacy of YN1-1, a Highly Virulent Strain of Bacillus thuringiensis

[Objective] This study aimed to investigate the biological characteristics of a Bacillus thuringiensis strain YNI-1, which has high virulence to Lepidoptera spp. [Method] The crystal protein of YNI-1 was analyzed by SDS-PAGE, and its indoor and field efficacy for Lepidoptera spp. was investigated. [Result] The parasporal crystal of YNI-1 has a diamond-like structure. The molecular weight of the original toxin protein is 136 kDa. After trypsin treatment, the original toxin protein was hy- drolyzed into active toxin protein with molecular weight of 63 kDa. For Plutella xy- Iostella and Pieris rapae, the indoor efficacy of B. thuringiensis was better than that of commercial B. thuringiensis （WP）. In view of field efficacy, rate of YNI-1 strain was higher than that of commercial B [Conelusion] YNI-1 strain has excellent development potential. the insects reduced thuringiensis （WP）.

Properties study and phylogenetic analysis of a bacterial strain with high de-emulsification efficiency

For further practical application, the phylogenetic analysis and de-emulsification properties study of strain XH1 with high de-emulsification efficiency isolated from crude oil contaminated soil in Daqing oil field were conducted with a surfactant-stabilized water-kerosene model emulsion. The factors influencing the de-emulsification efficiency and the generation site of de-emulsification active component of the strain were also investigated. The similarity of 16SrDNA sequences between strain XH1 and Bacillus mojavensis （DQ993678）was 99%. According to the physiological biochemical test, strain XH1 was preliminarily identified as Bacillus mojavensis. The logarithmic growth, stable phase and decline phase of strain XHI were determined as 14, 18 and 28 h, respectively. The best de-emulsification activity emerged after cultivating for 18h, and the complete de-emulsification was achieved at 24 h. The most favorable incubation conditions for de-emulsification occurred with pH of 6. 0 at 30 ℃. The de-emulsification capability of strain XH1 was mainly resulted from extracellular metabolites. The above results indicate that strain XH1 has high de-emulsification efficiency and is potential as a commercial de-emulsifier.

The Impact of Beauveria bassiana, Bacillus thuringiensis （Bt） Spray, Trichogramma pretiosum and Spinosad on the Lepidoptera： Crambidae, European Corn Borer （Ostrinia nubilalis）

The research cxamined the effects of microbial and novel insecticidal control strategies and also Irichogramma pretiosum on the I,epidoptera： Crambidae, the European corn borer （ECB）, （Ostrinia nubilalis）. The treatments included ~ino^d, from bacteria the entomopathogenic fungus Beauveria bassiana, the egg parasitoid Trichogramma pretiosum, and foliar applied Bt spray, and t^vo combinations Bearveria bassiana plus Bt spray and Trichogramma pretiosum and spinosad. The infestations by the European corn borer larvae on stalks of corn were very＂ severe in all the control treatments both 2006 and 2007. The microbial treatments caused various levels of European corn borer mortality. In all the parameters there was a significant difference between the control and other treatments with a P 〈 0.00 l. In this research, treatment with spinosad emerged as the most effective biological agent in the control of the European corn borer. The treatments had no significant effects on the relative abundance and composition of non-target arthropods （P 〉 0.05）. This research is relevant in boosting underutilized control strategies and increasing stakeholder adoption of integrated pest management practices and thereby＇ reducing the use of conventional insecticides especially for orgamc thrmers. This is very important especially in certain countries where the commumities are skeptical about consuming transgenic crops.

A New Strain of Entomopathogenic Bacillus of the Tea Pest, Eterusia magnifica （Butl.） （Zyganiidae： Lepidoptera） from Darjeeling Foothill Region of India

An entomopathogenic strain of Bacillus sp. was isolated from diseased red slug caterpillars of the leaf-feediug pest of tea, Eterusia magnifica, from the Darjeeling foothill region. Analysis of the bacterimn based on polyphasic approach such as growth phase, biochemical tests, whole body＂ protein, crystal protein profiles along with bioassay （i.e. LC50 and LT50 values） established it as a different strain but close to Bacillus thuringiensis kurstaki （Btk）, the commercial microbial pesticides of lepidopterans. Among biochemical parameters differences were noted between the new strain and Btk in ONPG, lysine decarboxylase, omithin decarboxylase, urease, nitrate reduction, V-P and glucose utilization tests. PAGE analysis of the whole body protein for the new strain recorded a 34 kDa band which was absent in Btk （used as reference）. Crystal protein profile of the newly isolated bacterial strain showed 53 and 49 kDa bands whereas in Btk only 52 kDa band was evident. Although the LC50 values of the new strain and Btk were close, their LT50 values were much different, the new strain showing a lower value than Btk. In light of the above differences and in absence of any report of entomopathogenic bacterial strain of E. magnifica, the isolated strain of Bacillus appeared to be new to science and hence was designated as RS01. The new strain opens up the possibility of its futttre use as microbial pesticide after standardizing its formulation and checking its safety aspects.

General Evaluation of Biocide （Bt-ASF-1） Produced from Iraqi Isolate of Bacillus thuringiensis

General evaluation of isolate Bacillus thuringiensis （Bt-ASF-1） used as biocide in meddle scale application was conducted. Some morphological and confirmation tests were achieved. The sensitivity tests had been accomplished by diffusion and dilution techniques to determine the response of isolate against the antibiotics. The results of diffusion tests showed to the sensitivity of bacteria to antibiotics of cefixime, erythromycin, gentamicin and tetracycline respectively. It was resistant to trimethoprim sulfonamide （TMP）, bacitracin, penicillin and all its generations, and moderate resistance to nalidixic acid. Minimum Inhibitory Concentration （MIC） for amoxicillin was ranged between 30-40 pg/mL and these results are an approximation of the universal findings. Curing experiments showed the effective role of sodium dodecyl sulfate （SDS） （1.5%） comparing with temperature. The bacterial cells became sensitive to amoxicillin and TMP. The curing by temperature did not differ significantly from control treatment in plasmid pattern or antibiotics response. Plasmid profile referring that curing by SDS has been caused disturbance in beta -lactamase genes through the sensitivity to amoxicillin and remaining resistance to ampicillin. Curing isolate by SDS also became more sensitive to nalidixic acid, erythromycin and tetracycline respectively. It was found from the curing treatments the complexity distribution of r-genes between different plasmid size and chromosome but not effect on their insecticidal ability.

Risk Assessment and Biosafety Studies of Transgenic Bt Rice （Oryza sativa L.）

Transgenic crops having alien genes from different sources are getting popularity. A Rice （Oryza sativa L.） containing gene from Bacillus thuringiensis, a soil bacterium, was assessed to study the potential risks of transgenic plants on environment. The crop was found resistant to target insect pests. Rice variety Basmati-370 transformed with two insecticidal genes, crylAc and cry2A, was grown under field conditions for several years. Data were collected at different stages of the plant growth. Fate of Cry protein in soil, effect of Bt protein on non-target insects, risks of vertical and horizontal gene flow were evaluated. No potential hazard was found at all levels. Bt protein was unstable and degraded significantly in soil within 30 days after harvesting the crop. No harmful effects were found on non-target insects （insects other than order lepidoptera）. Maximum gene flow of 0.02% was observed at close spacing and no evidence of horizontal gene transfer to Rhizobium spp. was found. In conclusion, the transgenic rice plants transformed with Bt genes have no harmful effects on the environment.

Aerobic Cr(VI) Reduction by an Indigenous Soil Isolate Bacillus thuringiensis BRC-ZYR2

Chromium(Cr) may cause losses in the yield of field plant, which is one of the favorite habitats of Bacillus thuringiensis(Bt). The purposes of our study were to assess the Cr(VI)-resistance and Cr(VI)-reducing abilities of an indigenous soil isolate of Bt and to determine the factors governing Cr(VI) reduction. Towards this end a novel dichromate-reducing Bt BRC-ZYR2, characterized with insecticidal crystal proteins(ICPs), was isolated from a uranium deposit. Minimum inhibitory concentrations(MICs) of Cr(VI) were determined by broth dilution method and the concentrations of Cr(VI) and total Cr in the supernatant were quantified colorimetrically using 1,5-diphenylcarbazide(DPC) reagent and a mixture of sulfuric-nitric acids, respectively. The isolate contained five ICP genes(cry1Ba, cry1 Bb, cry1Be/cry1 Bf, cry9 Ca and cry9Da) and exhibited a high level of Cr(VI) resistance with MICs of 150 mg L-1at pH 7.0 and 30?C, and 500 mg L-1under optimal conditions(pH 9.0 and 40?C). The total Cr concentration was similar to initial concentration of Cr(VI) under the optimal condition, suggesting that the essential removal of the Cr(VI) was dependent on Bt reduction. Under optimal conditions, the initial Cr(VI) concentrations from 25 to 75 mg L-1significantly decreased in 24 h after incubation. Addition of Mn2+, Co2+, Mo2+and Cu2+activated Bt-mediated Cr(VI) reduction, while Zn2+, Ni2+and glucose were found to inhibit the reduction. Our results indicated that this isolate could be a promising biopesticide with the potential for both insect biocontrol and Cr bioremediation in the field.