纳米Mg(OH)_2对苏云金芽胞杆菌蛋白杀虫活性及抗紫外能力影响的研究  

The Effect of Nano-Mg(OH)_2 on Insecticidal Activity and UV Resistance of Bacillus thuringiensis Protein

作　　者：李澜[1] 陈锦灿[1] 杨兆元[1] 李东哲[1] 吴薇[1] 饶文华 潘晓鸿[1,2] 关雄[1,2] 

机构地区：[1]福建农林大学生物农药与化学生物学教育部重点实验室,福州350002 [2]闽台特色作物病虫生态防控协同创新中心,福州350002

出　　处：《农业生物技术学报》2015年第11期1452-1457,共6页Journal of Agricultural Biotechnology

基　　金：福建省教育厅高校领军人才项目(No.k8012012a);闽台特色作物病虫生态防控协同创新中心课题(闽教科[2013]51号);福建省科技计划重点项目(No.2013H0058);国家粮食局项目(No.201313002-3);福建农林大学优秀博士学位论文资助基金(No.324-1122yb003)

摘　　要：苏云金芽胞杆菌(Bacillus thuringiensis,Bt)制剂是目前产量最大、应用最广泛的微生物杀虫剂,但在施用过程中易受紫外线的影响而缩短持效期,成为Bt制剂应用受限的主要原因之一。本研究以纳米Mg(OH)2为载体,将其与Bt杀虫晶体蛋白进行负载,研究纳米Mg(OH)2负载Bt杀虫晶体蛋白后的生物杀虫活性及抗紫外能力。以共沉淀法制备纳米Mg(OH)2,利用X-射线粉末衍射(X-ray powder diffraction,XRD)对其尺寸进行表征,通过谢乐公式计算得出合成的纳米颗粒在101面的尺寸为12 nm。扫描电镜(scanning electron microscope,SEM)的结果表明,纳米颗粒为较规则的片状结构且分散均匀,与Cry蛋白负载后,其形貌没有发生明显的变化。以高效杀蚊Bt菌株LLP29为供试菌株,利用碱溶法获得Bt杀虫晶体蛋白。通过十二烷基硫酸钠聚丙烯酰胺(sodium dodecyl sulfonate-polyacrylamide gelelectrophoresis,SDS-PAGE)和生物活性测试结果表明,Bt杀虫晶体蛋白与纳米Mg(OH)2进行一定负载后,能有效地提高其Cry蛋白的杀虫活性,同时增强抗紫外辐射能力,有效地保护蛋白结构免受破坏。本研究为获得生物相容性好、环境友好的纳米Bt高效制剂提供了理论基础。Bacillusthuringiensis（Bt） is a gram-positive and spore-forming bacteria. It can form parasporal crystal during spore- forming, which is specific toxic to many insects. Bt formulation is a microbial bioinsecticide, which is the largest produced and the most widely used. But the Bt formulation is sensitive to UV in the application process, thus shortens the duration of active ingredient, which the disadvantage would limit the application of Bt formulation. Improving the biological insecticidal activity and UV-resistant ability of Bt insecticidal crystal protein by loading with nanomaterialsis is regarded as a promising technology, due to its advantages of small particle size, large specific surface area, high efficiency and low toxicity,environmental- friendly properties and improving the stability of Bt preparations. In the present study, Btinsecticidal crystal protein was loaded with nano-Mg（OH）2to understand the compound biological insecticidal activity and resistance to UV. The nano- Mg（OH）2was prepared by co- precipitation of magnesium chloride hexahydrate and sodium hydroxide, and the X-ray powder diffraction（XRD） was applied to analyze the size of synthesized nano-Mg（OH）2, and the size of nano-Mg（OH）2at（101） direction was 12 nm according to the scherrer equation. Scanning electron microscope（SEM） analysis showed that the sample contained a lot of small nano- flakes structure with uniform dispersion, and the morphology of Mg（OH）2particles loading with Cry toxin had no obvious changes. Bt LLP29 was selected as a target strain because it had high insecticidal activity to mosquitoes（Culicidae）, and the Bt insecticidal crystal protein was obtained by alkali soluble method. Results of sodium dodecyl sulfonate- polyacrylamide gelelectrophoresis（SDS- PAGE） and bioassay indicated that the loaded nano-Mg（OH）2could effectively improve the insecticidal activity of protein, protect the protein structure from destructing, and enhance the ability of resistance to ultraviolet

关 键 词：纳米MG(OH)2 Bt杀虫晶体蛋白 抗紫外能力 杀虫活性 

分 类 号：S332.3[农业科学—作物遗传育种]