微生物转化污泥制备苏云金杆菌生物杀虫剂

以城市污水处理厂的污泥为原料,探索了微生物转化污泥制备苏云金杆菌生物杀虫剂的可行性,并与常规培养基的发酵进程进行了对比.主要考察了苏云金杆菌的代谢特征、菌体形态与杀虫晶体蛋白产量.研究表明:无需任何预处理工序,污泥所含营养成分即可基本满足苏云金杆菌生长需求,且增殖较快,24h即可达活菌数与活芽孢数的最大值:9.48×10⁸CFU·mL^-1和8.51×10⁸CFU·mL^-1,比常规培养基提前12h,数量分别提高17%和21%;36h SEM表征显示,污泥中晶体与芽孢大部分游离,且晶体较大,呈规则的菱形,而常规培养基中苏云金杆菌的代谢进程相对滞后,且晶体较小;至发酵终点污泥中杀虫晶体蛋白含量为2.80 mg·mL^-1,亦略高于常规培养基.采用污泥发酵制备苏云金杆菌生物杀虫剂大大降低了生产成本,且发酵性能优良,为污泥处置开辟了崭新途径.     

重金属生物吸附研究中蜡状芽孢杆菌菌体微观形貌的原子力显微镜观察与表征

对蜡状芽孢杆菌在重金属生物吸附条件下细胞的界面形态及其变化情况进行了原子力显微镜成像观察与表征 .结果表明 ,在严格实验对照条件下未加入Pb(II)的空白菌体细胞呈杆状 ,单个细胞长约 3 3μm ,横截面分析结果为宽 1 9μm左右 ,高为 0 3～ 0 4 μm ,细胞壁表面比较光滑 ,细胞之间以竹节结构首尾相连排列成长杆状 ;吸附Pb(II)后细胞的体积则发生膨胀 ,单个细胞长约 3 9μm ,横截面分析结果为宽 3 1 μm左右 ,高约为 1 1 5 μm ,细胞壁表面变得比较粗糙 ,细胞之间较容易发生粘结 ,与对照空白菌体相比 ,细胞之间呈长杆状的竹节结构排列则变得很少 .采用截面分析曲线中形貌特征的高宽度比R对细菌细胞在云母基底表面的横向铺展变形作用进行了定量表征 (吸附铅离子后R值由 0 1 6 5变为 0 4 1 6 ) ,同时探讨了细胞在基底表面的横向铺展变形作用可能与细胞表面电荷影响细胞在基底表面的粘附性能密切相关 ,并受溶液pH、离子强度以及金属离子的暴露与吸附等外界因素影响而发生改变 .有关其详细机理 ,还有待在以后的研究工作中作进一步探讨     

苏云金芽孢杆菌无质粒突变株BMB171的转化和表达性能

研究了苏云金芽孢杆菌无质粒突变株ＢＭＢ１７１的转化性能和表达ｃｒｙ１Ａｃ和ｃｒｙ１Ｃａ基因的性能．用ｐＨＴ３１０１、ｐＢＭＢ３０５、ｐＢＭＢ１７３６、ｐＢＭＢ６７１、ｐＢＴＬ１和ｐＨＶ１２４９等６种外源质粒电转化无质粒突变株ＢＭＢ１７１,其转化频率分别是Ｂｔ４Ｑ７、Ｂｔ４Ｄ１０和Ｂｔｉ．ＩＰＳ．７８／１１等３种常用受体菌相应最高转化频率的１０００、６．７、１２．５、６６．７、３５００和２倍,其每μｇＤＮＡ的最高转化子数达１０７．导入ＢＭＢ１７１的外源质粒的稳定性与其复制子类型和质粒大小有关．无质粒突变株ＢＭＢ１７１表达ｃｒｙ１Ａｃ基因的表达量高于对照受体菌Ｂｔ４Ｑ７,略低于Ｂｔ４Ｄ１０和Ｂｔｉ．ＩＰＳ．７８／１１,而ＢＭＢ１７１表达ｃｒｙ１Ｃａ基因的表达量高于这３种受体菌;对小菜蛾３龄幼虫和甜菜夜蛾初孵幼虫的毒力测定结果表明,ＢＭＢ１７１表达ｃｒｙ１Ａｃ基因的杀虫毒力高于Ｂｔ４Ｑ７和Ｂｔ４Ｄ１０,略低于Ｂｔｉ．ＩＰＳ．７８／１;而表达ｃｒｙ１Ｃａ基因的毒力高于这３种受体菌     

BACILLUS SP. BT-7木聚糖酶基因克隆与鉴定

报道了从一株有较高半纤维素酶活性的Ｂａｃｉｌｕｓｓｐ．ＢＴ７克隆内切木聚糖酶基因的结果．以大肠杆菌ＸＬ１ｂｌｕｅ为宿主菌,采用水解圈检测法,在含有ＲＢＢ木聚糖（４氧甲基Ｄ葡萄糖苷Ｄ木聚糖Ｒｅｍａｚｏｌ亮蓝Ｒ）的平板上分离到能水解ＲＢＢ木聚糖的阳性克隆３个,对它们进行的限制酶作图和Ｓｏｕｔｈｅｒｎ杂交分析表明,它们属于３种不同的内切木聚糖酶基因     

Chromium phytoextraction and physiological responses of the hyperaccumulator Leersia hexandra Swartz to plant growth-promoting rhizobacterium inoculation

● Improved Cr phytoextration efficiency was achieved by B. cereus inoculation. ● B. cereus could produce plant-beneficial PGPR factors at diverse Cr stresses. ● Enhanced resistance of inoculated L. hexandra towards elevated Cr stress. ● The majority of Cr existed in the stable forms in the tissues of L. hexandra. Phytoextraction is a promising option for purifying hexavalent chromium (Cr(VI))-laden wastewater, but the long remediation period incurred by poor growth rate of Cr hyperaccumulators remains a primary hindrance to its large-scale application. In this study, we performed a hydroponic experiment to evaluate the feasibility of promoting the growth and phytoextraction efficiency of Cr hyperaccumulator Leersia hexandra Swartz (L. hexandra) by inoculating plant growth-promoting rhizobacteria (PGPR) Bacillus cereus (B. cereus). In batch tests, the Cr(VI) removal rates of L. hexandra and B. cereus co-culture were greater than the sum of their respective monocultures. This was likely due to the microbial reduction of Cr(VI) to Cr(III), which is amiable to plant uptake. Besides, the PGPR factors of B. cereus, including indoleacetic acid (IAA) production, 1-aminocyclopropane-1-carboxylic acid deamination (ACCd) activity, phosphate solubilization capacity, and siderophore production, were quantified. These PGPR factors helped explain the biomass augmentation, root elongation and enhanced Cr enrichment of the inoculated L. hexandra in pot experiments. Despite the increased Cr uptake, no aggravated oxidative damage to the cell membrane was observed in the inoculated L. hexandra. This was attributed to its capacity to confront the increased intracellular Cr stress by upregulating both the activities of antioxidative enzymes and expression of metal-binding proteins/peptides. Moreover, L. hexandra could always conserve the majority of Cr in the residual and oxalic integrated forms with low mobility and phytotoxicity, irrespective of the B. cereus inoculation. These results highlight the constructed Cr hyperaccumulator-rhizobacteria consortia as an effective candidate for decontaminating Cr(VI)-laden wastewater.     

不同陈化烟叶叶面高温微生物多样性

以国内外不同品种、不同陈化时间的10个烟叶样品为材料,对其叶面高温微生物进行了分离纯化、鉴定和多样性研究,以了解表面高温微生物在烟叶陈化发酵中的作用.结果表明:陈化烟叶表面高温菌总量为103~105个/g,高温细菌占绝对优势,真菌很少,未能分离到放线菌和酵母菌;高温细菌主要包括芽孢杆菌属(Bacillus)和地芽孢杆菌属(Geobacillus)两个属,芽孢杆菌属为优势菌属;高温霉菌主要包括青霉属(Penicillium)和曲霉属(Aspergillus),曲霉属为优势菌属.国内外烟叶表面微生物种类和数量有所差异,与陈化时间和烟叶品种等因素有着密切关系.