Expression of the Truncated vip3A Gene at the 3'-terminus from Bacillus thuringiensis in Escherichia coli

将苏云金芽孢杆菌(Bacillus thuringiensis)缺失C端154个氨基酸编码区的vip3A基因(vip3T)插入原核表达载体pQE30，构建了重组表达载体pQEvip3T，并转化大肠杆菌(Escherichia coli ) M15进行IPTG诱导表达，比较了完整的Vip3A蛋白和C端缺失的蛋白Vip3T的可溶性和杀虫活性。与Vip3A不同，融合蛋白Vip3T以不可溶的包含体形式存在，诱导表达的菌液中没有检测到可溶性Vip3T蛋白。生物测定结果表明，M15(pOTP)诱导表达的Vip3A蛋白对初孵斜纹夜蛾(Spodoptera litua)和甜菜夜蛾(S. exigua)幼虫具有较高的杀虫活性，其提纯的包含体无毒，但包含体的碱性裂解液却又恢复了对夜蛾科害虫的活性；M15(pQEvip3T)菌液、包含体及其碱性裂解液对这两种昆虫幼虫则完全无毒，说明在大肠杆菌中，Vip3A蛋白C端氨基酸可能对Vip3A蛋白的可溶性和杀虫活性具有重要的影响。     

Colonization of wheat seedling (Triticum aestivum) roots by Pseudomonas fluorescens and Bacillus subtilis

Summary Colonization patterns of Pseudomonas fluorescens and Bacillus subtilis on roots of wheat seedlings growing on water agar were studied qualitatively by replica printing and quantitatively by the plate count method. The results indicated a stronger colonization potential for P. fluorescens (up to 10⁷ cfu/cm root) than for B. subtilis (up to 10⁵ cfu/cm root). Although the numbers of both species were lower when inoculated together, the observed colonization patterns on the roots were comparable to those found with single inoculations. For none of these bacteria was active migration along the root surface in any direction observed, indicating that distal positions are reached mainly by a passive displacement on the root tip and elongating cells. Ecological implications of the observed phenomena are discussed.     

钾细菌制剂在花生上的应用研究

在种植花生时采用7．5kg／hm2作拌种肥处理，与增施KCl效果相似，可促进花生生长，增加花生有效根瘤、单株结荚数、百果重和出仁率，最终比对照增产l7．93％，每公顷投入产出比达1：40．8，具有明显的经济和生态效益。     

胨冻样芽孢杆菌(Bacillus mucilaginosus YNUCC0001)絮凝剂生产、絮凝特性及其系统发育学分析

对BacillusmucilaginosusYNUCC000116SrDNA的1481bp片段与GenBank中最相似的16个分类单位进行了比较。UPGMA,NJ,ME和MP方法构建的系统发育树显示B.mucilaginosusYNUCC0001与B.mucilaginosusHSCC1605T、B.mucilaginosus1480D及Paenibacillussp.NBT形成一个单系群分支。在50L全自动发酵罐中30℃发酵52h后,菌株YNUCC0001产生的胞外生物多聚絮凝剂(EBF)达到最大产率(粘度:3420C.P.)。在pH4.0、用量为0.25mlL-1的条件下,这种EBF对高岭土悬浊液的絮凝活性最大(99.8%);121℃高压灭菌60min后絮凝活性维持在98.6%。Hg2+,Ca2+,Mg2+,K+,Zn2+对其絮凝活性有促进作用,而Fe3+、Al3+、EDTA和Cu2+则有强烈抑制。     

Toxicity of Bacillus thuringiensis Crystal Toxins to Field Populations of Rice Leaf Folder, Cnaphalocrocis medinalis （Guenee） and Establishment of Baseline Susceptibility to Cry1Ab

Eight insecticidal crystal proteins of Bacillus thuringiensis, CrylAa, CrylAb, CrylAc, CrylB, Cry2Aa, CrylC, CrylDa and Cry 1Ea were assessed for toxicity against 1 st instar larvae of rice leaf folder, Cnaphalocrocis medinalis （Guenee） at 48 HAT and 72 HAT. Bioassay results depicted CrylAa was the most toxic （LCso 2.35 ppm） followed by CrylBa （LCso 8,50 ppm） and CrylAb （LCso 8.73 ppm） at 48 HAT, whereas, at 72 HAT CrylAb proved to be highly toxic （LC50 0.50 ppm） followed by CrylAa （LCso 4.07 ppm）, CrylAc （LCso 4,84 ppm） and CrylBa （LCso 6.42 ppm）. Toxins Cry2Aa, CrylCa, CrylDa and CrylEa did not resulted in any mortality at 48 HAT and 72 HAT, respectively. Baseline estimates for CrylAb against 1st instar larvae of C. medinalis sampled from seven geographical locations revealed variation in LC50＇s from 0.37 ppm to LC50 16.25 ppm at 48 HAT and LC50 0.50 ppm to LC50 6.49 ppm 72 HAT, respectively with relative resistance ratios of 44-fold and 13-fold at 48 HAT and 72 HAT over the susceptible population.     

枯草芽孢杆菌替代抗生素对平养和笼养肉鸡的影响研究

 将240只1日龄罗斯308（ROSS 308）肉仔鸡，按照饲养方式（地面平养和笼养）以及添加枯草芽孢杆菌替代抗生素与否（替代和不替代）分为4个处理组（1组：抗生素地面平养组；2组：抗生素笼养组；3组：枯草芽孢杆菌地面平养组；4组：枯草芽孢杆菌笼养组），进行42 d的饲养试验。结果显示：试验全期，肉鸡末体重和饲料/增重比指标，4组显著优于（P<0.05）1组，2组和3组之间无差异（P>0.05）。21日龄和42日龄肉鸡血清生化指标分析显示：4组肉鸡血清白蛋白浓度显著大于1组；且血清尿酸浓度4组小于1组；而2组和3组之间所有血清测定指标均无差异。以上结果证明，无论是地面平养还是笼养方式，枯草芽孢杆菌替代抗生素对改善肉鸡生长性能均有良好效果，其作用机理主要是提高饲料蛋白的营养利用率.     

甘蔗黑穗病菌拮抗菌HAS的筛选和鉴定

采用传统形态学鉴定及16S rRNA基因序列分析,筛选鉴定对甘蔗黑穗病菌具有拮抗作用的细菌的归属,并采用细菌的通用引物P0、P6扩增16S rRNA基因片段.结果表明:得到1 533 bp的DNA片段,其序列与枯草芽孢杆菌序列的同源性高达99％.其形态学特征:菌体为短杆状,能运动；革兰氏阳性反应；3％KOH溶解性实验呈阴性反应；有鞭毛,鞭毛周生；芽孢中生,椭圆形,孢囊稍膨大,初步将该菌株归属为枯草芽孢杆菌.与病原真菌对峙培养的结果表明,该菌对引起甘蔗和其它作物病害的多个植物病原真菌均具有很好的抑制作用.据此,可将分离的菌株HAS确定为对甘蔗黑穗病菌有较好防治效果的枯草芽孢杆菌.     

BP神经网络结合正交试验法优化Bacillus velezensis Z-27菌株培养基组分

将BP人工神经网络(Artificiai neural network)技术与传统的正交试验方法相结合,提出一种新的试验分析和处理方法,利用神经网络特有的自学能力,对主要影响因素进行仿真优化,获得Bacillus velezensis Z-27菌株培养基组分,即玉米粉2.0％、豆饼粉1.5％、MnSO4· H2O 0.07％,起始pH为7.0、接种量为2.0％,应用优化得到的培养基组分进行验证试验,取得了较好的效果.     

Changes of Morphological Parameter of Wheat Seed Pretreatment by the Biocontrol Agents Bacillus subtilis QM3 during Germination

Bacillus subtilis QM3, a potential biocontrol agent and useful plant-growth-promoting bacteria (PGRB) from yak dung in China with broad-spectrum and strong antifungal bioactivity, was studied to determine if it either stimulated or inhibited the germination of wheat seeds depending on the amount of bacteria. Germination rate (GR) and length of shoot (LS) increased at 10-fold dilution of QM3 fermentation liquid for 12 h (P < 0.05). Germination potentiality (GP) and root cap ration (RC) decreased at 100-fold dilution of QM3 for 0 h. Length of root (LR) and tips of root (TR) at day 7 remained at control levels at 0-fold dilution of fermentation liquid of QM3 for 24 h. The results obtained suggest that some plant diseases, environmental stresses (such as high salt and heavy metal contents), and difficulties in germination can be controlled or relieved by the use of QM3.     

柑橘溃疡病拮抗芽孢菌株的筛选及其控病作用

从柑橘根际土壤中获得192个细菌分离物，通过平板拮抗测试，筛选出对柑橘溃疡病菌（Xanthomonas axonopdis pv. citri）有较强拮抗作用的6个芽孢菌株，抑菌圈直径达12.0～25.0mm，其中菌株G32对柑橘溃疡病菌抑制效果最好，稳定性高；温室控病试验表明，菌株G32的控病效果最佳，提前喷施拮抗菌其预防效果达60.7%，接种溃疡病菌后喷施的治疗效果为53.2%；根据其形态特征、培养性状、生理生化特性及16S rDNA序列比对分析，将G32鉴定为枯草芽孢杆菌Bacillus subtilis。