水稻细菌性叶鞘褐腐病研究

从浙江嘉兴市和余杭市的晚稻褐腐叶鞘及谷粒的156份样本中分离出363个荧光细菌，经菌落形态和致病性测定后获得19个致病菌株，从中选出10个进行细菌学和Biolog鉴定，并与6个标准菌株比较，结果证实，这种水稻叶鞘褐腐病是由Pseudomonas fusouaginae引起，种子能带菌传病。     

Detection of Xanthomonas sp., the Causal Agent of Onion Bacterial Blight, in Onion Seeds Using a Newly Developed Semi-selective Isolation Medium

Onion bacterial blight, caused by Xanthomonas sp., is a potentially severe disease in several tropical and subtropical areas. Although little research has been undertaken on this pathosystem, seed transmission of the pathogen has been hypothesized. Because of an important bacterial microflora naturally associated with onion seeds, detection of the pathogen is difficult using non-selective agar media. A new semi-selective medium, whose selectivity was obtained by a combination of four antibiotics, was developed. The new NCTM1 medium contained (per liter) yeast extract 7g, peptone 7g, glucose 7g, agar 15g, neomycin 10mg, cephalexin 30mg, trimethoprime 3mg, pivmecillinam 100mg and propiconazole 20mg. Plating efficiencies, using 16 pure cultures of the pathogen, ranged from 79% to 142%, with an average of 110% compared to the basal medium. All onion Xanthomonas sp. strains from several countries grew on NCTM1 medium. The pathogen was repeatedly isolated using this medium from seed samples containing approximately 10⁶ saprophytic bacteria per gram, as well as from symptomless plant material. Xanthomonas sp. was detected only in seeds originating from one infected seed production site. This is the first report of selective isolation of Xanthomonas sp. from onion seeds. NCTM1 medium should be a valuable tool to study the ecology and epidemiology of Xanthomonas sp. causing onion bacterial blight.     

Pollution-induced tolerance of soil bacterial communities in meadow and forest ecosystems polluted with heavy metals

Pollution-induced community tolerance (PICT) allows finding a cause–effect relationship between pollution and adverse changes in a community. In our previous study we found that functional diversity of bacterial communities decreased significantly with increasing metal concentration, in both forest humus and meadow topsoil. Thus, the aim of the present study was to test whether tolerance of soil bacterial communities had increased as an effect of long-term metal pollution. Bacterial tolerance was tested with the use of the Biolog^® ECO plates in soils originating from the most polluted and the least polluted sites from three forest and five meadow transects located near smelters in Avonmouth (England), Clydach (Wales), and Głogów and Olkusz (Poland). We found that tolerance of bacterial communities was significantly increased in polluted meadow soils when compared to control meadow soils. On the contrary, no increase in tolerance was detected in polluted forest humus.     

秦岭山区硅酸盐细菌的分离、筛选以及初步鉴定

针对秦岭周至县不同生境的土壤进行硅酸盐细菌的分离与筛选,对20株生长较好的硅酸盐细菌进行摇瓶培养,测定其发酵液的水溶性钾含量,挑选出解钾能力较强的菌株,并使用传统生理生化特征试验和Biolog微生物自动鉴定仪两种方法对其进行鉴定.结果表明,QL21为1株解钾能力较强的菌株,解钾率为25.1%,该菌为G+革兰氏阳性,菌体为长杆状,产生椭圆芽孢以及椭圆形肥大荚膜,其生理生化特征与芽孢杆菌很接近,经Biolog微生物自动鉴定仪鉴定为芽孢杆菌.     

循环水养鳗系统生物过滤器中微生物群落的代谢特性

为了研究鳗鱼循环水养殖系统不同水处理单元的微生物群落碳代谢特征,实验采用Biolog Eco技术,分析了流化床两个槽和滴流式生物过滤器上、中、下三层的生物膜微生物群落功能多样性。结果显示,流化床两个槽和滴流式生物过滤器中、下层微生物多样性指数（Shannon-Wiener指数、Simpson指数和Pielou指数）无显著差异（P>0.05）,但均显著高于滴流式生物过滤器上层（P<0.05）。平均色度变化（AWCD）与主成分分析（PCA）均证明滴流式生物过滤器上层与其它采样点微生物代谢差异较大。各采样点微生物未对ECO板某一大类碳源表现出偏好,但对衣康酸、D-半乳糖醛酸、L-精氨酸、L-天门冬酰胺、L-丝氨酸、D-甘露醇、D-木糖、N-乙酰-D-葡萄糖氨、吐温40、吐温80、苯乙胺等单一碳源利用较好;而对γ-羟丁酸和α-丁酮酸以及D,L-α-磷酸甘油和1-磷酸葡萄糖利用较差。某些碳源种类如D-葡糖胺酸、α-D-乳糖、2-羟基苯甲酸仅能被部分采样点的微生物利用。本实验利用Biolog EcoPlateTM技术研究中试规模循环水处理单元微生物群落代谢特征,研究结果为生物过滤器的调控提供了一种新的思路即可以通过碳源调节,来促进生物膜微生物群落结构的改变以此提高水处理效率。     

奶牛源化脓隐秘杆菌的分离鉴定及其毒力基因的检测

本研究旨在分离鉴定来自北京某奶牛场死亡奶牛肺脏的1株疑似病原菌CVCC 3982,并测定其致病性。通过分离培养获得疑似病原菌,采用Biolog鉴定系统和16S rDNA序列分析对其进行了鉴定,人工接种CD-1小鼠测定了其致病性,合成引物对其主要毒力基因进行了检测。结果显示,该疑似病原菌为革兰氏阳性杆菌,β溶血,Biolog鉴定结果显示其为化脓隐秘杆菌,其16S rDNA序列与化脓隐秘杆菌模式菌株NCTC 5224的同源性达100%,系统发育分析显示其与化脓隐秘杆菌处于同一分支。腹腔注射该菌可致小鼠死亡。分离菌株基因组中含有溶血素(PLO)基因,神经氨酸酶H(NanH)基因,神经氨酸酶P(NanP)基因,菌毛基因(fimA、fimC和fimE),但缺失胶原结合蛋白(CbpA)基因和菌毛fimG基因。结果表明该分离菌株为化脓隐秘杆菌且具有致病性。     

典型血吸虫病疫区河滩地五氯酚污染土壤微生物特征——以松滋老陈镇庙河为例

采用现场采样及室内分析方法,对长期施用五氯酚(PCP)作为钉螺杀灭剂的典型血吸虫病流行疫区河滩地的土壤微生物特征进行了初步调查研究.结果表明,长期大量施用PCP对河滩裸地土壤微生物种群造成了一定程度的伤害,降低了微生物的总体活性;而种植杨树有利于恢复土壤微生物生物量碳,对修复PCP污染土壤有一定的促进作用.Biolog结果显示土壤微生物利用单一碳源能力的大小顺序为杨树林地>对照>河滩裸地,表明有机污染在一定程度上抑制了土壤微生物的生长.主成分分析显示对照、杨树林与河滩裸地的碳源利用能力差异显著,表明PCP污染对微生物的碳源利用造成了显著影响.土壤微生物对不同种类碳源利用计算结果表明,种植杨树导致林地土壤环境条件发生变化,杨树林地土壤微生物对碳源的利用也发生了明显变化.     

土壤微生物多样性的研究方法

为了更加全面有效地研究土壤微生物多样性,笔者通过对于当前国内外研究土壤微生物多样性的研究方法,包括微生物稀释平板记数法、Biolog微平板法、PLFA分析法、分子生物学方法进行了阐述,说明了几种方法操作性的优缺点,重点说明了在实验过程中应该选取的方法,为研究者选取研究方法时提供基本信息及建议。     

3株斑羚、岩羊化脓隐秘杆菌的鉴定及系统发育分析

本研究利用Biolog快速鉴定系统、16S rRNA序列分析及传统细菌鉴定方法对3株分离自北京动物园斑羚、岩羊化脓隐秘杆菌进行了鉴定和系统发育分析。结果表明,3株菌株均为化脓隐秘杆菌;系统发育分析结果表明,3株化脓隐秘杆菌16S rRNA序列与NCTC5224模式株同源性均为100%,且位于系统发育树的同一分支。     

荒漠草原不同植被类型土壤微生物群落功能多样性

利用Biolog EcoplateTM技术通过对荒漠草原生态系统中沙米、白沙蒿、柠条、沙冬青和人工乔木林5种植被类型土壤微生物群落功能多样性分析,以探讨不同植被类型对土壤微生物群落代谢功能多样性的影响.结果表明,5种植被类型土壤微生物群落代谢活性有显著的差异(P＜0.01),5种植被类型土壤微生物群落利用碳源的种类也存在差异,氨基酸类、羧酸类和糖类是其主要的碳源利用类型.在Biolog ECO板培养基接种培养96 h后,5种植被类型土壤微生物群落多样性显示,物种丰富度指数(R)和均匀度指数(EH)差异显著,Shannon 指数(H′)差异不显著.