连栽木麻黄根际微生物群落结构和功能特征

以不同栽培代数的木麻黄(第1代FCP、第2代SCP、第3代TCP)根际土壤为试验材料,运用BIOLOG微平板和磷脂脂肪酸(PLFA)技术分析根际土壤微生物群落结构和功能多样性对多代连栽响应。结果表明,不同代数的土壤微生物对各类碳源的利用程度存在显著差异,连栽后木麻黄根际土壤微生物对碳源利用率显著下降。在6类碳源中,除胺类外,其他5种碳源均呈现FCP>SCP>TCP。PLFA分析共检测到11种PLFA生物标记,FCP土壤微生物PLFA生物标记总量明显高于SCP和TCP,3个代数土壤中含量最高的PLFA生物标记是i16:0、a15:0和18:1ω9c。土壤中特征微生物含量差异明显,细菌分布量最大,其次是真菌和放线菌。随栽植代数增加,细菌含量减少,真菌含量增加。土壤微生物群落多样性指数均呈现FCP>TCP>SCP,与土壤理化性质变化密切相关。可见木麻黄连栽显著影响其根际土壤微生物群落结构与功能,因此根际土壤微生态失衡可能是导致木麻黄连栽障碍的重要因素。     

3种有机溶剂及其混合物对蛋白核小球藻时间毒性的探究

已有研究表明部分污染物的毒性具有时间依赖性。以蛋白核小球藻(Chlorella pyrenoidosa)为指示生物,透明96孔微板为实验载体,以3种有机溶剂甲醇(Meth)、乙醇(Eth)、二甲基亚砜(DMSO)及其不同体积比的混合物为研究对象,应用时间毒性微板分析法(简称t-MTA)系统测定有机溶剂及其混合物在0、12、24、48、72和96 h对C.pyrenoidosa的毒性效应。结果表明,有机溶剂对C.pyrenoidosa具有明显的时间依赖毒性特征,但不同有机溶剂毒性的大小不同,随时间增长速率不同,毒性大小顺序亦随时间发生变化;有机溶剂混合物也具有明显的时间依赖毒性特征,不同比例有机溶剂混合物在不同暴露时间的毒性差异不明显,但不同比例有机溶剂混合物毒性随时间的变化规律不同,在0~24 h,体积比为1:1、1:2和1:3混合物的毒性缓慢增加,2:1、3:1混合物的毒性迅速增加;在24~48 h,1:1、1:2和1:3混合物的毒性迅速增加,2:1和3:1混合物的毒性增加速率减缓;48~96h内5种比例混合物的毒性增加速率均减缓;有机溶剂混合物对蛋白核小球藻的毒性变化规律受体积比较大的组分影响。     

Genetic, phenotypic and pathogenic diversity among xanthomonads isolated from pistachio (Pistacia vera) in Australia

Repetitive extragenic palindromic polymerase chain reaction (rep-PCR), sequencing of the 16S−23S rDNA internal transcribed spacer (ITS), biochemical and physiological tests, the Biolog microplate system, polyacrylamide gel electrophoresis (PAGE) of whole-cell proteins, and pathogenicity tests were used to characterize variability among xanthomonads isolated from pistachio trees suffering from bacterial dieback in four regions of Australia. ITS sequencing and rep-PCR revealed two distinct genotypes among the strains. The ITS sequencing suggested that the pistachio strains were closely related to Xanthomonas translucens pathovars, in particular X. translucens pv . poae . Results of physiological and biochemical tests, as well as Biolog microplate analysis and protein profiling, confirmed the existence of two groups. Furthermore, pathogenicity and host-range studies indicated that the two groups were biologically different. There was an association between the two groups and the geographical origin of the strains.     

转cry1Ie基因抗虫玉米对根际微生物群落碳代谢的影响

转基因作物对土壤生态系统的影响日益引起人们的广泛关注。采用Biolog ECO微平板对转基因抗虫玉米IE09S034及其对应的受体玉米根际微生物群落碳代谢功能进行了比较研究。结果表明,转基因抗虫玉米IE09S034对根际微生物碳代谢功能无显著性影响,且发现年份和生育期是引起根际微生物碳代谢情况变化的主要因素。本文为转基因抗虫玉米IE09S034的环境安全性评估提供了新的依据。     

荧光假单胞菌与红绒盖牛肝菌共接种对杨树根际土壤酶活性及微生物多样性的影响

【目的】探究荧光假单胞菌与红绒盖牛肝菌共接种条件下NL-895杨根际土壤酶活性和土壤微生物功能多样性的变化,从微生态水平揭示溶磷细菌与外生菌根真菌互作对NL-895杨的促生机制。【方法】采用盆栽试验考察荧光假单胞菌JW-JS1与红绒盖牛肝菌(Xc)共接种后NL-895杨根际土壤酶(酸性磷酸酶、脱氢酶和转化酶)的活性变化,并通过BIOLOG微孔板法分析共接种对NL-895杨根际土壤微生物群落和功能多样性的影响。【结果】从整体趋势来看,不同接种处理后NL-895杨根际土壤酶活性均有所增强,土壤酸性磷酸酶、脱氢酶和转化酶活性均大于对照,其中,共接种JW-JS1+Xc处理对土壤酸性磷酸酶和转化酶活性的促进最为明显;接种150天后,不同接种处理土壤微生物的AWCD值均高于CK(培养24～48 h除外),且随着培养时间的延长呈先上升后稳定的趋势;培养96 h后,共接种JW-JS1+Xc处理土壤微生物McIntosh优势度指数和碳源利用丰富度指数显著高于单接种Xc或JW-JS1和CK(P<0.05),而Shannon丰富度指数和Simpson优势度指数显著低于单接种Xc或JW-JS1和CK(P<0.05);不同接种处理NL-895杨根际土壤微生物对BIOLOG微孔板中6大类碳源的利用能力差异显著(P<0.05)。【结论】荧光假单胞菌JW-JS1与外生菌根真菌Xc共接种对NL-895杨根际土壤酶活性和土壤微生物功能多样性的提高具有一定的促进作用,有利于其土壤肥力的改善,进而促进NL-895杨的生长。与单接种相比,溶磷细菌与外生菌根真菌共接种可更有效地提高杨树根际土壤酶活性和土壤微生物多样性。     

奶牛乳房炎大肠杆菌刃天青微量板快速诊断及药敏试验

为了建立奶牛乳房炎大肠杆菌快速诊断与药敏试验方法,用刃天青钠取代大肠杆菌选择培养基MQQ中的酚红指示剂,通过对刃天青的工作浓度、大肠杆菌接种量和培养时间等参数的研究,建立了奶牛乳房炎大肠杆菌刃天青微量板快速诊断方法.将不同细菌及其感染模拟奶样进行刃天青微量板快速培养结果显示,此刃天青微量板对大肠杆菌具有很强的选择培养特性,能在10 h内获得明确诊断结果,并能反映奶牛乳腺大肠杆菌感染强度.对89份临床奶样进行大肠杆菌刃天青微量板快速诊断敏感性和特异性检测,结果显示,此刃天青微量板法诊断大肠杆菌阳性奶样敏感性高达100%,诊断大肠杆菌阴性奶样特异性高达94.9%,总符合率高达95.5%.在此基础上用矫正浓度10种抗生素包被微量板,分别用10株大肠杆菌纯培养菌株及其感染模拟奶样和10份大肠杆菌阳性奶样进行快速药敏试验,所获结果与标准纸片扩散法完全一致.这些研究结果表明,本研究建立的刃天青微量板法可以取代常规方法用于奶样大肠杆菌的快速诊断及药敏试验.     

A Resazurin Microplate Assay for Quick Diagnosis and Antibiotic Sensitivity Measurement of Bovine Mastitis-causing E.coli

To establish a microplate assay for quick diagnosis and antibiotic sensitivity measurement of bovine mastitis-causing E.coli,the phenol red indicator in E.coli selective medium was replaced with resazurin and the resazurin microplate assay was established by determing of optimization of indicator concentration,bacterial inoculation dose and culture time.The testing results of different bacteria and their infection-mimicking milk samples showed that the microplate assay had a strong selective property for E.coli and the definite diagnostic result could be obtained within 10 h with a capability of reflecting infection strength of dairy cow mammary gland.The results of 89 mastitis-positive milk samples showed that the microplate assay had a detection sensitivity of 100%,and the negative milk samples specificity was 94.9%,95.5% agreement to the conventional culture-based method.The micoplates were coated with rectified concentrations of 10 different antibiotics and the quick drug sensitivity assay was performed using 10 E.coli strains,infection-mimicking milk samples and 10 E.coli-positive milk samples.The obtained results had a complete agreement with that of standard disc diffusion method.These data suggested that the resazurin microplate assay established in this study could replace the conventional methods for quick diagnosis and antibiotic sensitivity measurement of bovine mastitis-causing E.coli.     

用紫外-荧光微孔板酶检测技术测定两种土壤的酶活性

【目的】土壤酶活性是土壤质量的重要指示指标,本文对土壤酶的快速检测方法进行了探讨。【方法】采用荧光微孔板酶检测技术,测定土壤硫酸酯酶、磷酸酶、β-葡萄糖苷酶和肽酶活性,采用紫外微孔板酶检测技术测定多酚氧化酶和过氧化物酶活性。【结果】结果表明,采自农田的赤红壤的硫酸酯酶、磷酸酶和肽酶活性高于紫色土和采自矿山的赤红壤的相应的酶活性;矿山赤红壤的多酚氧化酶和过氧化物酶活性在三者中最高;紫色土的β-葡萄糖苷酶活性高于赤红壤的β-葡萄糖苷酶活性。【结论】紫外-荧光微孔板酶检测技术可快速测定土壤中酶的活性。     

Herbicidal action of 2-hydroxy-3-alkyl-1,4-naphthoquinones

The main mode of herbicidal activity of 2-hydroxy-3-alkyl-1,4-naphthoquinones is shown to be inhibition of photosystem II (PSII). The herbicidal and in vitro activities have been measured and correlated with their (Log)octanol/water partition coefficients (Log Ko/w). The length of the 3-n-alkyl substituent for optimal activity differed between herbicidal and in vitro activity. The maximum in vitro activity was given by the nonyl to dodecyl homologues (Log Ko/w between 6.54 and 8.12), whereas herbicidal activity peaked with the n-hexyl compound (Log Ko/w = 4.95). The effect of chain branching was also investigated using isomeric pentyl analogues substituted at position 3. All exhibited similar levels of in vitro activities but herbicidal activities differed, albeit moderately, with the exception of one analogue that was much less phytotoxic. Other modes of action were also investigated using two representative compounds. They did not show any activity on photosystem I or mitochondrial complex I, or generate toxic oxygen radicals by redox cycling reactions. Only moderate activity was found against mitochondrial complex III from plants, in contrast to much higher corresponding activity using an insect enzyme.     

基于智能手机的便携式多功能食品安全快检系统开发

为解决目前食品安全快速检测系统集成化程度低、图像采集装置便携性差的问题,以智能手机为检测工具,采用数字图像比色法,对莱克多巴胺的现场快速定量检测问题进行研究。设计便携式图像采集装置固定拍摄环境,使用菲涅尔透镜汇聚光线,利用智能手机准确获得检测卡与微孔板的图像信息;设计基于OpenCV的图像处理算法和智能手机应用程序,在移动终端实现对检测卡和微孔板反应区域的自动识别和颜色特征的分析。结果表明：1）本研究设计的图像采集装置可以对检测卡与微孔板准确成像,颜色特征变异系数CV<3%;2）利用免疫层析法和免疫吸附法对莱克多巴胺进行快速检测,2种方法对应的颜色特征值与莱克多巴胺的质量浓度均呈对数关系,决定系数R²>0.97,其中,免疫吸附法的检测结果与酶标仪检测结果基本一致（R²=0.987 3）。本系统具有便携、快速等优点,不需要实验室设备,适用于非专业人员在现场对莱克多巴胺进行快速定量检测。