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小鼠肠道菌群HPLC分析中色谱分离条件的优化
  • ISSN号:1671-8135
  • 期刊名称:《中国生物工程杂志》
  • 时间:0
  • 分类:O658[理学—分析化学;理学—化学]
  • 作者机构:[1]福州大学生物工程研究所,福建福州350002
  • 相关基金:国家自然科学基金资助项目(No.20475010)、福建省自然科学基金资助项目(No.Z0516003)和福州大学科技发展基金资助项目(No.2004-XQ-15).致谢:感谢福建省农业科学院生物技术研究所刘波博士和周先治博士为本研究提供的大力支持.
中文摘要:

应用高效离子交换色谱和激光光散射仪在线检测,快速分离定量不同致病力的青枯菌。青枯菌经过高效离子交换色谱分离得到3个特征峰,通过2,3,5-三苯基氯化四氮唑(TTC)平板鉴定和采用剪叶法回接番茄组培苗感染试验,发现这3个色谱峰所对应的青枯菌在致病力方面存在差异;其中峰3组分的致病力最强,峰1组分的致病力最弱。通过对青枯菌进样量与激光光散射仪的响应信号(峰面积)之间的线性关系研究,发现当青枯菌进样菌数为9×10^6~9×10^8时,菌数与色谱峰面积之间呈现出良好的线性关系,相关系数r=0.99。该项应用研究为不同致病力青枯菌的快速定量提供了一种新的分析方法。

英文摘要:

High performance ion exchange chromatography coupled with laser light scattering instrument was employed for the rapid separation and quantitation of Ralstonia solanacearum of different virulence. The pure culture of Ralstonia solanacearum was successfully separated into three characteristic fractions. Each fraction was collected and inoculated onto 2,3,5-triphenyltetrazolium chloride (TTC) plates to identify its virulence. The shapes and colors of the colonies were imaged, and the average attenuation index ( attenuation index = red spot diameter of colony/total colony diameter) of ten colonies of each fraction was carefully determined. Furthermore, each fraction was inoculated into SPA liquid media at 30℃ with shaking (200 r/min) for 48 h, the cells were harvested, suspended at a density of 1.2 × 10^9 cfu/mL, and applied to infect tomato tissue culture plantlets using leaf-cutting method. The infection mortality of the tomato tissue culture plantlets was recorded from 1 to 9 days after inoculation. The results showed that the virulences of each fraction were different on the basis of attenuation index and infection mortality. The virulence of peak 3 fraction was the strongest. On the contrary, the virulence of peak 1 fraction was the weakest. In addition, the linear relationships between different injection volumes ( 1 - 180 μL) and their peak areas were investigated. The linearity was good within the range of the bacterial number of 9 × 10^6 - 9 × 10^8 ( r = 0.99). This method can be potentially used as a novel tool for the rapid separation and quantitation of Ralstonia solanacearum of different virulences.

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期刊信息
  • 《中国生物工程杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中国科学院文献情报中心 中国生物技术发展中心 中国生物工程学会
  • 主编:张树庸
  • 地址:北京市中关村北四环西路33号
  • 邮编:100080
  • 邮箱:biotech@mail.las.ac.cn
  • 电话:010-82624544 82626611-6631
  • 国际标准刊号:ISSN:1671-8135
  • 国内统一刊号:ISSN:11-4816/Q
  • 邮发代号:82-673
  • 获奖情况:
  • 1991年中国科学院科技进步三等奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:12959