应用高效离子交换色谱分析青枯菌的致病力分化  

Analysis of Different Virulent Ralstonia solanacearum Strains Using High Performance Ion Exchange Chromatography

作　　者：林娟[1] 刘树滔[1] 高珍娜[1] 饶平凡[1] 

机构地区：[1]福州大学生物科学与工程学院,福州350002

出　　处：《应用与环境生物学报》2009年第5期713-718,共6页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(No.20475010);福建省自然科学基金项目(No.2008J0066);福建省高等学校新世纪优秀人才支持计划项目(No.XSJRC2007-09);福州大学人才基金项目资助~~

摘　　要：应用高效离子交换色谱和激光光散射仪检测器对不同致病力的青枯菌进行分析,建立了一种快速检测青枯菌致病力分化的新方法.青枯菌纯培养物经过高效离子交换色谱分离得到3个致病力不同的特征峰,大小依次为峰3组分>峰2组分>峰1组分.对10株青枯菌进行色谱分析,并结合番茄组培苗感染试验检测其致病力,结果发现,强致病力菌株经过色谱分离只在峰3的保留时间位置出现单一特征峰,在9d内即可引起100%的番茄组培苗发病;若菌株经过色谱分离形成3个特征峰,则峰3所占的面积比越大,该菌株的致病性相对就越强.25株不同致病力青枯菌的验证试验表明了该方法的可行性,番茄组培苗发病率x与峰3面积比y之间呈现出良好的线性关系,回归方程y=0.9581x+5.4984,相关系数r=0.986.通过对青枯菌色谱行为、致病力、细胞表面黏附的EPSⅠ含量三者之间相互关系的进一步研究,发现青枯菌的致病力越强,则细胞表面黏附的EPSⅠ越多,峰3所占的面积比就越大.High performance ion exchange chromatography （HPLC） coupled with laser light scattering instrument was employed for analyzing different virulent Ralstonia solanacearum strains, and a new method for rapidly detecting the pathogenicity of R. solanacearum was established. The pure culture of R. solanacearum was successfully separated into three characteristic fractions of different virulences by HPLC. The virulence of peak 3 fraction was the strongest and that of peak 1 fraction the weakest. Ten strains were analyzed by HPLC and applied to infect tomato tissue culture plantlets using leaf-cutting method to further determine their virulence. The results showed that strong virulent strains formed only one characteristic peak at the retention time of peak 3, which could make 100% of tissue culture plantlets die within nine days. If the strains were separated into three characteristic peaks, the area ratio of peak 3 was increasing with the virulence of R. solanacearum. Furthermore, twenty-five R. solanacearum strains with different virulent activities were analyzed by HPLC to validate the feasibility of this method. The correlation between infection mortality of tomato tissue culture plantlets （x） and peak 3 area ratio （y） was linear, with the regression equation ofy = 0.9581x ＋ 5.4984, and correlation coefficient r = 0.986. The relationships among chromatographic behaviors, virulence and EPSI contents of cell surface were further investigated and a positive correlation was observed among them. Fig 3, Tab 6, Ref 15

关 键 词：青枯菌 致病力 高效离子交换色谱 激光光散射仪 

分 类 号：S432.42[农业科学—植物病理学]