A single extraction method for the analysis by liquid chromatography/tandem mass spectrometry of fumonisins and biomarkers of disrupted sphingolipid metabolism in tissues of maize seedlings
A single extraction method for the analysis by liquid chromatography/tandem mass spectrometry of fumonisins and biomarkers of disrupted sphingolipid metabolism in tissues of maize seedlings
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DOI：
10.1007/s00216-008-2166-x
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发表时间：
2008-07-01
影响因子：
4.3
通讯作者：
Riley, Ronald T.
中科院分区：
文献类型：
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作者：
Zitomer, Nicholas C.;Glenn, Anthony E.;Riley, Ronald T.
轮枝镰刀菌（Fusarium verticillioides）是许多植物的病原体，产生伏马菌素。除了对动物毒性的充分研究外，这些毒素还有助于易感玉米品种的玉米幼苗疾病的发展。伏马菌素破坏鞘脂生物合成发生在发病过程中。建立了玉米组织中伏马菌素B（1）（FB（1））、B（2）（FB（2））和B（3）（FB（3））、游离鞘氨醇碱和鞘氨醇碱1-磷酸盐的提取-液相色谱/线性离子阱串联质谱分析方法。该方法涉及使用1：1乙腈：水+ 5%甲酸（1 ml/10 mg组织）进行单次浸提。平均回收率范围约为50%至99%，检测限范围为10 fg μ l（-1）至6900 fg μ l（-1）。为了测试该方法的有效性，用致病性伏马菌素产生菌株接种敏感玉米品系的种子。轮枝状然后收获幼苗，并在叶和根组织中测量伏马菌素含量以及鞘氨醇碱及其1-磷酸盐。伏马菌素积累显着更大的叶1相比，叶2和3。FB（1）、FB（2）和FB（3）在根组织中检测到，FB（1）优先在叶组织中积累。鞘氨醇碱和它们的1-磷酸盐的积累在从接种的种子生长的幼苗的根和叶中是明显的，在叶1、2和3中积累的水平是相似的。所开发的方法是有效的，快速的，灵敏的，用于同时测量组织中的伏马菌素及其对疾病的鞘脂代谢物生物标志物的影响。该方法可用于玉米品种对镰刀菌的感病性筛选。轮枝孢菌引起的幼苗病害。
The fungus Fusarium verticillioides is a pathogen of many plants and produces fumonisins. In addition to their well-studied animal toxicoses, these toxins contribute to the development of maize seedling disease in susceptible maize varieties. Fumonisin disruption of sphingolipid biosynthesis occurs during pathogenesis. An extraction method was developed for the simultaneous analysis of fumonisins B(1) (FB(1)), B(2) (FB(2)) and B(3) (FB(3)), free sphingoid bases and sphingoid base 1-phosphates in maize tissues by liquid chromatography/linear ion trap tandem mass spectrometry. The method involved a single extraction using 1:1 acetonitrile:water + 5% formic acid (1 ml per 10 mg tissue). Mean recoveries ranged from approximately 50 to 99 percent, and limits of detection ranged from 10 fg mu l(-1) to 6900 fg mu l(-1). To test the efficacy of the method, seeds of a susceptible maize line were inoculated with a pathogenic, fumonisin-producing strain of F. verticillioides. The seedlings were then harvested, and fumonisin content, as well as sphingoid bases and their 1-phosphates, were measured in the leaf and root tissues. Fumonisin accumulation was significantly greater in leaf one compared to leaves two and three. While FB(1), FB(2), and FB(3) were detected in root tissues, FB(1) was preferentially accumulated in leaf tissues. Accumulation of sphingoid bases and their 1-phosphates was evident in roots and leaves of seedlings grown from inoculated seed, with the level of accumulation being similar in leaves 1, 2 and 3. The method developed was effective, fast, and sensitive for use in simultaneously measuring fumonisin in tissues and their effects on sphingolipid metabolite biomarkers of disease. The method should be useful for screening maize cultivars for susceptibility to F. verticillioides-induced seedling diseases.