Gao L, Feng C, Li B, Liu T, Liu B, Chen W
State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences (CAAS), Beijing, China.
J Appl Microbiol. 2015 Feb;118(2):497-505. doi: 10.1111/jam.12703. Epub 2014 Dec 21.
Tilletia controversa is an internationally quarantined pathogenic fungus that causes dwarf bunt of wheat and is similar to Tilletia caries in both teliospore morphology and genetic structure. This study developed a rapid and sensitive immunofluorescence method for differentiating the teliospores of T. controversa from T. caries.
The method utilizes monoclonal antibody D-1 against teliospores of T. controversa as well as a PE-Cy3-conjugated goat anti-mouse antibody (overlapping light excitation of 495 and 555 nm). The orange cycle fluorescent signal was stronger against T. controversa teliospores in the outer spore wall and net ridge, whereas only the green signal was observed for the protoplasm of T. caries teliospores. The detection limit of this method was 2.0 μg ml(-1) of the D-1 monoclonal antibody.
This study describes the production and diagnostic application of a novel mouse monoclonal antibody specific to T. controversa teliospores.
This method could be used for the on-site identification of T. controversa teliospores in the near future and will help in selecting fungicides to control dwarf bunt of wheat as further technical developments are achieved.
小麦矮腥黑穗病菌是一种国际检疫性致病真菌，可引起小麦矮腥黑穗病，其冬孢子形态和遗传结构与小麦网腥黑穗病菌相似。本研究开发了一种快速灵敏的免疫荧光方法，用于区分小麦矮腥黑穗病菌和小麦网腥黑穗病菌的冬孢子。
该方法利用抗小麦矮腥黑穗病菌冬孢子的单克隆抗体D-1以及PE-Cy3偶联的山羊抗小鼠抗体（495和555nm重叠光激发）。橙色循环荧光信号在小麦矮腥黑穗病菌冬孢子的外孢壁和网脊处更强，而小麦网腥黑穗病菌冬孢子的原生质仅观察到绿色信号。该方法的检测限为2.0μg ml(-1)的D-1单克隆抗体。
本研究描述了一种针对小麦矮腥黑穗病菌冬孢子的新型小鼠单克隆抗体的制备及其诊断应用。
该方法在不久的将来可用于小麦矮腥黑穗病菌冬孢子的现场鉴定，并将有助于在取得进一步技术进展时选择杀菌剂来防治小麦矮腥黑穗病。
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