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Cercosporin From Pseudocercosporella capsellae and its Critical Role in White Leaf Spot Development

机译：cap假单孢菌中的Cercosporin及其在白叶斑发育中的关键作用

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Pseudocercosporella capsellae, the causative agent of white leaf spot disease in Brassicaceae, can produce a purple-pink pigment on artificial media resembling, but not previously confirmed as, the toxin cercosporin. Chemical extraction with ethyl acetate from growing hyphae followed by quantitative (thin-layer chromatography [TLC] and high-performance liquid chromatography [HPLC]) and qualitative methods showed an identical absorption spectrum, with similar retardation factor (Rf) values on TLC papers and an identical peak with the same retention time in HPLC as for a standard for cercosporin. We believe this is the first report to confirm that the purple pink pigment produced by P. capsellae is cercosporin. Confocal microscopy detected green autofluorescence of cercosporin-producing hyphae, confirming the presence of cercosporin inside hyphae. The highly virulent UWA Wlra-7 isolate of P. capsellae produced the greatest quantity of cercosporin (10.69 mg g(-1)). The phytotoxicity and role of cercosporin in disease initiation across each of three Brassicaceae host species (Brassica juncea, B. napus, and Raphanus raphanistrum) was also studied. Culture filtrates containing cercosporin were phytotoxic to all three host plant species, producing large, white lesions on highly sensitive B. juncea, only water-soaked areas on least sensitive R. raphanistrum, and intermediate lesions on B. napus. It is noteworthy that sensitivity to cercosporin of these three host species was analogous to their susceptibility to the pathogen, viz., B. juncea the most susceptible, R. raphanistrum the least susceptible, and B. napus intermediate. The presence of cercosporin in the inoculum significantly increased disease severity on the highly cercosporin-sensitive B. juncea. We believe that this is the first study to demonstrate that P. capsellae produces cercosporin in liquid culture rather than agar media. Finally, this study highlights an important role of cercosporin as a pathogenicity factor in white leaf spot disease on Brassicaceae as evidenced by the ability of the cercosporin-rich culture filtrate to reproduce white leaf spot lesions on host plants and by the enhanced virulence of P. capsellae in the presence of cercosporin.
机译：芥菜假单胞菌（Pseudocercosporella capsellae）是十字花科白叶斑病的病原体，可以在人工培养基上产生紫色粉红色色素，类似于但未证实是毒素铜尾孢菌素。从生长的菌丝中用乙酸乙酯进行化学萃取，然后进行定量（薄层色谱[TLC]和高效液相色谱[HPLC]）和定性方法，其吸收光谱相同，TLC纸上的阻滞因子（Rf）值相近。与头孢菌素标准品相同的峰，在HPLC中的保留时间相同。我们相信这是第一个证实由卡氏假单胞菌产生的紫色粉红色色素是头孢菌素的报告。共聚焦显微镜检测到产生头孢菌素的菌丝的绿色自发荧光，证实菌丝内部存在头孢菌素。衣原假单胞菌的高毒性UWA Wlra-7分离株产生最大量的头孢菌素（10.69 mg g（-1））。还研究了十字花科在三种十字花科宿主物种（芸苔芥，油菜双歧杆菌和Raphanus raphanistrum）中的植物毒性和在疾病引发中的作用。含有尾孢菌素的培养物滤液对所有三种寄主植物均具有植物毒性，在高度敏感的芥菜油菜上产生大的白色病斑，在最不敏感的萝卜油菜上仅浸水，而在油菜中产生中等病斑。值得注意的是，这三种寄主物种对头孢菌素的敏感性类似于它们对病原体的敏感性，即最易感的芥菜芽胞杆菌，最不易感的R. raphanistrum和中间的B. napus。在高度对头孢菌素敏感的芥菜芽孢杆菌中，接种物中头孢菌素的存在显着增加了疾病的严重程度。我们认为，这是第一项证明衣藻毕赤酵母在液体培养中而不是在琼脂培养基中产生头孢菌素的研究。最后，这项研究突出了头孢菌素作为十字花科白叶斑病致病性因子的重要作用，这由富含头孢菌素的培养滤液在宿主植物上繁殖白叶斑病的能力以及增强的P毒力所证明。头孢菌素存在下的s菜。

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