Andolfi A, Cimmino A, Evidente A, Iannaccone M, Capparelli R, Mugnai L, Surico G
Dipartimento di Scienze del Suolo, della Pianta, dell'Ambiente e delle Produzioni Animali, Università di Napoli Federico II.
Facoltà di Scienze Biotecnologiche, Università di Napoli Federico II, Via Università 100, 80055 Portici, Italy.
Plant Dis. 2009 Jul;93(7):680-684. doi: 10.1094/PDIS-93-7-0680.
A flow cytometry technique that unequivocally identifies some of the toxic metabolites produced by Phaeomoniella chlamydospora, one of the main fungal pathogens causing esca disease of grapevine (Vitis vinifera), was developed. Antibodies raised against exopolysaccharides (EPS)-metabolites produced by Pa. chlamydospora that have been reported to be phytotoxic-were used as antigen to immunize rats. The specificity of these antibodies was assayed by flow cytometry against Pa. chlamydospora polysaccharides and against EPS with a different structure isolated from other phytopathogenic fungi, including Phaeoacremonium aleophilum and the Botryosphaeriaceae species Neofusicoccum luteum and N. parvum. Using this method, Pa. chlamydospora polysaccharides were detected in the symptomatic leaves of esca-affected grapevines, while healthy and asymptomatic leaves from both healthy and diseased vines did not produce a binding reaction. This method potentially could be used to develop a simple kit to study the mechanisms underlying the development of esca foliar symptoms and to indirectly assess the presence of Pa. chlamydospora in grapevine material.
一种流式细胞术被开发出来，它能明确识别由葡萄座腔菌（Phaeomoniella chlamydospora）产生的一些有毒代谢产物，葡萄座腔菌是导致葡萄（Vitis vinifera）埃斯卡病的主要真菌病原体之一。针对葡萄座腔菌产生的胞外多糖（EPS）代谢产物（据报道具有植物毒性）制备的抗体被用作抗原免疫大鼠。通过流式细胞术检测这些抗体对葡萄座腔菌多糖以及从其他植物病原真菌（包括嗜土栖色串孢菌（Phaeoacremonium aleophilum）和葡萄座腔菌科物种新壳梭孢菌（Neofusicoccum luteum）和细小新壳梭孢菌（N. parvum））分离出的具有不同结构的EPS的特异性。使用这种方法，在受埃斯卡病影响的葡萄树有症状的叶片中检测到了葡萄座腔菌多糖，而来自健康和患病葡萄树的健康无症状叶片均未产生结合反应。该方法有可能用于开发一种简单的试剂盒，以研究埃斯卡叶部症状发展的潜在机制，并间接评估葡萄材料中葡萄座腔菌的存在情况。
