Montecchio L
Territorio e Sistemi Agro Forestali Department, University of Padova, via dell'Università 16, I-35020 Legnaro, Italy.
Plant Dis. 2005 Sep;89(9):1014. doi: 10.1094/PD-89-1014A.
During May of 2004, damping-off of an estimated 70% of beech (Fagus sylvatica L.) seedlings was observed in a bare-root forest nursery located in northern Italy. Twenty-eight days after sowing, cotyledonary leaves were chlorotic, wilted, and occasionally desiccated; stem and collars appeared stunted and discolored with yellowish gray-to-brownish longitudinal streaks arising from the foot region, and rootlets were partially to completely rotten. No fruiting bodies were present on or near the damaged regions. Seedling establishment was poor within the disease foci that gradually increased in size. Longitudinal sections through the damaged stems showed dark brown streaks in the vascular tissue, and microscopic examination revealed that vessels frequently contained mycelium. Ten symptomatic plants were selected, and isolations were made from the necrotic margins of stems previously surface sterilized with 1% sodium hypochlorite and thoroughly rinsed, longitudinally cut into two parts 5-mm long, placed on potato dextrose agar (PDA), and incubated at 22 ± 1°C for 5 days in the dark. Although a variety of microorganisms were isolated, Fusarium avenaceum (Fr.:Fr.) Sacc. (1) was always recovered. Artificial inoculations with the fungus were made on 20-day-old, container-grown beech seedlings. The epidermis surrounding the collar was surface sterilized with 1% sodium hypochlorite, rinsed, and gently scraped with a sterile scalpel. After masking the rest of the plant with a plastic sheet, the wound was sprayed with a conidial suspension containing 10 macroconidia per cm in water and sealed with Parafilm. Controls were treated the same way but with sterile water. Each treatment was applied to 10 seedlings and incubated in the greenhouse (20 ± 2°C, 80% relative humidity, and 12 h of natural light per day). After 20 days, wounds treated with F. avenaceum showed necrotic lesions that developed into small patches of dead epidermis. Radial sections through the stem 2 cm above the inoculation site from five plants showed the presence of mycelium in the vessels, from which the fungus was reisolated. Thirty days postinoculation, the remaining five plants showed the same symptoms observed in the nursery, and microscopic observations confirmed the presence of the fungus. No disease symptoms or mycelium were observed in the inner tissues of control plants. The pathogenicity test was repeated twice with the same results. The fungus was not detected by culturing 100 surface-sterilized seeds from the same stock that had been sown in the nursery. F. avenaceum has a broad host range including angiosperms and gymnosperms, and the described symptoms are fairly typical (2). To our knowledge, this is the first report of this disease in Italy. Further research on fungal survivability in nursery soil and plant debris is in progress. The isolate is preserved in the Centraalbureau voor Schimmelcultures (NL) collection as no. 115957. References: (1) C. Booth. Fusarium. Commonwealth Mycological Institute, Kew, UK, 1977. (2) H. Butin. Tree Diseases and Disorders. Oxford University Press, New York, 1995.
2004年5月期间,在意大利北部一家裸根苗森林苗圃中,估计有70%的山毛榉(欧洲山毛榉)幼苗出现猝倒病症状。播种28天后,子叶变黄、枯萎,偶尔干枯;茎和茎基部发育不良、变色,从基部向上出现黄灰色至褐色的纵向条纹,细根部分或全部腐烂。受损区域及其附近未发现子实体。病斑内的幼苗定植情况很差,且病斑面积逐渐扩大。对受损茎进行纵切,可见维管组织中有深褐色条纹,显微镜检查显示导管中常有菌丝体。挑选了10株有症状的植株,从先前用1%次氯酸钠进行表面消毒并彻底冲洗过的茎的坏死边缘进行分离,将茎纵向切成5毫米长的两部分,置于马铃薯葡萄糖琼脂(PDA)上,在22±1°C黑暗条件下培养5天。虽然分离出了多种微生物,但总是能分离到燕麦镰刀菌(Fusarium avenaceum (Fr.:Fr.) Sacc.)。用该真菌对20日龄的容器培育山毛榉幼苗进行人工接种。用1%次氯酸钠对茎基部周围的表皮进行表面消毒,冲洗后用无菌手术刀轻轻刮擦。用塑料薄膜覆盖植株的其余部分后,用含有每厘米10个大分生孢子的分生孢子悬浮液喷洒伤口,并用石蜡膜密封。对照组用无菌水进行相同处理。每种处理应用于10株幼苗,并在温室中培养(20±2°C,相对湿度80%,每天12小时自然光照)。20天后,用燕麦镰刀菌处理的伤口出现坏死病斑,发展成小块死亡表皮。对5株接种部位上方2厘米处的茎进行径向切片,显示导管中有菌丝体,并从中再次分离出该真菌。接种30天后,其余5株植株出现了在苗圃中观察到的相同症状,显微镜观察证实了真菌的存在。对照植株的内部组织未观察到病害症状或菌丝体。致病性试验重复了两次,结果相同。从苗圃中播种的同一批种子中选取100颗进行表面消毒后培养,未检测到该真菌。燕麦镰刀菌寄主范围广泛,包括被子植物和裸子植物,所描述的症状相当典型。据我们所知,这是该病在意大利的首次报道。目前正在对苗圃土壤和植物残体中的真菌存活能力进行进一步研究。该分离菌株保藏于荷兰中央真菌保藏中心(Centraalbureau voor Schimmelcultures),编号为115957。参考文献:(1) C. Booth. Fusarium. 英国皇家植物园邱园真菌研究所,1977年。(2) H. Butin. Tree Diseases and Disorders. 牛津大学出版社,纽约,1995年。