Garibaldi A, Bertetti D, Gullino M L
Centre of Competence for the Innovation in the Agro-Environmental Sector (AGROINNOVA) Via Leonardo da Vinci 44, 10095 Grugliasco, Italy.
Plant Dis. 2009 Sep;93(9):969. doi: 10.1094/PDIS-93-9-0969B.
Platycodon grandiflorum (balloon flower), a perennial plant belonging to the Campanulaceae family, is widely grown as a bedding plant in temperate gardens. This species is characterized by the ability to bloom profusely throughout the summer into early fall and for its white to blue and pink flowers. In September 2008, symptoms of a previously unknown blight were observed in six gardens located in the Biella Province of northern Italy. When the disease developed, temperatures ranged between 15 and 22°C with frequent rains (149.8 mm of rainfall registered in September 2008 by the meteorological station of Oropa, located in the same area in which the disease appeared). Initially, leaves and petioles appeared chlorotic. Subsequently, lesions developed on the stems and flowers were sometimes affected. In each garden examined, approximately 50% of the plants were affected by the disease. A soft, gray mycelium was observed on symptomatic tissues, especially the stems. Severely infected leaves and stems eventually became completely rotted and later desiccated. Diseased tissue was excised from affected leaves, immersed in a solution containing 1% sodium hypochlorite for 10 s, and then cultured on potato dextrose agar (PDA) medium. A fungus developed that produced abundant mycelium on PDA medium when incubated under constant fluorescent light at 22 ± 1°C. Numerous sclerotia were produced on PDA plates incubated for 20 days at 8 ± 1°C. Sclerotia were dark, irregular, and measured 1 to 3.5 × 0.9 to 2.5 (average 2.1 × 1.5) mm. Conidia were smooth, ash colored, unicellular, ovoid, and measured 11 to 19 × 7 to 13 (average 15 × 11) μm. These morphological features were typical of those described for Botrytis cinerea (2). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 539-bp segment showed 100% similarity with the sequence of Botryotinia fuckeliana (perfect stage of B. cinerea). The nucleotide sequence has been assigned the GenBank Accession No. GQ149480. Pathogenicity tests were performed by placing 1-cm fragments removed from PDA cultures of B. cinerea isolated from balloon flower on leaves of healthy potted P. grandiflorum plants (4-month-old). Five fragments were placed on each plant. Plants inoculated with PDA alone served as controls. Ten plants per treatment were used. Plants were covered with plastic bags for 5 days after inoculation and maintained in a greenhouse at temperatures between 18 and 23°C. The first foliar lesions developed on leaves 3 days after inoculation, and after 5 days, 80% of the leaves were severely infected. As the infection progressed after the inoculation, the stems also became infected. Control plants remained healthy. B. cinerea was consistently reisolated from leaf and stem lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of B. cinerea on P. grandiflorum in Italy, as well as in Europe. Blight on balloon flower attributed to Botrytis spp. was previously reported in the United States (3). References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, England, 1971. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.
桔梗,一种属于桔梗科的多年生植物,在温带花园中广泛作为花坛植物种植。该物种的特点是整个夏季到初秋都能大量开花,花朵颜色从白色到蓝色和粉色。2008年9月,在意大利北部比耶拉省的六个花园中观察到一种此前未知的枯萎病症状。病害发生时,温度在15至22°C之间,降雨频繁(2008年9月位于病害出现区域的奥罗帕气象站记录降雨量为149.8毫米)。最初,叶片和叶柄出现褪绿现象。随后,茎上出现病斑,花朵有时也会受到影响。在每个检查的花园中,约50%的植株受到病害影响。在有症状的组织上,尤其是茎上,观察到柔软的灰色菌丝体。严重感染的叶片和茎最终完全腐烂,随后干枯。从受影响的叶片上切下病组织,浸入含1%次氯酸钠的溶液中10秒,然后在马铃薯葡萄糖琼脂(PDA)培养基上培养。培养出一种真菌,在22±1°C的恒定荧光灯下培养时,该真菌在PDA培养基上产生大量菌丝体。在8±1°C下培养20天的PDA平板上产生了大量菌核。菌核颜色深,形状不规则,大小为1至3.5×0.9至2.5(平均2.1×1.5)毫米。分生孢子光滑,灰白色,单细胞,卵形,大小为11至19×7至13(平均15×11)微米。这些形态特征与灰葡萄孢菌(2)描述的典型特征相符。使用引物ITS4/ITS6扩增rDNA的内部转录间隔区(ITS)并进行测序。对539碱基对片段的BLAST分析(1)显示与富克尔葡萄孢(灰葡萄孢的有性阶段)的序列100%相似。该核苷酸序列已被赋予GenBank登录号GQ149480。通过将从桔梗分离的灰葡萄孢的PDA培养物中切下的1厘米片段放置在健康盆栽桔梗植株(4个月大)的叶片上进行致病性测试。每株植物放置5个片段。仅接种PDA的植物作为对照。每个处理使用10株植物。接种后用塑料袋覆盖植物5天,并在18至23°C的温室中养护。接种后3天叶片上出现最初的叶斑,5天后,80%的叶片受到严重感染。随着接种后感染的进展,茎也被感染。对照植物保持健康。从叶片和茎的病斑中持续重新分离出灰葡萄孢菌。致病性测试完成了两次。据我们所知,这是意大利以及欧洲关于桔梗上存在灰葡萄孢菌的首次报道。此前在美国报道过桔梗上由葡萄孢属引起的枯萎病(3)。参考文献:(1)S.F.阿尔茨舒尔等人,《核酸研究》25:3389,1997年。(2)M.B.埃利斯,《暗色丝孢菌》,英国皇家植物园邱园真菌研究所,1971年。(3)D.F.法尔等人,《美国植物和植物产品上的真菌》,美国植物病理学会,明尼苏达州圣保罗,1989年。