Yun H K, Louime C, Lu J
Center for Viticulture and Small Fruit Research, Florida A&M University, Tallahassee 32317.
Plant Dis. 2007 Jul;91(7):905. doi: 10.1094/PDIS-91-7-0905B.
Anthracnose of grapes is an economically devastating disease caused by Elsinoe ampelina (2). Warm, humid weather favors disease development, and therefore in the United States, it is generally restricted to grape-growing areas east of the Rocky Mountains. Vitis vinifera is highly susceptible to the disease, which is one of the principal factors preventing the development of an industry with this high-quality grape in the southeastern United States. Growers in this area produce local species-such as muscadine grapes (V. rotundifolia Michx.) and hybrids. Muscadine grapes are known for their resistance or "immunity" to many diseases found in bunch (Euvitis spp. Planch.) grape species (1). As yet, there has been no formal report of anthracnose or its causal agent on muscadine grapes. E. ampelina was detected on muscadine leaves for the first time in the experimental vineyard at the Center for Viticulture and Small Fruit Research during the summer of 2006. Approximately 40% of the 52 muscadine cultivars in the collection showed circular or irregular black spots typical of anthracnose mainly on young leaves and tendrils. However, no symptoms were observed on fruits, shoot tips, or any other plant part. To confirm the causal agent, infected leaves were surface sterilized with 75% ethanol, dipped in 2% sodium hypochlorite for 15 s, rinsed in distilled water, dissected into small 0.5-cm leaf discs, and plated on potato dextrose agar (PDA) and incubated at 28°C. Single-spore isolates were grown on PDA. Colonies were slow growing and appeared as dark red mounds with some mycelia. Conidia were cylindrical and hyaline with pointed ends consistent with previous reports for E. ampelina (2). The identity was also confirmed by using the following PCR primers to the 18S RNA: left primer; TCCGTAGGTGAACCTGCGGA and right primer; TCCTACCTGAT CCGAGGTCA designed on the basis of the alignment of E. ampelina sequences deposited in NCBI database. To fulfill Koch's postulates, symptoms were reproduced by artificial inoculation onto young muscadines (cv. Carlos) and bunch (cv. Cabernet Sauvignon) grapevines. A conidial suspension was prepared from single-conidial cultures, and three experimental vines of each species were sprayed with 0.5 ml of suspension (2 × 10 conidia per ml), whereas three control plants were sprayed with distilled water. The plants were incubated in a moist chamber at 28°C with 16 h of light. The first typical symptoms appeared on V. vinifera 4 days postinoculation and on the muscadines 6 days postinoculation. To our knowledge, this is the first report confirming anthracnose disease on muscadine grapes. References: (1) J. Lu et al. Acta Hortic. 528:479, 2000. (2) R. C. Pearson and A. C. Gohen. Anthracnose. Pages 18-19 in: Compendium of Grape Diseases. The American Phytopathological Society. St. Paul, MN, 1994.
葡萄炭疽病是由葡萄痂圆孢菌(Elsinoe ampelina)引起的一种具有经济毁灭性的病害(2)。温暖潮湿的天气有利于病害发展,因此在美国,该病通常局限于落基山脉以东的葡萄种植区。欧亚种葡萄(Vitis vinifera)对这种病害高度敏感,这是美国东南部阻碍利用这种优质葡萄发展产业的主要因素之一。该地区的种植者种植当地品种,如圆叶葡萄(V. rotundifolia Michx.)及其杂交品种。圆叶葡萄以对许多在欧亚种葡萄中发现的病害具有抗性或“免疫性”而闻名(1)。到目前为止,尚未有关于圆叶葡萄炭疽病或其病原菌的正式报道。2006年夏季,在葡萄栽培与小果研究中心的实验葡萄园首次在圆叶葡萄叶片上检测到葡萄痂圆孢菌。收集的52个圆叶葡萄品种中约40%的植株在幼叶和卷须上出现了典型的炭疽病圆形或不规则黑色斑点。然而,在果实、新梢顶端或任何其他植物部位未观察到症状。为了确定病原菌,将感染的叶片用75%乙醇进行表面消毒,浸入2%次氯酸钠中15秒,用蒸馏水冲洗,切成0.5厘米的小叶盘,接种到马铃薯葡萄糖琼脂(PDA)上,在28°C下培养。单孢分离株在PDA上生长。菌落生长缓慢,呈暗红色菌团,有一些菌丝体。分生孢子呈圆柱形,无色透明,两端尖锐,与先前关于葡萄痂圆孢菌的报道一致(2)。通过使用基于NCBI数据库中葡萄痂圆孢菌序列比对设计的以下18S RNA的PCR引物也证实了其身份:左引物;TCCGTAGGTGAACCTGCGGA,右引物;TCCTACCTGAT CCGAGGTCA。为了验证科赫法则,通过人工接种到幼龄圆叶葡萄(品种:卡洛斯)和欧亚种葡萄(品种:赤霞珠)葡萄植株上再现了症状。从单孢培养物中制备分生孢子悬浮液,每个品种的三株实验葡萄植株喷洒0.5毫升悬浮液(每毫升含2×10个分生孢子),而三株对照植株喷洒蒸馏水。将植株置于28°C、光照16小时的潮湿箱中培养。接种后4天在欧亚种葡萄上出现了第一批典型症状,接种后6天在圆叶葡萄上出现症状。据我们所知,这是首次证实圆叶葡萄发生炭疽病的报道。参考文献:(1)J. Lu等人,《园艺学报》528:479,2000。(2)R. C. Pearson和A. C. Gohen,《炭疽病》,载于《葡萄病害简编》,美国植物病理学会,明尼苏达州圣保罗,1994年,第18 - 19页。