McGrath M T, Strauss J, Dillard H R
Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Long Island Horticultural Research and Extension Center, Riverhead, NY 11901.
Department of Plant Pathology and Plant-Microbe Biology, Cornell University, New York State Agricultural Experiment Station, Geneva 14456.
Plant Dis. 2011 Aug;95(8):1028. doi: 10.1094/PDIS-01-11-0040.
Phytophthora capsici Leonian is an important pathogen of solanaceous and cucurbit crops. Phytophthora blight was first reported on snap bean (Phaseolus vulgaris L.) in Michigan in 2003 (2) and Connecticut in 2010 (3). This report documents the discovery of P. capsici on snap bean (cv. Bronco) grown in Riverhead, NY in September 2008 and August 2010 on snap bean (cv. Valentino) in Holley, NY, more than 690 km away. Disease was favored by frequent rainfall and prolonged wet periods with air temperatures of 24 to 29°C. Both locations were commercial fields previously planted to pepper or cucurbits affected by P. capsici. In Riverhead, infected pods had characteristic yeast-like growth of P. capsici, which were predominantly sporangia. In Holley, large water-soaked lesions were observed on snap bean foliage, and as the disease progressed, leaves became necrotic and detached from the plant. Reddish brown lesions were observed on stems in advance of white areas of sporulation. Infected pods displayed white mycelial growth, were shriveled, and desiccated. P. capsici was isolated from symptomatic tissues. Stems and pods were surface disinfested for 3 min in 0.525% sodium hypochlorite solution, rinsed for 3 min in sterile distilled water, transferred to PARPH (4) media, and incubated at 22°C. After 5 days, hyphae from colony margins were excised and transferred to 15% unclarified V8 agar media. Cultures consisting of white mycelia and ovoid papillate sporangia on long pedicels were identified as P. capsici. Sporangia were 25.0 to 70.0 × 10.0 to 22.5 μm (average 42.0 × 16.25 μm). Identity was further confirmed by PCR primers specific to P. capsici (1). DNA was extracted from mycelia produced on V8 agar and amplification with the species-specific primers resulted in a PCR product of the same size as that obtained from a known isolate of P. capsici. Pathogenicity of the isolate from Holley was determined by two methods on 50-day-old snap bean plants (cv. Valentino) grown in a greenhouse. In method one, four plants were inoculated with 1-cm-diameter mycelial plugs excised from 8-day-old cultures. A single plug was placed against the stem at the soil line. Four control plants were treated similarly with noncolonized agar plugs. In method two, entire plants were atomized with 10 ml of a zoospore suspension (2.6 × 10/ml). Control plants were atomized with sterile distilled water. All plants were placed in a growth chamber with continuous mist for 24 h at 24°C. After 24 h, plants were enclosed in plastic bags and placed in a greenhouse at 27°C. Stem lesions similar to those observed in affected fields were evident on plants treated with mycelia plugs 2 days after inoculation. Plants inoculated with the zoospore suspension developed stem lesions and desiccated pods. Control plants were asymptomatic. P. capsici was successfully recovered from infected plant tissue, fulfilling Koch's postulates. The Riverhead isolate was demonstrated as pathogenic on snap bean and cucumber by placing colonized plugs on pods and fruit that were subsequently incubated in moist chambers (24°C, 90 to 100% relative humidity). P. capsici was successfully recovered from symptomatic pods and fruit. To our knowledge, this is the first report of Phytophthora blight caused by P. capsici on snap bean in New York. References: (1) A. R. Dunn et al. Plant Dis. 94:1461, 2010. (2) A. J. Gevens et al. Plant Dis. 92:201, 2008. (3) J. A. LaMondia et al. Plant Dis. 94:134, 2010. (4) G. C. Papavisas et al. Phytopathology 71:129, 1981.
辣椒疫霉(Phytophthora capsici Leonian)是茄科和葫芦科作物的一种重要病原菌。2003年在密歇根州(2)以及2010年在康涅狄格州(3)首次报道了菜豆(Phaseolus vulgaris L.)上发生的疫病。本报告记录了2008年9月在纽约州里弗黑德种植的菜豆(品种Bronco)上以及2010年8月在距离里弗黑德690多公里外的纽约州霍利种植的菜豆(品种Valentino)上发现辣椒疫霉的情况。频繁降雨以及气温在24至29°C的长时间潮湿天气有利于病害发生。这两个地点均为之前种植过受辣椒疫霉影响的辣椒或葫芦科作物的商业田地。在里弗黑德,受感染的豆荚上有辣椒疫霉典型的酵母样生长物,主要是孢子囊。在霍利,菜豆叶片上观察到大片水渍状病斑,随着病害发展,叶片坏死并从植株上脱落。在出现白色产孢区域之前,茎上观察到红褐色病斑。受感染的豆荚呈现白色菌丝生长,皱缩且干枯。从有症状的组织中分离出了辣椒疫霉。将茎和豆荚在0.525%次氯酸钠溶液中表面消毒3分钟,在无菌蒸馏水中冲洗3分钟,转移至PARPH(4)培养基上,并在22°C下培养。5天后,从菌落边缘切下菌丝转移至15%未澄清的V8琼脂培养基上。由白色菌丝体和长梗上的卵形乳头状孢子囊组成的培养物被鉴定为辣椒疫霉。孢子囊大小为25.0至70.0×10.0至22.5μm(平均42.0×16.25μm)。通过辣椒疫霉特异性PCR引物(1)进一步确认了其身份。从V8琼脂上生长的菌丝体中提取DNA,用种特异性引物扩增后得到的PCR产物大小与从已知辣椒疫霉分离物中获得的相同。通过两种方法在温室中种植的50日龄菜豆(品种Valentino)植株上测定了来自霍利的分离物的致病性。方法一,用从8日龄培养物上切下的直径1厘米的菌丝块接种4株植株。将单个菌丝块贴放在茎基部贴近土壤处。4株对照植株用未定植的琼脂块进行类似处理。方法二,用10毫升游动孢子悬浮液(2.6×10/ml)对整株植株进行喷雾。对照植株用无菌蒸馏水喷雾。所有植株置于生长室中,在24°C下持续喷雾24小时。24小时后,将植株装入塑料袋并置于27°C的温室中。接种菌丝块处理的植株在接种2天后茎上出现了与在受感染田地中观察到的类似病斑。接种游动孢子悬浮液的植株出现茎部病斑和干枯的豆荚。对照植株无症状。从受感染的植物组织中成功重新分离出辣椒疫霉,满足了柯赫氏法则。通过将定植的菌丝块放在豆荚和果实上,随后在保湿箱(24°C,相对湿度90至100%)中培养,证明了里弗黑德分离物对菜豆和黄瓜具有致病性。从有症状的豆荚和果实中成功重新分离出辣椒疫霉。据我们所知,这是纽约州菜豆上由辣椒疫霉引起的疫病的首次报道。参考文献:(1)A. R. Dunn等人,《植物病害》94:1461,2010。(2)A. J. Gevens等人,《植物病害》92:201,2008。(3)J. A. LaMondia等人,《植物病害》94:134,2010。(4)G. C. Papavisas等人,《植物病理学》71:129,1981。