Testen A L, McKemy J M, Backman P A
Department of Plant Pathology, The Pennsylvania State University, University Park, 16802.
USDA-APHIS-PPQ-National Identification Services, Beltsville, MD 20705.
Plant Dis. 2012 Jan;96(1):146. doi: 10.1094/PDIS-09-11-0728.
Quinoa, Chenopodium quinoa Willd., is an Andean crop prized for its high nutritional value and adaptability to harsh environments. Quinoa is plagued by downy mildew caused by Peronospora variabilis Gäum (formerly Peronospora farinosa f. sp. chenopodii Byford) (1). Quinoa production has spread beyond native Andean ranges and quinoa downy mildew has been reported in India, Canada, and Denmark (1). During the summer of 2011, quinoa trials were established to determine the ability of quinoa to grow under Mid-Atlantic conditions and monitor for regional disease problems. In July, after cool, rainy conditions, downy mildew-like symptoms were observed on quinoa at research plots in Centre and Lancaster counties of Pennsylvania. Symptoms and signs consisted of irregularly shaped areas of foliar chlorosis or pink discoloration accompanied by dense, gray sporulation on both leaf surfaces. Sporangia were tan to gray-brown, semi-ovoid, often with a pedicel, mean length of 31 μm, and mean width of 23 μm. Sporangiophores branched dichotomously, and the terminal branchlets curved and tapered to a point. Orange oospores were present in field samples of leaf tissue. DNA was extracted from infected foliar tissue and sporangial suspensions. A seminested PCR protocol (2) was used to obtain partial internal transcribed spacer (ITS) sequences of six Peronospora isolates. The sequences shared 99% maximum identity to a known P. variabilis accession (FM863721.2) in GenBank. A voucher specimen was deposited into the U.S. National Fungus Collections (BPI 882064). Pathogenicity of each of two strains of P. variabilis was confirmed by inoculating quinoa with sporangia (4). Sporangia were shaken from leaves in sterile distilled water and the suspension was filtered through cheesecloth. A 0.01% Tween solution was added and the suspension diluted to 10 sporangia/ml. With an atomizer, a 10-ml sporangial suspension (or sterile water for noninoculated control plants) was sprayed onto one flat of 18 2-week-old quinoa plants, and relative humidity was increased to saturation using a humidity dome for 24 h. After 1 week, chlorosis and pink discoloration were noted on leaves of inoculated quinoa, and after 18 h of subsequent increased humidity (>95% relative humidity), dense gray sporulation was observed. No symptoms were noted on noninoculated control plants. Sporangia and sporangiophores were examined morphologically and confirmed to be P. variabilis, confirming Koch's postulates. For culture maintenance, 2-week-old quinoa leaves were placed onto a sporangial suspension on top of 1% water agar and maintained in a growth chamber at 20°C with 16 h of light per day. Quinoa downy mildew is seedborne (3) and initial infections may have occurred from oospores in the pericarp, despite intensive processing of consumable quinoa seeds to remove saponins. To our knowledge, this is the first report of quinoa downy mildew in the United States and also the first report of P. variabilis in the United States. References: (1) Y. Choi et al. Mycopathologia 169:403, 2010. (2) D. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) S. Danielson et al. Seed Sci. Technol. 32:91, 2004. (4) J. Ochoa et al. Plant Pathol. 48:425, 1999.
藜麦(Chenopodium quinoa Willd.)是一种原产于安第斯山脉的作物,因其高营养价值和对恶劣环境的适应性而备受珍视。藜麦受到可变霜霉(Peronospora variabilis Gäum,原称粉霜霉藜亚种Peronospora farinosa f. sp. chenopodii Byford)引起的霜霉病困扰(1)。藜麦种植已扩展至安第斯山脉以外的地区,印度、加拿大和丹麦均有藜麦霜霉病的报道(1)。2011年夏季,开展了藜麦试验,以确定藜麦在大西洋中部地区的生长能力,并监测区域病害问题。7月,在凉爽多雨的天气过后,宾夕法尼亚州中心县和兰开斯特县的研究地块上,藜麦出现了类似霜霉病的症状。症状和病征包括叶片不规则形状的褪绿区域或粉红色变色,同时叶片两面出现密集的灰色霉层。孢子囊呈棕褐色至灰褐色,半卵形,通常有柄,平均长度为31μm,平均宽度为23μm。孢囊梗二叉状分枝,末端小枝弯曲并逐渐变细至一点。在叶片组织的田间样本中发现了橙色的卵孢子。从受感染的叶片组织和孢子囊悬液中提取DNA。采用半巢式PCR方案(2)获得了6个霜霉分离株的部分内部转录间隔区(ITS)序列。这些序列与GenBank中已知的可变霜霉登录号(FM863721.2)的最大同源性为99%。一份凭证标本已存入美国国家真菌收藏库(BPI 882064)。通过用孢子囊接种藜麦,证实了两株可变霜霉的致病性(4)。将孢子囊从叶片上摇落到无菌蒸馏水中,悬液经粗棉布过滤。加入0.01%的吐温溶液,将悬液稀释至每毫升含10个孢子囊。用喷雾器将10毫升孢子囊悬液(或用于未接种对照植株的无菌水)喷到一盆18株2周龄的藜麦植株上,使用湿度罩将相对湿度提高到饱和状态并保持24小时。1周后,接种藜麦的叶片出现褪绿和粉红色变色,随后湿度增加(相对湿度>95%)18小时后,观察到密集的灰色霉层。未接种的对照植株未出现症状。对孢子囊和孢囊梗进行形态学检查,确认是可变霜霉,从而证实了柯赫氏法则。为了进行菌种保存,将2周龄的藜麦叶片放在1%水琼脂上的孢子囊悬液上,置于生长室中,温度为20°C,每天光照16小时。藜麦霜霉病是种传病害(3),尽管对食用藜麦种子进行了强化加工以去除皂苷,但最初的感染可能源于果皮中的卵孢子。据我们所知,这是美国关于藜麦霜霉病的首次报道,也是美国关于可变霜霉的首次报道。参考文献:(1)Y. Choi等人,《真菌病理学》169:403,2010年。(2)D. Cooke等人,《真菌遗传学与生物学》30:17,2000年。(3)S. Danielson等人,《种子科学与技术》32:91,2004年(4)J. Ochoa等人,《植物病理学》48:425,1999年。