Agostini A, Johnson D A, Hulbert S, Demoz B, Fernando W G D, Paulitz T
Department of Plant Pathology, Washington State University, Pullman 99164-6430.
Dept. of Plant Science, University of Manitoba, Winnipeg, MB, Canada R3T 2N2.
Plant Dis. 2013 Jun;97(6):842. doi: 10.1094/PDIS-10-12-0956-PDN.
Canola (Brassica napus L.) is produced in the dryland agriculture areas of eastern Washington State and northern Idaho, often in rotation with cereal cropping systems. Canola is also used as a rotation crop in irrigated circles in the Columbia Basin of Washington and southern Idaho, where potato is the main cash crop. In 2011, 7,700 ha of canola were harvested in Idaho and 4,200 ha in Washington. One of the major diseases of canola around the world is blackleg, caused by Leptosphaeria maculans (aggressive) and L. biglobosa (non-aggressive). Both Washington and Idaho have been considered blackleg-free, and production of canola in Idaho is subject to government regulations. Canola seed originating from outside of Washington and Idaho should have a phytosanitary certificate. This disease is widespread in Canada and the U.S. Northern Plains, Midwest, and South, and is the major disease of canola in these areas. In August 2011, a sample from a canola field in Bonners Ferry, Idaho, was brought for diagnosis to Washington State University. The canola stems showed the typical gray to dark grey lesions with black pycnidia. The pycnidia and conidia were examined microscopically, and found to be similar to descriptions of Phoma lingam, the anamorph of L. maculans (2). Samples were sent to the University of Manitoba for confirmation with PCR. The pathogen was cultured out of stems on V8 juice agar amended with streptomycin and 22 single pynidiospore isolates were made from the cultures. DNA was extracted from the cultures using methods described in Fernando et al. (1) and a multiplex PCR was performed with species-specific primers for L. maculans and L. biglobosa. The reaction should produce a 330-bp amplicon for L. maculans and a 440-bp amplicon for L. biglobosa. Based on this, all 22 isolates were identified as L. maculans. The susceptible cultivar Westar was inoculated with the isolates, by wound inoculating 7-day-old cotyledons with a concentration of 10 spores/ml. Plants were kept in a moist chamber at 23°C. After 14 days, plants were rated for disease with a 0 to 9 scale, where 0 = no infection and 9 = tissue collapse and appearance of pycnidiospores. Isolates with rating ≥5 are considered virulent. All isolates produced a rating of 7 to 9, indicating a high level of virulence. The source of the seed used in the infested fields is not known at this time. This disease is seedborne, and may pose a threat to the two major vegetable and oilseed brassica seed production areas of Washington: the Skagit River valley of western Washington and the Columbia Basin area of central Washington. In addition, the susceptibility of Pacific Northwest varieties of canola and other brassica oilseeds is largely unknown. References: (1) W. G. D. Fernando et al. Plant Dis. 90:1337, 2006. (2) S. Roger Rimmer et al. Compendium of Brassica Diseases, APS Press, 2007.
油菜(甘蓝型油菜)产于华盛顿州东部和爱达荷州北部的旱地农业区,常与谷类作物种植系统轮作。在华盛顿州哥伦比亚盆地和爱达荷州南部的灌溉区,油菜也被用作轮作作物,那里马铃薯是主要经济作物。2011年,爱达荷州收获了7700公顷油菜,华盛顿州收获了4200公顷。油菜在全球的主要病害之一是黑胫病,由大茎点菌(致病型)和双生大茎点菌(非致病型)引起。华盛顿州和爱达荷州过去一直被认为无黑胫病,爱达荷州的油菜生产受政府监管。源自华盛顿州和爱达荷州以外地区的油菜种子应有植物检疫证书。这种病害在加拿大以及美国北部平原、中西部和南部广泛存在,是这些地区油菜的主要病害。2011年8月,爱达荷州邦纳斯费里一块油菜田的样本被送到华盛顿州立大学进行诊断。油菜茎上出现了典型的从灰色到深灰色的病斑,带有黑色分生孢子器。对分生孢子器和分生孢子进行显微镜检查,发现与大茎点菌的无性型——十字花科黑斑病菌的描述相符(2)。样本被送到曼尼托巴大学用聚合酶链式反应(PCR)进行确认。从茎上在添加链霉素的V8汁琼脂培养基上培养出病原菌,并从培养物中获得了22个单分生孢子分离株。按照费尔南多等人(1)所述方法从培养物中提取DNA,并用大茎点菌和双生大茎点菌的种特异性引物进行多重PCR。该反应应为大茎点菌产生一个330碱基对的扩增片段,为双生大茎点菌产生一个440碱基对的扩增片段。据此,所有22个分离株均被鉴定为大茎点菌。用这些分离株接种易感品种韦斯塔,方法是用浓度为每毫升10个孢子的菌液对7日龄子叶进行伤口接种。植株置于23℃的保湿箱中。14天后,按照0至9级对植株病害进行评级,0表示无感染,9表示组织坏死并出现分生孢子。评级≥5的分离株被视为有毒力。所有分离株的评级为7至9,表明毒力水平高。目前尚不清楚受侵染田块所用种子的来源。这种病害通过种子传播,可能对华盛顿州的两个主要蔬菜和油菜籽生产区构成威胁:华盛顿州西部的斯卡吉特河谷和华盛顿州中部的哥伦比亚盆地地区。此外,太平洋西北地区油菜品种和其他油菜籽的易感性在很大程度上尚不清楚。参考文献:(1)W.G.D.费尔南多等人,《植物病害》90:1337,2006年。(2)S.罗杰·里默等人,《十字花科病害简编》,美国植物病理学会出版社,2(此处疑似有误,应为2007)。