Caesar A J, Lartey R T
USDA, ARS, Northern Plains Agricultural Research Lab, 1500 North Central Avenue, Sidney, MT 59270.
Plant Dis. 2009 Aug;93(8):846. doi: 10.1094/PDIS-93-8-0846A.
The exotic, rangeland weed Lepidium draba L., a brassicaceous perennial, is widely distributed in the United States. For example, Oregon contains 100,000 ha of land infested with L. draba (2). Because it is capable of aggressive spread and has the potential to reduce the value of wheat-growing land (4), it is the target of biological control research. The application of multiple pathogens has been advocated for control of other brassicaceous weeds, including the simultaneous application of biotrophic and necrotrophic pathogens (3). In pursuit of this approach, in 2007, we discovered the occurrence of leaf spots on approximately 90% of L. draba plants near Shepherd, MT, which were distinct from leaf lesions caused by Cercospora bizzozeriana (1). The lesions were initially tiny, black spots enlarging over time to become circular to irregular and cream-colored around the initial black spots and sometimes with dark brown borders or chlorotic halos. Conidia from the lesions were light brown, elongate and obclavate, produced singly from short conidia, with 8 to 12 transverse septa, and 2 to 6 longitudinal septa. The spore body measured 25 to 35 × 200 to 250 μm with a beak cell 42 to 100 μm long. On the basis of conidial and cultural characteristics, the fungus was identified as Alternaria brassicae (Berk.) Sacc. Leaf tissues bordering lesions were plated on acidified potato dextrose agar. Colonies on V8 and alfalfa seed agar were black with concentric rings, eventually appearing uniformly black after 10 to 14 days. The internal transcribed spacer region of rDNA was amplified using primers ITS1 and ITS4 and sequenced. BLAST analysis of the 575-bp fragment showed a 100% homology with a sequence of A. brassicae Strain B from mustard (GenBank Accession No. DQ156344). The nucleotide sequence has been assigned GenBank Accession No. FJ869872. For pathogenicity tests, aqueous spore suspensions approximately 10/ml were prepared from cultures grown at 20 to 25°C for 10 to 14 days on V8 agar and sprayed on leaves of three L. draba plants. Inoculated plants were enclosed in plastic bags and incubated at 20 to 22°C for 72 to 80 h. In addition, three plants of the following reported hosts of A. brassicae were inoculated: broccoli, canola, Chinese cabbage, collards, broccoli raab, kale, mustard greens, radish, rape kale, and turnip. Within 10 days, leaf spots similar to those described above developed on plants of radish, canola, Chinese cabbage, and turnip and A. brassicae was reisolated and identified. Control plants sprayed with distilled water remained symptomless. These inoculations were repeated and results were the same. To our knowledge, this is the first report of a leaf spot disease caused by A. brassicae on L. draba in North America. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI No. 878750A). References: (1) A. J. Caesar et al. Plant Dis. 93:108, 2009. (2) G. L. Kiemnec and M. L. McInnis. Weed Technol. 16:231, 2002. (3) A. Maxwell and J. K. Scott. Adv. Bot. Res. 43:143, 2005. (4) G. A. Mulligan and J. N. Findlay. Can. J. Plant Sci. 54:149, 1974.
外来的牧场杂草——十字花科多年生植物白芥(Lepidium draba L.),在美国分布广泛。例如,俄勒冈州有10万公顷土地受到白芥的侵害(2)。由于它能够迅速蔓延,并有可能降低小麦种植土地的价值(4),因此成为生物防治研究的目标。有人主张应用多种病原体来控制其他十字花科杂草,包括同时应用活体营养型和死体营养型病原体(3)。为了探索这种方法,2007年我们在蒙大拿州谢泼德附近发现约90%的白芥植株出现叶斑,这些叶斑与油菜尾孢菌(Cercospora bizzozeriana)引起的叶斑不同(1)。病斑最初是微小的黑点,随着时间的推移会扩大,变成圆形至不规则形状,在最初的黑点周围呈米色,有时有深褐色边缘或褪绿晕圈。病斑上的分生孢子浅褐色,细长且倒棒状,单生于短分生孢子梗上,有8至12个横隔膜和2至6个纵隔膜。孢子体长25至35×200至250μm,喙细胞长42至100μm。根据分生孢子和培养特征,该真菌被鉴定为芸苔链格孢(Alternaria brassicae (Berk.) Sacc.)。将病斑边缘的叶片组织接种在酸化的马铃薯葡萄糖琼脂上。在V8琼脂和苜蓿种子琼脂上的菌落为黑色,有同心环,10至14天后最终变为均匀黑色。使用引物ITS1和ITS4扩增rDNA的内部转录间隔区并进行测序。对575bp片段的BLAST分析显示,与来自芥菜的芸苔链格孢菌株B的序列有100%的同源性(GenBank登录号:DQ156344)。该核苷酸序列已被赋予GenBank登录号:FJ869872。为进行致病性测试,从在V8琼脂上于20至25°C培养10至14天的培养物中制备约10个/ml的水悬浮孢子液,并喷洒在三株白芥植株的叶片上。接种后的植株用塑料袋套住,在20至22°C下培养72至80小时。此外,还对接种了以下芸苔链格孢已知寄主的三株植物进行了接种:西兰花、油菜、大白菜、羽衣甘蓝、抱子甘蓝、羽衣甘蓝、芥菜叶、萝卜、羽衣甘蓝苔和芜菁。在10天内,萝卜、油菜、大白菜和芜菁植株上出现了与上述描述相似的叶斑,并且再次分离并鉴定出芸苔链格孢。喷洒蒸馏水的对照植株无症状。这些接种实验重复进行,结果相同。据我们所知,这是北美地区关于芸苔链格孢引起白芥叶斑病的首次报道。一份凭证标本已保存在美国国家真菌收藏中心(BPI编号:878750A)。参考文献:(1)A. J. Caesar等人,《植物病害》93:108,2009年。(2)G. L. Kiemnec和M. L. McInnis,《杂草技术》16:231,2002年。(3)A. Maxwell和J. K. Scott,《植物学研究进展》43:143,2005年。(4)G. A. Mulligan和J. N. Findlay,《加拿大植物科学杂志》54:149,1974年。