Huang H C, Erickson R S, Van Hezewijk B, De Clerck-Floate R
Agriculture and Agri-Food Canada Research Centre, Lethbridge.
Plant Dis. 2005 Sep;89(9):1013. doi: 10.1094/PD-89-1013C.
Houndstongue (Cynoglossum officinale L.) is a rangeland weed introduced into Canada from Eurasia, and it can be highly toxic to livestock feeding in pastures (3). During 2004, houndstongue plants near Creston, BC, Canada developed water-soaked lesions with white mycelia and black sclerotia on leaves and crowns. Isolations from diseased leaf tissues and sclerotia on potato dextrose agar (PDA) at 20°C for 5 to 7 days produced fungal colonies with formation of black sclerotia 5 to 10 mm in diameter. A single hyphal tip isolate from houndstongue. Ss-HT-C. was compared with a sunflower isolate of S. sclerotiorum, sun-87 (1), for morphology and pathogenicity. For apothecial production, Ss-HT-C and sun-87 were grown on PDA in petri dishes at 10°C for 10 weeks, and sclerotia produced were harvested, placed on moist vermiculite in petri dishes, and incubated at 20°C under light for 3 weeks. Mature apothecia were excised, stained with acid fuchsin, mounted on slides, and examined for asci and ascospores with a microscope. There were no morphological differences between Ss-HT-C and sun-87, each producing an ascus with eight binucleate, elliptical ascospores, measuring 4 × 10 μm (width × length), supporting the identity of Ss-HT-C as S. sclerotiorum (2,4). For pathogenicity tests of Ss-HT-C and sun-87, mycelial plugs (8 mm in diameter) were removed from the margin of colonies grown on PDA for 5 days at 20°C, and placed on leaves of C. officinale plants that were grown in a greenhouse (20 ± 4°C) to the rosette stage. Inoculated plants were covered with clear plastic bags, kept in the same greenhouse for 3 days, and the diameters of the leaf lesions developed at inoculation sites were measured. The experiment was run twice with 30 plants per isolate and five leaves per plant. Uninoculated plants covered with plastic bags were used as controls. Experiments used a completely randomized design. Results of leaf inoculations showed that Ss-HT-C and sun-87 were pathogenic to hound-stongue. There was no statistical difference between isolates or trials. The frequency of leaves with lesions was 90% for Ss-HT-C and 93% for sun-87. The mean leaf lesion diameters were 32 and 35 mm for Ss-HT-C and sun-87, respectively. Leaves of control plants remained healthy. S. sclerotiorum was reisolated from leaves with lesions, but not from controls. After 14 to 21 days, new sclerotia, 5 to 10 mm in diameter, were formed on leaves of inoculated plants. The plants eventually died. This study confirms that S. sclerotiorum is the causal agent for the disease of hound-stongue in Canada, and to our knowledge, this is the first world record of infection of this weed by S. sclerotiorum. References: (1) H. C. Huang and G. C. Kozub. Plant Prot. Bull. 31:333, 1989. (2) L. Kohn, Phytopathology 69:881, 1979. (3) J. A. Pfister et al. J. Range Manag. 45:254, 1992. (4) J. A. L. Wong and H. J. Willetts, J. Gen. Microbiol. 112:29, 1979.
犬舌草(Cynoglossum officinale L.)是一种从欧亚大陆传入加拿大的牧场杂草,对在牧场觅食的牲畜具有高毒性(3)。2004年期间,在加拿大不列颠哥伦比亚省克雷斯顿附近的犬舌草植株在叶片和叶冠上出现了有水渍状病斑,带有白色菌丝体和黑色菌核。在20°C下于马铃薯葡萄糖琼脂(PDA)上从患病叶片组织和菌核进行分离培养5至7天,产生了带有直径5至10毫米黑色菌核形成的真菌菌落。从犬舌草分离得到的一个单菌丝尖端分离物Ss-HT-C,与核盘菌的一个向日葵分离物sun-87(1)进行了形态学和致病性比较。为了产生子囊盘,将Ss-HT-C和sun-87在培养皿中的PDA上于10°C培养10周,收获产生的菌核,放置在培养皿中潮湿的蛭石上,并在20°C光照下培养3周。切下成熟的子囊盘,用酸性品红染色,装片在显微镜下检查子囊和子囊孢子。Ss-HT-C和sun-87之间没有形态学差异,每个都产生一个带有八个双核、椭圆形子囊孢子的子囊,尺寸为4×10微米(宽×长),支持将Ss-HT-C鉴定为核盘菌(2,4)。对于Ss-HT-C和sun-87的致病性测试,从在20°C下于PDA上培养5天的菌落边缘切下直径8毫米的菌丝块,放置在温室(20±4°C)中生长至莲座期的犬舌草植株的叶片上。接种的植株用透明塑料袋覆盖,在同一温室中放置3天,测量接种部位形成的叶片病斑直径。该实验重复进行两次,每个分离物用30株植物,每株植物5片叶子。用覆盖塑料袋的未接种植株作为对照。实验采用完全随机设计。叶片接种结果表明,Ss-HT-C和sun-87对犬舌草具有致病性。分离物或试验之间没有统计学差异。Ss-HT-C处理的叶片病斑出现频率为90%,sun-87为93%。Ss-HT-C和sun-87处理的叶片平均病斑直径分别为32毫米和35毫米。对照植株的叶片保持健康。从有病斑的叶片上重新分离到了核盘菌,但对照叶片上未分离到。14至21天后,在接种植株的叶片上形成了直径5至10毫米的新菌核。植株最终死亡。本研究证实核盘菌是加拿大犬舌草病害的病原菌,据我们所知,这是该杂草被核盘菌感染的首个世界纪录。参考文献:(1)H.C. Huang和G.C. Kozub。植物保护通报31:333,1989。(2)L. Kohn,植物病理学69:881,1979。(3)J.A. Pfister等人。牧场管理杂志45:254,1992。(4)J.A.L. Wong和H.J. Willetts,普通微生物学杂志112:29,1979。