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疫霉属病菌引起的七瓣莲叶片感染的首次报告

First Report of Foliar Infection of Starflower by Phytophthora ramorum.

作者信息

Hüberli D, Van Sant-Glass W, Tse J G, Garbelotto M

机构信息

Department of ESPM-ES, 151 Hilgard Hall, University of California, Berkeley 94720.

出版信息

Plant Dis. 2003 May;87(5):599. doi: 10.1094/PDIS.2003.87.5.599B.

Abstract

In March 2002, Phytophthora ramorum S. Werres & A.W.A.M. de Cock was isolated from pacific or western starflower (Trientalis latifolia Hook.), an herbaceous perennial of the Primulaceae family, at Castro Canyon in Big Sur, Monterey County, California. Affected leaves had numerous necrotic lesions >5 mm in diameter surrounded by a yellow halo, and the lesions coalesced with time. Isolates were identified as P. ramorum by the large chlamydospores, caduceus, semipapillate sporangia, and sequences of the internal transcribed spacer (ITS) region of the rDNA (1,2). The same symptoms were observed on starflower in a second location at the Soquel Demonstration Forest, Santa Cruz County. Although P. ramorum was not isolated from symptomatic leaves on the plants in Santa Cruz County, the ITS region of the pathogen was amplified and sequenced using P. ramorum-specific primers. Both sites were mixed forests of coast redwood (Sequoia sempervirens), bay laurel (Umbellularia californica), and tanoak (Lithocarpus densiflorus), which are confirmed hosts of P. ramorum. To test for pathogenicity to starflower, asymptomatic plants were carefully excavated from the two forest locations, replanted in 15-cm paper cups in the original forest soil, and the foliage was inoculated with zoospores of P. ramorum isolate Pr-52, an isolate used in previous inoculations. The zoospores were produced by placing agar disks (1 cm in diameter) from the margin of 8- to 14-day-old colonies growing on V8 juice agar into 20 to 30 ml of sterile deionized water in petri dishes. After 2 days incubation at 20°C in the dark, zoospore release was induced by placing dishes at 4°C for 20 min and then to room temperature for 45 to 60 min. Three hundred μl of the zoospore suspension (approximately 2 × 10 zoospores/ml) was poured into 500-μl modified microcentrifuge tubes in which tips of leaves of starflower were submerged. Control leaves were dipped in sterile deionized water. Plants were placed in a humid-chamber consisting of moist paper towels placed on the tray and covered with a clear-plastic lid that was sprayed with sterile water. The chambers were maintained at 20 to 24°C in the laboratory. Two or three leaves were inoculated, and one leaf was left as the control on each of seven or eight plants in two separate trials. In both trials, water-soaked lesions were observed on the leaves 12 h after inoculation with P. ramorum. At 8 or 11 days after inoculation, necrotic lesions were present on all inoculated leaves starting from the leaf tips. Lesions averaged 29 mm (range 13 to 39 mm) and 45 mm (range 31 to 56 mm) in length in the respective trials. Some lesions covered entire leaves. P. ramorum was reisolated on Phytophthora-selective agar medium (1) from the lesions in both trials. Control leaves had no lesions, and P. ramorum was not reisolated. Infection of starflower and other understory species appears to occur under infested tree hosts such as bay laurel, which is known as a source of inoculum for P. ramorum. To our knowledge, this is the first report of an herbaceous host for P. ramorum and the first report of the disease on the Primulaceae. Previously, only woody hosts were known. Starflower is unlikely to play a major role in the natural spread of the disease, but the pathogen may be spread via movement of plants through the horticultural industry. Furthermore, Trientalis spp. in Europe where P. ramorum is present may also be potential hosts. References: (1) D. M. Rizzo et al. Plant Dis. 86:205, 2002. (2) S. Werres et al. Mycol. Res. 105:1155, 2001.

摘要

2002年3月,在加利福尼亚州蒙特雷县大苏尔的卡斯特罗峡谷,从报春花科多年生草本植物太平洋七瓣莲(Trientalis latifolia Hook.)中分离出了樟疫霉(Phytophthora ramorum S. Werres & A.W.A.M. de Cock)。受感染的叶片上有许多直径大于5毫米的坏死病斑,病斑周围有黄色晕圈,且病斑会随着时间融合。通过大型厚垣孢子、双轮体、半乳头状孢子囊以及核糖体DNA(rDNA)的内转录间隔区(ITS)序列,将分离菌株鉴定为樟疫霉(1,2)。在圣克鲁斯县索奎尔示范森林的另一个地点,七瓣莲上也观察到了相同症状。尽管在圣克鲁斯县的植株上,未从有症状的叶片中分离出樟疫霉,但使用樟疫霉特异性引物对病原菌的ITS区域进行了扩增和测序。这两个地点均为海岸红杉(Sequoia sempervirens)、月桂树(Umbellularia californica)和鞣皮栎(Lithocarpus densiflorus)的混交林,这些都是已确认的樟疫霉寄主。为测试对七瓣莲的致病性,从这两个森林地点小心挖出无症状植株,重新种植在装有原始森林土壤的15厘米纸杯里,并用樟疫霉分离株Pr - 52的游动孢子接种其叶片,该分离株曾用于之前的接种试验。游动孢子通过将直径1厘米的琼脂圆盘从在V8汁琼脂上生长8至14天的菌落边缘放置到培养皿中的20至30毫升无菌去离子水中产生。在黑暗中20°C培养2天后,通过将培养皿置于4°C 20分钟,然后再置于室温45至60分钟来诱导游动孢子释放。将300微升游动孢子悬浮液(约2×10个游动孢子/毫升)倒入500微升改良微量离心管中,七瓣莲叶片尖端浸入其中。对照叶片浸入无菌去离子水中。植株放置在一个潮湿箱中,潮湿箱由放置在托盘上并用喷有无菌水的透明塑料盖覆盖湿纸巾组成。这些箱子在实验室中保持在20至24°C。在两个独立试验中,对七八株植物中的两三片叶子进行接种,每株留一片叶子作为对照。在两个试验中,接种樟疫霉12小时后,叶片上均观察到水渍状病斑。接种后8或11天,从叶尖开始,所有接种叶片上都出现了坏死病斑。在各自试验中,病斑长度平均为29毫米(范围13至39毫米)和45毫米(范围31至56毫米)。一些病斑覆盖了整片叶子。在两个试验中,均从病斑中在疫霉选择性琼脂培养基(1)上重新分离出樟疫霉。对照叶片无病斑,也未重新分离出樟疫霉。七瓣莲和其他林下物种的感染似乎发生在受侵染的树木寄主如月桂树之下,月桂树是已知的樟疫霉接种源。据我们所知

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