Petróczy M, Csejk Gy, Palkovics L
Department of Plant Pathology, Corvinus University of Budapest, Ménesi Road 44, H-1118 Budapest, Hungary. The project was funded by TÁMOP- 4.2.1./B-09/1-KMR-2010-0005 and TÁMOP- 4.2.2./B-10/1-2010-0023 grants.
Plant Dis. 2012 Jan;96(1):148. doi: 10.1094/PDIS-07-11-0623.
In May 2011, young impatiens plants (Impatiens walleriana Hook.f.) showing downy mildew symptoms were collected from a greenhouse in Kecskemét, Hungary. The disease was later reported from different parts of the country from a number of different cultivars. The upper surface of affected leaves turned yellow and white fungal-like growth was observed on the underside. Diseased plants wilted and rapidly collapsed, resulting in losses of nearly 100%. Appearance of the disease caused a loss of approximately 2,000 euros for the growers in Kecskemét. In Hungary, losses for growers and consumers could have reached half a million euros. Downy mildew of impatiens can be caused by two pathogens, Plasmopara obducens or Bremiella sphaerosperma (1). P. obducens differs from B. sphaerosperma by monopodial sporangiophores with straight branches and the lack of apical thickening on the branchlets. Sporangia of B. sphaerosperma are spherical, while those of P. obducens are subspherical. Sporangiophores of the fungus were colorless with straight trunks and monopodially branched four to seven times. Sporangiophores ended with three apical branchlets at right angles to the main axis, measuring 4.6 to 16.4 μm (average 9.6 μm). The ovoid and hyaline sporangia measured 11.04 (7.7 to 13.8) × 13.9 (9.9 to 17.4) μm. For molecular identification, total DNA was extracted from the pathogen scraped from the leaves using a cetyltrimethylammoniumbromide (CTAB) extraction method. The 5'-end of the large subunit of ribosomal RNA gene was amplified by PCR using NL1 and NL4 primers (3) and cloned and sequenced. The nucleotide sequence of IWPO-H1 Hungarian isolate (GenBank Accession No. HE577169) showed highest identity with Accession Nos. AY587558 and HQ246451 from the United Kingdom and Serbia (2), respectively, with 99.72% identity. On the basis of the symptoms and molecular and morphological characters, the pathogen was identified as P. obducens. Pathogenicity was confirmed by spraying young I. walleriana plants with a sporangial suspension (1 × 10 sporangiospores ml). Control plants were sprayed with sterile water. Plants were incubated at room temperature under glass vessels at approximately 90% relative humidity. Symptoms of downy mildew occurred 12 to 16 days after inoculation, while control plants remained healthy. Presence of P. obducens was reported from Austria, Australia, Bulgaria, China, the Czech Republic, Denmark, Germany, Finland, Korea, Lithuania, Norway, Pakistan, Poland, Romania, Russia, the United Kingdom, the United States, and recently from Serbia (2). To our knowledge, this is the first report of downy mildew of I. walleriana caused by P. obducens in Hungary. The appearance of P. obducens in Hungary seriously endangers the production of I. walleriana, which is an important and popular ornamental in gardens and city parks. References: (1) O. Constantinescu. Mycologia 83:473, 1991. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , September 1, 2011. (3) K. L. O'Donnell. The Fungal Holomorph: Mitotic, Meiotic and Pleomorphic Speciation in Fungal Systematics. CAB International, Wallingford, UK, 1993.
2011年5月,从匈牙利凯奇凯梅特的一个温室中采集到表现霜霉病症状的凤仙花嫩苗(凤仙花)。该病随后在该国不同地区的多个不同品种上被发现。患病叶片的上表面变黄,下表面可见类似真菌的白色生长物。患病植株枯萎并迅速倒伏,损失近100%。该病的出现给凯奇凯梅特的种植者造成了约2000欧元的损失。在匈牙利,种植者和消费者的损失可能达到50万欧元。凤仙花霜霉病可由两种病原体引起,即凤仙花盘梗霉或球孢盘梗霉(1)。凤仙花盘梗霉与球孢盘梗霉的区别在于,前者的孢囊梗单轴分枝,分枝直,小枝顶端无加厚;球孢盘梗霉的孢子囊呈球形,而凤仙花盘梗霉的孢子囊呈近球形。该真菌的孢囊梗无色,主干直,单轴分枝4至7次。孢囊梗末端有3个与主轴成直角的顶端小枝,长4.6至16.4μm(平均9.6μm)。卵形透明的孢子囊大小为11.04(7.7至13.8)×13.9(9.9至17.4)μm。为进行分子鉴定,使用十六烷基三甲基溴化铵(CTAB)提取法从叶片上刮取的病原体中提取总DNA。使用NL1和NL4引物(3)通过PCR扩增核糖体RNA基因大亚基的5′端,并进行克隆和测序。匈牙利分离株IWPO - H1的核苷酸序列(GenBank登录号HE577169)与来自英国和塞尔维亚的登录号AY587558和HQ246451的序列分别具有最高同源性,同源性为99.72%。根据症状以及分子和形态学特征,该病原体被鉴定为凤仙花盘梗霉。通过向凤仙花嫩苗喷洒孢子囊悬浮液(1×10个孢子囊/毫升)来确认致病性。对照植株喷洒无菌水。将植株在室温下置于玻璃容器中,相对湿度约为90%。接种后12至16天出现霜霉病症状,而对照植株保持健康。奥地利、澳大利亚、保加利亚、中国、捷克共和国、丹麦、德国、芬兰、韩国、立陶宛、挪威、巴基斯坦、波兰、罗马尼亚、俄罗斯、英国、美国以及最近的塞尔维亚均有凤仙花盘梗霉的报道(2)。据我们所知,这是匈牙利首次报道由凤仙花盘梗霉引起的凤仙花霜霉病。凤仙花盘梗霉在匈牙利的出现严重危及凤仙花的生产,凤仙花是花园和城市公园中一种重要且受欢迎的观赏植物。参考文献:(1)O. Constantinescu. Mycologia 83:473, 1991.(2)D. F. Farr和A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. 从http://nt.ars-grin.gov/fungaldatabases/获取,2011年9月1日。(3)K. L. O'Donnell. The Fungal Holomorph: Mitotic, Meiotic and Pleomorphic Speciation in Fungal Systematics. CAB International, Wallingford, UK, 1993.