Lanoiselet V, You M P, Li Y P, Wang C P, Shivas R G, Barbetti M J
Department of Agriculture and Food Western Australia, Baron-Hay Court, South Perth, W.A. 6151, Australia and School of Plant Biology, The University of Western Australia, Crawley, W.A. 6009 Australia.
School of Plant Biology, The University of Western Australia, Crawley, W.A. 6009, Australia.
Plant Dis. 2012 Sep;96(9):1382. doi: 10.1094/PDIS-04-12-0415-PDN.
Rice (Oryza sativa L.) has been grown in the Ord River Irrigation Area (ORIA) in northern Western Australia since 1960. In 2011, a sheath rot of rice was observed in the ORIA. Symptoms were variable, appearing as either (i) oblong pale to dark brown lesions up to 3 cm length, (ii) lesions with pale grey/brown centers and with dark brown margins, or (iii) diffuse dark or reddish brown streaks along the sheath. Lesions enlarged and coalesced, often covering the majority of the leaf sheath, disrupting panicle emergence. Isolations from small pieces of infested tissues from plants showing sheath rot symptoms were made onto water agar, subcultured onto potato dextrose agar, cultures maintained at 20°C, and a representative culture lodged both in the Western Australian Culture Collection maintained at the Department of Agriculture and Food Western Australia (as WAC 13481) and in the culture collection located at the DAFF Plant Pathology Herbarium (as BRIP 54763). Amplification of the internal transcribed spacer (ITS)1 and (ITS)2 regions flanking the 5.8S rRNA gene were carried out with universal primers ITS1 and ITS4 according to the published protocol (4). The DNA PCR products from a single isolate were sequenced and BLAST analyses used to compare sequences with those in GenBank. The sequence had 99% nucleotide identity with the corresponding sequence in GenBank for Sarocladium oryzae (Sawada) W. Gams & D. Hawksworth. Isolates showed morphological (e.g., conidiophore and conidia characteristics) (2) and molecular (1) similarities with S. oryzae as described in other reports. The relevant sequence information for a representative isolate was lodged in GenBank (GenBank Accession No. JQ965668). Spores of S. oryzae were produced on rice agar under "black light" at 22°C to induce sporulation over 4 weeks. Under conditions of 30/28°C (day/night), 14/12 h (light/dark), rice cv. Quest, grown for 11 weeks until plants reached the tillering stage, was inoculated by spraying a suspension 5 × 10 spores/ml of the same single isolate onto foliage until runoff occurred. Inoculated plants were placed under a dark plastic cover for 72 h to maximize humidity levels around leaves and subsequently maintained under >90% relative humidity conditions. Symptoms of sheath rot as described in (i) and (ii) above appeared by 14 days after inoculation, with lesions up to 23 cm long by 15 days post-inoculation. Severe disease prevented young panicles from emerging. Infection studies were successfully repeated and S. oryzae was reisolated from leaf lesions 1 week after lesion appearance. No disease was observed on water-inoculated control rice plants. There have been records of S. oryzae on rice in New South Wales in the early 1980s (3) and in 2006 to 2007 (Australian Plant Pest Database), but to our knowledge, this is the first report of this pathogen in Western Australia. References: (1) N. Ayyadurai et al. Cur. Microbiol. Mycologia 50:319, 2005. (2) B. L. K. Brady. No. 673 in: IMI Descriptions of Fungi and Bacteria, 1980. (3) D. Phillips et al. FAO Plant Prot. Bull. 40:4, 1992. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.
自1960年以来,水稻(Oryza sativa L.)一直在西澳大利亚州北部的奥德河灌溉区(ORIA)种植。2011年,在ORIA观察到水稻鞘腐病。症状多样,表现为:(i)长达3厘米的长方形淡褐色至深褐色病斑;(ii)病斑中心为淡灰/褐色、边缘为深褐色;或(iii)沿着叶鞘的弥漫性暗褐色或红褐色条纹。病斑扩大并融合,常常覆盖大部分叶鞘,阻碍穗的抽出。从表现出鞘腐病症状的植株上取小块受侵染组织,分离培养于水琼脂上,继代培养于马铃薯葡萄糖琼脂上,培养物在20°C下保存,一份代表性培养物保存在西澳大利亚州农业和食品部维护的西澳大利亚培养物保藏中心(编号为WAC 13481)以及保存在澳大利亚农业、渔业与林业部植物病理学标本馆的培养物保藏中心(编号为BRIP 54763)。根据已发表的方案(4),使用通用引物ITS1和ITS4对5.8S rRNA基因两侧的内转录间隔区(ITS)1和(ITS)2区域进行扩增。对单个分离株的DNA PCR产物进行测序,并使用BLAST分析将序列与GenBank中的序列进行比较。该序列与GenBank中稻帚枝霉(Sarocladium oryzae (Sawada) W. Gams & D. Hawksworth)的相应序列具有99%的核苷酸同一性。如其他报告所述分离株在形态学(如分生孢子梗和分生孢子特征)(2)和分子水平(1)上与稻帚枝霉相似。一份代表性分离株的相关序列信息保存在GenBank(GenBank登录号JQ965668)。在22°C的“黑光”下,在水稻琼脂上培养稻帚枝霉的孢子4周以诱导产孢。在30/28°C(日/夜)、14/12小时(光照/黑暗)条件下,种植水稻品种Quest,生长11周直至植株达到分蘖期,通过向叶片上喷洒浓度为5×10个孢子/ml的同一单个分离株的悬浮液直至径流发生来进行接种。对接种的植株用黑色塑料覆盖72小时以使叶片周围湿度最大化,随后保持在相对湿度>90%的条件下。接种后14天出现上述(i)和(ii)中描述的鞘腐病症状,接种后15天病斑长达23厘米。严重病害阻碍幼穗抽出。侵染研究成功重复,并且在病斑出现1周后从叶片病斑中重新分离到稻帚枝霉。在用水接种的对照水稻植株上未观察到病害。在20世纪80年代早期(3)以及2006至2007年(澳大利亚植物害虫数据库)新南威尔士州有水稻上稻帚枝霉的记录,但据我们所知,这是该病原菌在西澳大利亚州的首次报道。参考文献:(1) N. Ayyadurai等人,Cur. Microbiol. Mycologia 杂志50:319,2005年。(2) B. L. K. Brady,载于《IMI真菌和细菌描述》第673号,1980年。(3) D. Phillips等人,FAO植物保护通报40:4,1992年。(4) T. J. White等人,载于《PCR方案:方法与应用指南》第315页。学术出版社,加利福尼亚州圣地亚哥,1990年。