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加纳首次报道由大豆锈菌引起的大豆锈病

First Report of Soybean Rust Caused by Phakopsora pachyrhizi in Ghana.

作者信息

Bandyopadhyay R, Ojiambo P S, Twizeyimana M, Asafo-Adjei B, Frederick R D, Pedley K F, Stone C L, Hartman G L

机构信息

International Institute of Tropical Agriculture (IITA), PMB 5320, Ibadan, Nigeria.

Crop Research Institute, Kumasi, Ghana.

出版信息

Plant Dis. 2007 Aug;91(8):1057. doi: 10.1094/PDIS-91-8-1057B.

Abstract

Nigeria is the only country in West Africa where soybean rust, caused by Phakopsora pachyrhizi, has been officially reported (1). During a disease survey in Ghana during October 2006, soybean (Glycine max) leaves with rust symptoms (tan, angular lesions with erumpent sori exuding urediniospores) were observed in 11 fields in the following districts: Kassena Nankana in the Upper East Region; East Gonja, Central Gonja, and Tolon-Kumbungu in the Northern Region; and Ejisu-Juabeng in the Ashanti Region. Disease incidence in these fields ranged from 50 to 100% and disease severity ranged between 3 and 40% of the leaf area on infected plants. Urediniospores were hyaline, minutely echinulate, and 23 to 31 × 14 to 18 μm. Within a week of collection, leaf samples were sent to the USDA-ARS Foreign Disease-Weed Science Research Unit for verification of pathogen identity. DNA was extracted from leaf pieces containing sori with the Qiagen DNeasy Plant Mini kit (Valencia, CA), and all 11 field samples amplified in a real-time fluorescent PCR with the P. pachyrhizi-specific primers Ppm1 and Ppa2 (2). Sequence alignment of the internal transcribed spacer (ITS) region 2 further confirmed the identification as P. pachyrhizi (2). Infected leaves from three fields were separately washed in sterile water to collect urediniospores that were used to separately inoculate three detached leaves (for each isolate) of susceptible cultivar TGx 1485-1D (3). The abaxial surface of detached leaves was sprayed with 400 μl of spore suspension (1 × 10 spores per ml). A single leaf piece was placed in a 9-cm-diameter petri dish with adaxial side appressed on 1% technical agar amended with 10 μg/ml of kinetin. Lactic acid (1.5 ml/liter) and benomyl (12.5 mg/liter) were added to the agar medium to inhibit growth of saprophytic fungi and bacteria. Petri dishes were incubated at 20°C with a 12-h light/12-h dark cycle. Lesions on inoculated leaves developed 5 to 6 days after inoculation (DAI), and pustules (105 to 120 μm) formed 7 to 8 DAI and erupted 3 days later exuding columns of urediniospores similar in size to the initially collected isolates. Inoculating another set of detached leaves with a spore suspension (1 × 10 spores per ml) from the first set of detached leaves resulted in typical rust symptoms. The PCR assay, alignment of ITS region 2, morphological characters of the isolates, and pathogenicity tests demonstrate that P. pachyrhizi occurs in Ghana. To our knowledge, this is the first report of P. pachyrhizi in Ghana. References: (1) O. A. Akinsanmi et al. Plant Dis. 85:97, 2001. (2) R. D. Frederick et al. Phytopathology 92:217, 2002. (3) M. Twizeyimana et al. Online publication. http://www.plantmanagementnetwork.org/ infocenter/topic/soybeanrust/2006/posters/41.asp. Plant Management Network, 2006.

摘要

尼日利亚是西非唯一官方报道过由大豆锈菌(Phakopsora pachyrhizi)引起大豆锈病的国家(1)。在2006年10月于加纳进行的一次病害调查中,在以下地区的11块田地中观察到有锈病症状(棕褐色、具角状病斑且有突出的夏孢子堆并散发出夏孢子)的大豆(Glycine max)叶片:上东部地区的卡塞纳·南卡纳;北部地区的东戈贾、中戈贾和托隆 - 昆本古;以及阿散蒂地区的埃吉苏 - 朱阿本。这些田地中的发病率为50%至100%,病情严重程度在受感染植株叶片面积的3%至40%之间。夏孢子无色透明,具微小刺状突起,大小为23至31×14至18μm。采集后一周内,叶片样本被送往美国农业部农业研究局外来病害 - 杂草科学研究室以验证病原菌身份。使用Qiagen DNeasy植物微量提取试剂盒(加利福尼亚州瓦伦西亚)从含有夏孢子堆的叶片碎片中提取DNA,并且所有11个田间样本都用大豆锈菌特异性引物Ppm1和Ppa2进行实时荧光PCR扩增(2)。内部转录间隔区(ITS)2区域的序列比对进一步确认其为大豆锈菌(2)。从三块田地中采集的感染叶片分别在无菌水中冲洗以收集夏孢子,这些夏孢子被用于分别接种易感品种TGx 1485 - 1D的三片离体叶片(每个分离株)(3)。在离体叶片的背面喷洒400μl孢子悬浮液(每毫升含1×10个孢子)。将一片叶片碎片放入直径9厘米的培养皿中,正面贴在添加了10μg/ml激动素的1%工业琼脂上。向琼脂培养基中添加乳酸(1.5毫升/升)和苯菌灵(12.5毫克/升)以抑制腐生真菌和细菌的生长。培养皿在20°C下、12小时光照/12小时黑暗周期条件下培养。接种叶片上的病斑在接种后5至6天出现,脓疱(105至120μm)在接种后7至8天形成,并在3天后破裂,散发出与最初采集的分离株大小相似的夏孢子柱。用来自第一组离体叶片的孢子悬浮液(每毫升含1×10个孢子)接种另一组离体叶片会产生典型的锈病症状。PCR检测、ITS区域2的比对、分离株的形态特征以及致病性测试表明大豆锈菌在加纳存在。据我们所知,这是大豆锈菌在加纳的首次报道。参考文献:(1)O. A. Akinsanmi等人,《植物病害》85:97,2001年。(2)R. D. Frederick等人,《植物病理学》92:217,2002年。(3)M. Twizeyimana等人,在线出版物。http://www.plantmanagementnetwork.org/ infocenter/topic/soybeanrust/2006/posters/41.asp。植物管理网络,2006年。

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