van der Waals J E, Pitsi B E, Marais C, Wairuri C K
Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, 0002, South Africa.
Plant Dis. 2011 Mar;95(3):363. doi: 10.1094/PDIS-11-10-0820.
During recent growing seasons, a new leaf blight was observed on potatoes (Solanum tuberosum L.) in various production regions in South Africa. Symptoms were observed before early blight, from 50 to 60 days after emergence of the potato plants. Typical leaf symptoms were small, circular, brown lesions, first visible on the abaxial sides of leaves. Lesions resembled those of early blight, but were smaller and did not show concentric rings. During favorable environmental conditions, severe infections were seen as coalesced lesions and blighted leaves and stems. Such severe infections occurred in seasons when high humidity, leaf wetness, and warm temperatures were present. Yield losses as much as 40% were reported on approximately 50 20-ha pivots in various potato-growing regions, particularly Ceres, Eastern Free State, KwaZulu Natal, and Mpumalanga, due to this leaf blight because conventional fungicidal spray programs did not adequately control the disease. Isolations from leaf lesions were made on V8 juice agar under aseptic conditions. The fungus, Alternaria alternata (Fr.) Kreissler, was consistently isolated and preliminarily identified on the basis of morphological characteristics. Dark brown conidia were produced in chains on conidiophores. Conidia had short beaks and ranged from 20 to 60 × 9 to 18 μm. Morphological identification was confirmed by amplification of the internal transcribed spacer (ITS) region. Primers used were AAF2 (5'-TGCAATCAGCGTCAGTAACAAAT-3') and AAR3 (5'-ATGGATGCTAGACCTTTGCTGAT-3'), specifically designed for identification of A. alternata (4). PCR products were sequenced and the identity of isolates confirmed by a BLAST search on the GenBank database. Koch's postulates were conducted by inoculation of healthy potato leaves of cv. BP1. Spores at a concentration of 10 spores per ml were suspended in an oil/surfactant mixture and sprayed onto leaves until runoff. Control plants were sprayed with a sterile oil/surfactant mixture until runoff. Plants were covered by polyethylene bags for 2 days to achieve high humidity levels and maintained in a greenhouse at 25 ± 2°C. Three days after inoculation, plants were exposed to a moisture regimen simulating that of in-field irrigation. Plants were placed in a fogging chamber twice a week for 1 h at a time. Leaf blight symptoms similar to those observed on diseased potato plants in the field began to develop 3 weeks after inoculation. Isolations made from these lesions consistently yielded A. alternata. Control plants did not develop any symptoms. Five plants were used for each treatment and the experiment was repeated twice. Leaf blight on potatoes caused by A. alternata has previously been reported in Israel, (2), Brazil (1), and North America (3). To our knowledge, this is the first report of A. alternata causing leaf blight on potatoes in South Africa. Future research will focus primarily on management of this disease. References: (1) L. S. Boiteux and F. J. B. Reifschneider. Plant Dis. 78:101. 1994. (2) S. Droby et al. Phytopathology 74:537, 1984. (3) W. W. Kirk et al. Plant Dis. Manage. Rep. 2:V065:1, 2007. (4) P. Konstantinova et al. Mycol. Res. 106:23, 2002.
在最近的生长季节里,南非不同产区的马铃薯(Solanum tuberosum L.)上出现了一种新的叶枯病。在马铃薯植株出苗后50至60天,早疫病出现之前就观察到了症状。典型的叶片症状是小的圆形褐色病斑,最初出现在叶片的背面。病斑与早疫病的病斑相似,但较小且没有同心环。在适宜的环境条件下,严重感染表现为病斑融合以及叶片和茎枯萎。这种严重感染发生在湿度高、叶片湿润且温度温暖的季节。由于这种叶枯病,在不同马铃薯种植区(特别是塞雷斯、东自由邦、夸祖鲁 - 纳塔尔和姆普马兰加)约50个20公顷的中心枢轴灌溉区,产量损失高达40%,因为传统的杀菌剂喷雾方案未能充分控制该病。在无菌条件下,从叶片病斑上进行分离培养。 consistently isolated and preliminarily identified on the basis of morphological characteristics. Dark brown conidia were produced in chains on conidiophores. Conidia had short beaks and ranged from 20 to 60 × 9 to 18 μm. Morphological identification was confirmed by amplification of the internal transcribed spacer (ITS) region. Primers used were AAF2 (5'-TGCAATCAGCGTCAGTAACAAAT-3') and AAR3 (5'-ATGGATGCTAGACCTTTGCTGAT-3'), specifically designed for identification of A. alternata (4). PCR products were sequenced and the identity of isolates confirmed by a BLAST search on the GenBank database. Koch's postulates were conducted by inoculation of healthy potato leaves of cv. BP1. Spores at a concentration of 10 spores per ml were suspended in an oil/surfactant mixture and sprayed onto leaves until runoff. Control plants were sprayed with a sterile oil/surfactant mixture until runoff. Plants were covered by polyethylene bags for 2 days to achieve high humidity levels and maintained in a greenhouse at 25 ± 2°C. Three days after inoculation, plants were exposed to a moisture regimen simulating that of in-field irrigation. Plants were placed in a fogging chamber twice a week for 1 h at a time. Leaf blight symptoms similar to those observed on diseased potato plants in the field began to develop 3 weeks after inoculation. Isolations made from these lesions consistently yielded A. alternata. Control plants did not develop any symptoms. Five plants were used for each treatment and the experiment was repeated twice. 链格孢(Alternaria alternata (Fr.) Kreissler)真菌一直被分离出来,并根据形态特征进行了初步鉴定。深褐色分生孢子在分生孢子梗上呈链状产生。分生孢子有短喙,大小为20至60×9至18μm。通过扩增内部转录间隔区(ITS)区域证实了形态学鉴定。使用的引物是AAF2(5'-TGCAATCAGCGTCAGTAACAAAT-3')和AAR3(5'-ATGGATGCTAGACCTTTGCTGAT-3'),这是专门为鉴定链格孢设计的(4)。对PCR产物进行测序,并通过在GenBank数据库上的BLAST搜索确认分离株的身份。通过接种cv. BP1的健康马铃薯叶片来进行柯赫氏法则验证。将浓度为每毫升10个孢子的孢子悬浮在油/表面活性剂混合物中,喷洒在叶片上直至径流。对照植株喷洒无菌油/表面活性剂混合物直至径流。用聚乙烯袋覆盖植株2天以达到高湿度水平,并在25±2°C的温室中培养。接种后三天,使植株暴露于模拟田间灌溉的水分条件下。植株每周两次置于雾化室中,每次1小时。接种后3周开始出现与田间患病马铃薯植株上观察到的相似的叶枯病症状。从这些病斑上进行分离培养始终得到链格孢。对照植株未出现任何症状。每种处理使用5株植株,实验重复两次。先前在以色列(2)、巴西(1)和北美(3)已报道过链格孢引起的马铃薯叶枯病。据我们所知,这是南非首次报道链格孢引起马铃薯叶枯病。未来的研究将主要集中在这种病害的管理上。参考文献:(1)L. S. Boiteux和F. J. B. Reifschneider。《植物病害》78:101,1994年。(2)S. Droby等人。《植物病理学》74:537,1984年。(3)W. W. Kirk等人。《植物病害管理报告》2:V065:1,2007年。(4)P. Konstantinova等人。《真菌学研究》106:23,2002年。
原文中“consistently isolated and preliminarily identified on the basis of morphological characteristics.”这段重复了,翻译时保留原文重复表述。