Cárdenas M E, Medina E, Tabima J, Vargas A, Lopera C, Bernal A, Restrepo S
Laboratorio de Micología y Fitopatología LAMFU, Universidad de los Andes, Carrera 1#18-10, J-205, Bogotá, Colombia.
Plant Dis. 2011 Jul;95(7):875. doi: 10.1094/PDIS-11-10-0853.
Solanum viarum Dunal (tropical soda apple) belongs to the section Acanthophora in the genus Solanum, which includes nearly 20 neotropical species of herbs and small shrubs (2). The species in this section are sometimes called the 'spiny Solanums' (2) and are adapted mainly to highly disturbed habitats and open secondary forests. The center of diversity is eastern Brazil (3). Since the early 1990s, S. viarum has been a problematic weed in Florida where it was listed as a noxious weed in 1993, followed in 1994 by its addition to the Federal Noxious Weed List of the USDA. On 17 April 2010, 12 plants of S. viarum located close to a S. betaceum crop (tree tomato) in the province of Caldas (Department of Antioquia, central northwestern Colombia) were found with symptoms similar to late blight caused by Phytophthora infestans on S. tuberosum (potato). Fifteen leaves from 12 plants with blackish, water-soaked lesions showing a white sporulation on the abaxial side were collected and processed within 3 days. The leaves were placed in a humid chamber and incubated in darkness at room temperature (18°C mean temperature) until sporulation was observed. Microscopic characteristics were consistent with Phytophthora spp. Only one axenic culture was obtained by successive subcultures in rye B agar plates. After an incubation period of 8 days, plates were washed with distilled water and ovoid, semipapillate caduceus sporangia ranging from 38 to 41 μm long (average 39; N = 86) and 23 to 29 μm wide (average 26; N = 86) were observed. To fulfill Koch's postulates and test the isolate for the ability to infect potato as well as Solanum spp. associated with potato crops in Colombia, triplicate pathogenicity tests were carried out on three detached leaves of S. viarum, S. tuberosum, and S. americanum (American nightshade). A 1 × 10 sporangia/ml suspension of the Phytophthora isolate, estimated using a haemocytometer, was obtained from 8-day-old cultures grown on rye B agar. The suspension was incubated at 4°C for 2 h to induce zoospore release. The leaves were then inoculated by spraying them until runoff. After an incubation period of 5 days at 18°C in a humidity chamber, mycelia, sporangia, and brownish lesions, similar to those described above, were observed in the leaves of all three hosts, indicating pathogenicity. DNA extraction was performed from the P. infestans isolate (4). Four nuclear loci, ITS, β-tubulin, Ras, and Avr3a, as well as one mitochondrial gene, cytochrome c oxidase 1 (Cox1), were amplified and sequenced. Sequences were compared with GenBank databases using Blastn. In all cases, the best hits corresponded to P. infestans (GenBank Accession No. HQ639930 for Avr3A, HQ639931 for β-tubulin, HQ639932 for Cox1, HQ639933 for iRas, HQ639934 for Ras, and JF419363 for ITS). Reports of P. infestans causing typical late blight symptoms on wild solanaceous plants are becoming more frequent and have been made from other countries such as Peru (1). To our knowledge, this is the first time that P. infestans has been observed and isolated from S. viarum in Colombia, introducing the possibility of this wild solanaceous weed as another late blight host. References: (1) G. Garry et al. Eur. J. Plant Pathol. 113:71, 2005. (2) R. Levin et al. Am. J. Bot. 92:603, 2005. (3) M. Nee. A Revision of Solanum Section Acanthophora. Ph.D. diss. University of Wisconsin, Madison, 1979. (4) A. M. Vargas et al. Phytopathology 99:82, 2009.
刺茄(Solanum viarum Dunal)属于茄属的刺萼茄组,该组包括近20种新热带地区的草本植物和小灌木(2)。该组中的物种有时被称为“多刺茄属植物”(2),主要适应于受高度干扰的栖息地和开阔的次生林。其多样性中心在巴西东部(3)。自20世纪90年代初以来,刺茄在佛罗里达州一直是一种有问题的杂草,1993年它被列为有害杂草,1994年又被列入美国农业部的联邦有害杂草名录。2010年4月17日,在卡尔达斯省(安蒂奥基亚省,哥伦比亚中西北部)靠近一株树番茄(S. betaceum)作物的地方发现了12株刺茄,其症状类似于马铃薯晚疫病,由致病疫霉(Phytophthora infestans)引起,发生在马铃薯(S. tuberosum)上。从12株带有黑色水渍状病斑且在叶背面有白色孢子形成的植株上采集了15片叶子,并在3天内进行处理。将叶子置于湿度箱中,在室温(平均温度18°C)黑暗条件下培养,直至观察到孢子形成。微观特征与疫霉属一致。通过在黑麦B琼脂平板上连续继代培养,仅获得了一个纯培养物。培养8天后,用蒸馏水冲洗平板,观察到卵形、半乳头状的棒状孢子囊,长38至41μm(平均39;N = 86),宽23至29μm(平均26;N = 86)。为了验证科赫法则并测试该分离株感染马铃薯以及哥伦比亚与马铃薯作物相关的茄属植物的能力,对刺茄、马铃薯和美洲茄(S. americanum,美洲龙葵)的三片离体叶片进行了三次重复致病性试验。使用血细胞计数器估计,从在黑麦B琼脂上生长8天的培养物中获得致病疫霉分离株的1×10孢子囊/ml悬浮液。将悬浮液在4°C下孵育2小时以诱导游动孢子释放。然后通过喷雾接种叶片直至径流。在湿度箱中于18°C培养5天后,在所有三个寄主的叶片中均观察到菌丝体、孢子囊和褐色病斑,类似于上述描述,表明具有致病性。从致病疫霉分离株中提取DNA(4)。扩增并测序了四个核基因座,即ITS、β-微管蛋白、Ras和Avr3a,以及一个线粒体基因,细胞色素c氧化酶1(Cox1)。使用Blastn将序列与GenBank数据库进行比较。在所有情况下,最佳匹配均对应致病疫霉(Avr3A的GenBank登录号为HQ639930,β-微管蛋白的为HQ639931,Cox1的为HQ639932,iRas的为HQ639933,Ras的为HQ639934,ITS的为JF419363)。致病疫霉在野生茄科植物上引起典型晚疫病症状的报道越来越频繁,其他国家如秘鲁也有此类报道(1)。据我们所知,这是首次在哥伦比亚从刺茄中观察到并分离出致病疫霉,这表明这种野生茄科杂草有可能成为晚疫病的另一个寄主。参考文献:(1)G. Garry等人,《欧洲植物病理学报》113:71,2005年。(2)R. Levin等人,《美国植物学杂志》92:603,2005年。(3)M. Nee,《茄属刺萼茄组的修订》,博士论文,威斯康星大学麦迪逊分校,1979年。(4)A. M. Vargas等人,《植物病理学》99:82,2009年。