Sikirou R, Beed F, Ezin V, Gbèhounou G, Miller S A, Wydra K
Institut National des Recherches Agricoles du Benin/Laboratoire de Défense des Cultures, 01 B.P. 128 Porto-Novo, Benin.
International Institute of Tropical Agriculture (IITA), Plot 15 Naguru, Kampala, Uganda.
Plant Dis. 2009 May;93(5):549. doi: 10.1094/PDIS-93-5-0549B.
In June 2004, wilted tomatoes with no foliar yellowing were observed in Ouègbo, Atlantique District, Benin. The cut tomato stems released whitish bacterial ooze. Longitudinal sections of most stems showed brown vascular discoloration. Twenty symptomatic tomato plants were collected from 10 fields and exported to the Institute of Plant Disease and Plant Protection, Leibniz Universität Hannover, Germany. Bacteria were isolated on triphenyl tetrazolium chloride (TTC) medium (2) and three of the nine bacterial isolates that resembled Ralstonia solanacearum (colonies with red center and whitish periphery) and reference strain ToUdk (race 1 biovar 3; N. Thaveechai, Kasetsart University, Bangkok, Thailand) were used for pathogenicity tests. Five 4-week-old tomato plants cv. Tohounvi, grown in individual plastic pots (14 × 16 cm) containing sterilized field soil, were inoculated with each of the four isolates individually by soil drenching with 30 ml of the test cultures at 10 CFU/ml. Control plants were treated with 30 ml of sterile water. All plants were incubated in a glasshouse at 30°C. All plants inoculated with the isolates from Benin wilted 4 days after inoculation with symptoms similar to those observed in the field. Plants inoculated with the reference strain wilted 7 to 11 days after inoculation. Control plants treated with water remained healthy. R. solanacearum was recovered from the 20 symptomatic plants on TTC medium. The identity of the strains in comparison with the reference strain was confirmed by PCR with species-specific primers 759/760, which produced a single 281-bp fragment (3). Because similar symptoms were being increasingly reported by farmers across Benin and linked with reduced tomato yields, a disease survey was undertaken by IITA in 2006 and 2007. Wilted tomato plants were found across all agro-ecological zones of Benin (3 to 72% of plants per field). Isolates were recovered from the southeastern districts of Adja-Ouèrè, Sakété, Adjohoun, and Dangbo, the southwestern districts of Klouékanmè and Athiémé, the southern districts of Toffo and Bohicon, the central districts of Dassa and Savè, and the northern districts of Malanville and Karimama. Identification of R. solanacearum was confirmed following inoculation of tomato, production of characteristic wilting symptoms, recovery of the pathogen on TTC medium, and positive identification with ELISA kits (Pathoscreen Rs; Agdia Inc., Elkhart, IN). To our knowledge, this is the first report of R. solanacearum infecting tomato in Benin. Tomato is the most cultivated vegetable crop in Benin and important to the livelihood of many people in peri-urban and rural areas. Understanding that the cause of the observed crop losses is R. solanacearum may lead to implementation of management strategies such as deployment of disease-resistant cultivars or grafting tomatoes onto bacterial wilt-resistant rootstocks (1). References: (1) P. Aggarwal et al. Indian J. Agric. Sci. 78:379, 2008. (2) A. Kelman. Phytopathology 44:693, 1954. (3) N. Opina et al. Asian Pac. J. Mol. Biol. Biotechnol. 5:19, 1997.
2004年6月,在贝宁大西洋省韦格博地区发现了一些番茄植株枯萎,但叶片未发黄。切开的番茄茎部会渗出白色菌脓。大多数茎的纵切面显示出褐色的维管束变色。从10个田地中采集了20株有症状的番茄植株,并运往德国汉诺威莱布尼茨大学植物病害与植物保护研究所。细菌在氯化三苯基四氮唑(TTC)培养基上分离培养(2),9株与青枯雷尔氏菌相似的细菌分离物(菌落中心呈红色,边缘发白)中的3株以及参考菌株ToUdk(1号小种3型;泰国曼谷 Kasetsart 大学的 N. Thaveechai)用于致病性测试。将5株4周龄的番茄品种Tohounvi种植在装有灭菌田间土壤的单个塑料盆(14×16厘米)中,分别用30毫升浓度为10 CFU/ml的测试培养物对每株植物进行灌根接种这4种分离物。对照植株用30毫升无菌水处理。所有植株在温室中于30°C下培养。所有接种了来自贝宁分离物的植株在接种后4天枯萎,症状与在田间观察到的相似。接种参考菌株的植株在接种后7至11天枯萎。用水处理的对照植株保持健康。在TTC培养基上从20株有症状的植株中分离出了青枯雷尔氏菌。通过使用种特异性引物759/760进行PCR,与参考菌株相比,确认了菌株的身份,该引物产生了一个单一的281-bp片段(3)。由于贝宁各地的农民越来越多地报告出现类似症状,并与番茄产量下降有关,国际热带农业研究所(IITA)在2006年和2007年进行了一项病害调查。在贝宁所有农业生态区都发现了枯萎的番茄植株(每个田地中植株的3%至72%)。从东南部的阿贾-韦雷、萨凯特、阿乔洪和当博地区,西南部的克洛埃卡姆和阿蒂埃梅地区,南部的托福和博希孔地区,中部的达萨和萨韦地区以及北部的马兰维尔和卡里马马地区分离出了菌株。通过接种番茄、产生典型的萎蔫症状、在TTC培养基上分离出病原体以及使用ELISA试剂盒(Pathoscreen Rs;Agdia公司,美国印第安纳州埃尔克哈特)进行阳性鉴定,确认了青枯雷尔氏菌的身份。据我们所知,这是青枯雷尔氏菌在贝宁感染番茄的首次报道。番茄是贝宁种植最多的蔬菜作物,对许多城市周边和农村地区的人们的生计至关重要。了解到观察到的作物损失的原因是青枯雷尔氏菌,可能会导致实施一些管理策略,如部署抗病品种或将番茄嫁接到抗青枯病的砧木上(1)。参考文献:(1)P. Aggarwal等人,《印度农业科学杂志》78:379,2008年。(2)A. Kelman,《植物病理学》44:693,1954年。(3)N. Opina等人,《亚太分子生物学与生物技术杂志》5:19,1997年。