Thera A T, Jacobsen B J, Neher O T
Programme Fruits et Legumes, Institut d'Economie Rurale, BP 258, Rue, Mohamed V, Bamako, Republique du Mali.
Department of Plant Sciences and Plant Pathology, Montana State University, 119 Plant Biosciences Building, Bozeman 59717-3150.
Plant Dis. 2010 Mar;94(3):372. doi: 10.1094/PDIS-94-3-0372B.
Ralstonia solanacearum (Smith) Yabuuchi et al. causes bacterial wilt worldwide on a wide range of plant species. In Mali, the disease is commonly found on potato (Solanum tuberosum L.), tomato (Lycopersicon esculentum var. esculentum L.), pepper (Capsicum annuum L.), eggplant (Solanum melongena L.), tobacco (Nicotiana tabacum L.), and peanut (Arachis hypogaea L.). Determination of race and biovar is critical for development of potato seed certification programs for management of the disease. Isolates (25) of R. solanacearum were obtained from wilting potato, pepper, eggplant, tobacco, and tomato plants collected from fields near Baguineda, Sonityeni, Sotuba, Sikasso, and Kolikoro. Isolations were made from bacterial streaming by dilution plating on triphenyl tetrazolium chloride medium (TZC) (2). Characteristic colonies were selected and identified by ELISA or Immunostrips (Pathoscreen Rs, Agdia Inc., Elkhart, IN). These isolates were used in host range studies and hypersensitivity (HR) tests on tobacco (cv. xanthi) (3) and tested for their ability to produce acids on Ayers basal media amended with disaccharide and hexose alcohol carbon sources (1). These isolates caused characteristic wilt 40 days postinoculation on greenhouse-grown tobacco (cv. Xanthi), peanut (cv. 4610), and tomato (cv. Roma VF) plants when stems of five plants of each host were syringe inoculated with 0.1 ml of a 1 × 10 CFU/ml of bacteria. Plants inoculated with sterile distilled water remained symptomless and R. solanacearum was reisolated from infected plants on TZC and identified with Immunostrips. All HR tests were negative. Infection of peanut, tobacco, and tomato and the results of the HR tests indicated that all isolates were Race 1 and no significant variation was noted between isolates. Acid was produced from the hexose alcohols: mannitol, sorbitol, and dulcitol; and the disaccharides: cellobiose, lactose, and maltose. This indicated that all isolates were biovar 3, the same as a known Race 1 strain from tobacco (MSU Plant Pathology teaching collection) (1). To assess relative distribution of R. solanacearum, 20 soil samples collected from potato fields in the vicinity of Baguineda, Kati, Koulikoro, and Sikasso were placed in pots (30 × 25 cm) under shade cloth at the IER Station in Sotuba and planted with 30-day-old tobacco plants. After 90 days, infected plants (35 to 100% infection) were found in all soils. Infected plants exhibited classical wilt symptoms and tested positive for R. solanacearum infections as confirmed by Immunostrip tests. Six of nine surface water samples taken near potato fields in Baguineda, Sikasso, Mopti, and Koulikoro tested positive for the presence of R. solanacearum by an Agdia Inc. enrichment kit and ELISA. A weed, Commelina forskalaei (Vahl), collected by Farako creek near Sikasso tested positive in the Immunostrip test even though no symptoms were obvious. No attempt was made to characterize the race, biovar, or phylotype of the soil, water, and weed isolates. To our knowledge, this is the first time that the race and biovar of R. solanacearum from Mali has been reported and the wide distribution of this pathogen in Malian soils and surface water has been demonstrated. It is significant that we did not detect Race 3 biovar 2, which is subject to quarantine and biosecurity regulations. References: (1) A. C. Hayward. J. Bacteriol. 27:265, 1964. (2) A. Kelman. Phytopathology 44:693, 1954. (3) J. Lozano and L. Sequeira. Phytopathology 60:833, 1970.
青枯雷尔氏菌(Smith)Yabuuchi等人在全球范围内导致多种植物发生青枯病。在马里,该病常见于马铃薯(Solanum tuberosum L.)、番茄(Lycopersicon esculentum var. esculentum L.)、辣椒(Capsicum annuum L.)、茄子(Solanum melongena L.)、烟草(Nicotiana tabacum L.)和花生(Arachis hypogaea L.)上。确定小种和生化变种对于制定马铃薯种子认证计划以管理该病至关重要。从巴吉内达、索尼特尼、索图巴、锡卡索和科利科罗附近田间采集的萎蔫马铃薯、辣椒、茄子、烟草和番茄植株中获得了25株青枯雷尔氏菌分离株。通过在氯化三苯基四氮唑培养基(TZC)上稀释平板培养从细菌溢泌中进行分离(2)。挑选出特征性菌落,并通过酶联免疫吸附测定(ELISA)或免疫试纸条(Pathoscreen Rs,Agdia公司,印第安纳州埃尔克哈特)进行鉴定。这些分离株用于烟草(品种xanthi)的寄主范围研究和过敏反应(HR)测试(3),并测试它们在添加了二糖和己糖醇碳源的艾尔斯基础培养基上产酸的能力(1)。当用0.1 ml 1×10 CFU/ml的细菌通过注射器接种到每种寄主的5株植物的茎上时,这些分离株在接种后40天在温室种植烟草(品种Xanthi)、花生(品种4610)和番茄(品种Roma VF)植株上引起典型的萎蔫症状。接种无菌蒸馏水的植株无症状,并且在TZC上从感染植株中重新分离出青枯雷尔氏菌,并用免疫试纸条进行鉴定。所有HR测试均为阴性。花生、烟草和番茄的感染以及HR测试结果表明所有分离株均为小种1,分离株之间未观察到显著差异。从己糖醇:甘露醇、山梨醇和卫矛醇;以及二糖:纤维二糖、乳糖和麦芽糖中产酸。这表明所有分离株均为生化变种3,与来自烟草的已知小种1菌株(密歇根州立大学植物病理学教学收藏)相同(1)。为了评估青枯雷尔氏菌的相对分布,从巴吉内达、卡蒂、库利科罗和锡卡索附近的马铃薯田采集的20份土壤样品被放置在索图巴IER站的遮荫布下的花盆(30×25 cm)中,并种植30日龄的烟草植株。90天后,在所有土壤中均发现了感染植株(感染率为35%至100%)。感染植株表现出典型的萎蔫症状,并且通过免疫试纸条测试证实青枯雷尔氏菌感染呈阳性。在巴吉内达、锡卡索、莫普提和库利科罗附近的马铃薯田采集的9份地表水样品中有6份通过Agdia公司的富集试剂盒和ELISA检测出青枯雷尔氏菌呈阳性。在锡卡索附近的法拉科溪采集的一种杂草鸭跖草(Vahl),即使没有明显症状,在免疫试纸条测试中也呈阳性。未尝试对土壤、水和杂草分离株的小种、生化变种或系统型进行鉴定。据我们所知,这是首次报道来自马里的青枯雷尔氏菌的小种和生化变种,并且证明了该病原菌在马里土壤和地表水中的广泛分布。重要的是我们未检测到受检疫和生物安全法规管制的小种3生化变种2。参考文献:(1)A. C. Hayward。《细菌学杂志》27:265,1964年。(2)A. Kelman。《植物病理学》44:693,1954年。(3)J. Lozano和L. Sequeira。《植物病理学》60:833,1970年。