Khoodoo M H R, Ganoo E S, Saumtally S
Plant Pathology Department, Mauritius Sugar Industry Research Institute, Réduit, Mauritius.
Plant Dis. 2007 Sep;91(9):1200. doi: 10.1094/PDIS-91-9-1200B.
In October 2005 and September 2006, two outbreaks of bacterial wilt occurred in the south and north (90 and 95 m above sea level, respectively) of Mauritius, respectively, on different potato cultivars in seed potato fields. Symptoms were reported at harvest when profuse creamy exudates were observed oozing from the eyes of tubers. The brown appearance of the vascular rings, which was accompanied by extensive maceration, suggested potato brown rot. Severe symptoms with complete rotting of vascular tissues and oozing from heel ends of tubers were commonly observed. Ralstonia solanacearum has been regularly encountered for decades around the island, but before October 2005, all isolates belonged to Race 1 biovar 3. The pathogen was isolated from samples collected from the two outbreaks by plating on Kelman's medium amended with 100 ppm of polymixin B sulfate. Thirty-three isolates were obtained from stems and tubers of potato cvs. Spunta, Delaware, Atlantic, and Belle Isle, from soil samples, and weed hosts Solanum americanum, Lycopersicon pimpinellifolium, and Oxalis latifolia. These weeds, however, did not show symptoms of wilting or vascular browning, although oozing was observed when the stems were cut and placed in water. When reinoculated in tomato bioassays, 17 tested isolates caused wilting and were successfully reisolated, confirming Koch's postulates. All colonies were positive for Ralstonia by the Spot√Check LF test (Adgen, Ayr, UK) and by indirect plate-trapped antigen-ELISA (Agden) using monoclonal antibodies raised against Race 3 strains. Isolate biovar was determined by performing standard biochemical tests (1). All 33 isolates metabolized maltose, lactose, and cellobiose but not trehalose and the hexose alcohols dulcitol, mannitol, and sorbitol, thereby showing that they all belong to biovar 2 of Andean phenotype 2A. The final identification was performed by a PCR test using Race 1 specific primers PSIF and PSIR (4) and Race 3 specific primers 630 and 631 (3). The Race 3 specific band was amplified from all isolates while the Race 1 specific band was not. Assignment to biovar 2 was independently confirmed by CABI Identification Service, UK. R. solanacearum R3bv2 is distributed worldwide, occurring in temperate regions, subtropical areas, and at higher altitudes in the tropics, reportedly because of its lower temperature optimum. Brown rot is often disseminated by seed potato tubers that are latently infected by the pathogen (2). Seed potato fields typically undergo a 7-year crop rotation with sugar cane in Mauritius, so it is unlikely that the pathogen was present in these fields for a long time. The infection of weeds in these same fields was probably due to the movement of water contaminated by tuber exudates. Epidemiological results suggest that R. solanacearum R3bv 2A was recently introduced into Mauritius, although its origin is not known. Generally, R3bv2 strains around the world appear to be clonal and seem to be spreading rapidly into previously uninfested areas such as Mauritius. Stronger standards for seed potato testing may be needed to prevent a wide dissemination of R3bv2. References: (1) A. C. Hayward. J. Appl. Bacteriol. 27:265, 1964. (2) A. C. Hayward et al. Page 420 in: Bacterial Wilt Disease: Molecular and Ecological Aspects. P. Prior et al., eds. Springer, Berlin, 1998. (3) M. Fegan et al. Page 19 in: Bacterial Wilt Disease: Molecular and Ecological Aspects. P. Prior et al., eds. Springer, Berlin, 1998. (4) Y.-A. Lee et al. Appl. Environ. Microbiol. 67:3943, 2001.
2005年10月和2006年9月,毛里求斯南部和北部(分别海拔90米和95米)的种薯田不同马铃薯品种上分别发生了两起青枯病疫情。收获时发现症状,当时观察到大量乳状渗出物从块茎芽眼渗出。维管束环呈现褐色,伴有广泛的软腐,提示为马铃薯褐腐病。通常观察到严重症状,维管束组织完全腐烂,块茎根部渗出。几十年来,该岛周围经常发现青枯雷尔氏菌,但在2005年10月之前,所有分离株都属于1号小种3型生化变种。通过接种于添加了100 ppm硫酸多粘菌素B的凯尔曼培养基上,从两次疫情采集的样本中分离出病原菌。从马铃薯品种Spunta、Delaware、Atlantic和Belle Isle的茎和块茎、土壤样本以及杂草寄主美洲茄、醋栗番茄和阔叶酢浆草中获得了33个分离株。然而,这些杂草没有出现萎蔫或维管束褐变症状,尽管将茎切断并置于水中时观察到有渗出。在番茄生物测定中再次接种时,17个受试分离株引起萎蔫,并成功重新分离,证实了科赫法则。通过斑点检测LF试验(英国艾尔市Adgen公司)以及使用针对3号小种菌株产生的单克隆抗体的间接平板捕获抗原ELISA(Adgen公司),所有菌落对青枯雷尔氏菌均呈阳性。通过进行标准生化试验确定分离株的生化变种(1)。所有33个分离株都能代谢麦芽糖、乳糖和纤维二糖,但不能代谢海藻糖以及己糖醇卫矛醇、甘露醇和山梨醇,因此表明它们都属于安第斯表型2A的2型生化变种。通过使用1号小种特异性引物PSIF和PSIR(4)以及3号小种特异性引物630和631(3)进行PCR试验进行最终鉴定。从所有分离株中扩增出了3号小种特异性条带,而1号小种特异性条带未扩增出。英国国际农业与生物科学中心鉴定服务部独立确认其属于2型生化变种。青枯雷尔氏菌R3bv2分布于全球,存在于温带地区、亚热带地区以及热带地区的较高海拔处,据报道是因为其最适温度较低。褐腐病常通过被病原菌潜伏感染的种薯块茎传播(2)。在毛里求斯,种薯田通常与甘蔗进行7年轮作,因此该病原菌在这些田地中不太可能长期存在。这些相同田地中杂草的感染可能是由于被块茎渗出物污染的水的流动所致。流行病学结果表明,青枯雷尔氏菌R3bv 2A最近被引入毛里求斯,尽管其来源不明。一般来说,世界各地的R3bv2菌株似乎是克隆性的,并且似乎正在迅速传播到以前未受侵染的地区,如毛里求斯。可能需要更严格的种薯检测标准以防止R3bv2的广泛传播。参考文献:(1)A. C. 海沃德。《应用细菌学杂志》27:265,1964年。(2)A. C. 海沃德等人。载于:《青枯病:分子与生态方面》。P. 普里尔等人编。施普林格出版社,柏林,1998年,第420页。(3)M. 费根等人。载于:《青枯病:分子与生态方面》。P. 普里尔等人编。施普林格出版社,柏林,1998年,第19页。(4)Y.-A. 李等人。《应用与环境微生物学》67:3943,2001年。