Alippi A M, López A C
CIDEFI (CIC - CONICET), Facultad de Ciencias Agrarias y Forestales, UNLP, cc 31, 1900 La Plata, Argentina.
Plant Dis. 2009 Aug;93(8):842. doi: 10.1094/PDIS-93-8-0842C.
Peace lily (Spathiphyllum wallisii Regel) is a popular ornamental potted plant in Argentina. During May of 2008 (austral autumn), necrotic lesions of unknown etiology were observed on S. wallisii in a nursery in Pontevedra (34°45'6″S, 58°42'42″W). Plants first showed water-soaked areas starting from the leaf tips. Infected tissue became irregular, brown, dark-to-black lesions on leaves ~12 to 14 mm in diameter surrounded by yellowish haloes. Disease incidence approached 30%. Abundant bacterial streaming was observed from lesions when examined at ×100. Bacteria isolated from lesions formed white-to-cream, glistening, convex colonies on yeast dextrose calcium carbonate agar. Three bacterial strains isolated from different symptomatic plants were selected for comparative analysis with Pectobacterium carotovorum subsp. carotovorum type strain ATCC 15713. All were facultatively, anaerobic, gram-negative rods, pectolytic on crystal violet pectate agar, nonfluorescent on King's medium B, and elicited a hypersensitive response in tobacco plants. All strains were oxidase and arginine dihydrolase negative, fermented glucose, did not hydrolyze starch, did not produce lecithinase, indole or the blue pigment indigoidine, reduced nitrates, hydrolyzed gelatin and esculin, able to rot onion slices, caused soft rot of potato tubers, resistant to erythromycin, and grew at 37°C. Acid was produced from cellobiose, d-glucose, d-melibiose, d-mannitol, d-mannose, l-rhamnose, d-sucrose, and l-arabinose but not from inositol and d-sorbitol. Bacteria utilized N-acetyl-glucosamine and citrate but not tartrate, benzoate, or propionate. Their identity was confirmed by 16S rRNA gene sequencing of strain F402Pcc (GenBank Accession No. FJ717337) showing a 99% homology with that of strain ATCC 3326 (FJ 5958691). Pathogenicity was verified on S. wallisii, Dieffenbachia picta, Aglaonema commutatum, and Anthurium andraeanum within the Araceae family by spraying two plants per strain tested with bacterial suspensions (10 CFU/ml) in sterile distilled water with and without wounding the leaves with sterile needles. Controls were sprayed with sterile distilled water. After 48 h in a humidity chamber, inoculated plants and controls were maintained at 25 ± 3°C in a greenhouse. Water-soaked areas developed from 24 to 48 h after inoculation and became necrotic within 4 to 5 days. Lesions expanded to resemble natural infection in S. wallisii within 20 days, while in the rest of the hosts tested, lesions were smaller and remained brown surrounded by yellowish haloes. All strains were reisolated from each host tested. The original and all reisolated strains were compared by enterobacterial repetitive intergeneric consensus-PCR (4) confirming that DNA fingerprints of the reisolated strains were identical to those of the original strains. No lesions were observed on controls. The pathogen was identified as P. carotovorum subsp. carotovorum based on biochemical, physiological, pathogenicity tests, and 16S rRNA sequencing (1-3).To our knowledge, this is the first report of this pathogen on S. wallisii in Argentina although it has been reported as causing tomato pith necrosis (1) and soft rot of vegetables after harvest (3). References: (1) A. M. Alippi et al. Plant Dis. 81:230, 1997. (2) L. Gardan et al. Int. J. Syst. Evol. Microbiol. 53:381, 2003. (3) L. Halperin and L. S. Spaini. Rev. Argent. Agron. 6:261, 1939. (4) F. J. Louws et al. Appl. Environ. Microbiol. 60:2286, 1994.
白鹤芋(Spathiphyllum wallisii Regel)是阿根廷一种受欢迎的观赏盆栽植物。2008年5月(南半球秋季),在蓬特韦德拉(南纬34°45'6″,西经58°42'42″)的一个苗圃中,观察到白鹤芋出现了病因不明的坏死病变。植株最初在叶尖处出现水渍状区域。受感染组织在叶片上变成不规则的褐色、深至黑色病斑,直径约12至14毫米,周围有淡黄色晕圈。发病率接近30%。在100倍放大倍数下检查病斑时,观察到大量细菌溢菌现象。从病斑分离出的细菌在酵母葡萄糖碳酸钙琼脂上形成白色至奶油色、有光泽、凸起的菌落。从不同症状植株分离出的三株细菌菌株被选出来与胡萝卜软腐果胶杆菌胡萝卜软腐亚种模式菌株ATCC 15713进行比较分析。所有菌株都是兼性厌氧菌,革兰氏阴性杆菌,在结晶紫果胶酸盐琼脂上具有果胶分解能力,在金氏培养基B上不产生荧光,并且在烟草植株中引发过敏反应。所有菌株氧化酶和精氨酸双水解酶均为阴性,能发酵葡萄糖,不水解淀粉,不产生卵磷脂酶、吲哚或蓝色色素靛蓝素,能还原硝酸盐,水解明胶和七叶苷,能够使洋葱片腐烂,引起马铃薯块茎软腐,对红霉素有抗性,并且能在37°C下生长。能利用纤维二糖、d -葡萄糖、d -蜜二糖、d -甘露醇、d -甘露糖、l -鼠李糖、d -蔗糖和l -阿拉伯糖产酸,但不能利用肌醇和d -山梨醇产酸。细菌利用N -乙酰葡糖胺和柠檬酸盐,但不利用酒石酸盐、苯甲酸盐或丙酸盐。通过对菌株F402Pcc的16S rRNA基因测序(GenBank登录号FJ717337)确认其身份,显示与菌株ATCC 3326(FJ 5958691)有99%的同源性。通过用细菌悬浮液(10 CFU/ml)在无菌蒸馏水中对每个测试菌株的两株植物进行喷雾接种(有无用无菌针损伤叶片),并以喷雾无菌蒸馏水作为对照,在天南星科的白鹤芋、花叶万年青、粗肋草和红掌植株上验证了致病性。在湿度箱中放置48小时后,将接种的植株和对照置于温室中25±3°C条件下。接种后24至48小时出现水渍状区域,4至5天内变成坏死。20天内,白鹤芋病斑扩展至类似自然感染,而在其余测试宿主中,病斑较小,仍为褐色,周围有淡黄色晕圈。从每个测试宿主中都重新分离出了所有菌株。通过肠杆菌重复基因间共有序列PCR(4)对原始菌株和所有重新分离出的菌株进行比较,确认重新分离出的菌株的DNA指纹与原始菌株相同。对照植株未观察到病斑。根据生化、生理、致病性测试以及16S rRNA测序(1 - 3),该病原菌被鉴定为胡萝卜软腐果胶杆菌胡萝卜软腐亚种。据我们所知,这是该病原菌在阿根廷白鹤芋上的首次报道,尽管它曾被报道可引起番茄髓坏死(1)和收获后蔬菜软腐(3)。参考文献:(1)A. M. Alippi等人,《植物病害》81:230,1997年。(2)L. Gardan等人,《国际系统与进化微生物学杂志》53:381,2003年。(3)L. Halperin和L. S. Spaini,《阿根廷农业学报》6:261,1939年。(4)F. J. Louws等人,《应用与环境微生物学》60:2286,1994年。