Penyalver R, Sánchez J J, Petit A, Salcedo C I, López M M
Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Apartado oficial, 46113 Moncada, Valencia, Spain.
Institut des Sciences Végétales, Centre National de Recherche Scientifique, 91198 Gif-sur-Yvette, France.
Plant Dis. 2004 Apr;88(4):428. doi: 10.1094/PDIS.2004.88.4.428C.
Agrobacterium-like colonies were recovered onto Roy-Sasser's medium from a young tumor (4 cm in diameter) on the stem of weeping fig (Ficus benjamina L.), 10 cm from the crown. The galled plant was collected in 1999 from a garden center in Valencia, Spain. After colony purification and tomato and weeping fig plant inoculations, one nonpathogenic and five Agrobacterium isolates that were tumorigenic in both plant species were characterized. On the basis of biovar classification tests, the nonpathogenic isolate was identified as belonging to biovar 1 of Agrobacterium (now called A. tumefaciens), whereas the tumorigenic isolates could not be assigned to any of the known Agrobacterium biovars. The isolates were positive for oxidase, growth in 2% NaCl, production of alkali from l-tartaric acid, and production of acid from mannitol-CaCO and negative for 3-ketolactose production, growth and pigmentation in ferric ammonium citrate, growth at 35°C, citrate utilization, acid production from sucrose and melezitose, and alkali production from malonic acid. Nopaline was the unique opine found in galls induced in weeping fig plants inoculated with the pathogenic isolates. Moreover, all isolates utilize the opine nopaline, but not octopine, manopine, agropine, chrysopine, cucumopine, or mikimopine. They were susceptible to agrocin 84 produced by strain K84. Heat-treated bacterial suspensions of these isolates yielded the expected amplification product using polymerase chain reaction (PCR) with the FGPtmr530/FGPtmr701' primers pair from the tmr gene (3). Aerial gall disease was first reported on F. benjamina in Florida (1), and the isolated agrobacteria belongs to a new species named A. larrymoorei (2). Later, tumorigenic agrobacteria from weeping fig galls were isolated in Italy and the Netherlands (4). Our data suggest that the tumorigenic strains isolated in Spain differ greatly from those first described in the United States (1) on the basis of alkali production from l-tartaric acid, chrysopine detection on tumors, susceptibility to agrocin 84, and tmr amplification, but they might be similar to some of the Italian isolates (4). To our knowledge, this is the first report of isolation of tumorigenic Agrobacterium sp. from F. benjamina L. in Spain. References: (1) H. Bouzar et al. Appl. Environ. Microbiol. 61:65, 1995. (2) H. Bouzar and J. B. Jones. Int. J. Syst. Evol. Microbiol. 51:1023. 2001. (3) X. Nesme et al. Pages 47-50 in: Endocytobiology IV. P. Nardon et al. eds. INRA, France, 1989. (4) A. Zoina et al. Plant Pathol. 50:620, 2001.
从垂叶榕（Ficus benjamina L.）茎干上一个直径4厘米、距树冠10厘米的幼嫩肿瘤中，将类似根癌土壤杆菌的菌落转接至罗伊 - 萨瑟培养基上。该患瘤植物于1999年从西班牙巴伦西亚的一个园艺中心采集。经过菌落纯化以及对番茄和垂叶榕植株的接种后，对一株非致病性菌株和五株在两种植物中均具有致瘤性的根癌土壤杆菌分离株进行了特性鉴定。根据生物型分类测试，非致病性分离株被鉴定为属于根癌土壤杆菌生物型1（现称为根癌农杆菌），而致瘤性分离株无法归入任何已知的根癌土壤杆菌生物型。这些分离株氧化酶试验呈阳性，能在2%氯化钠中生长，能由L - 酒石酸产碱，能由甘露醇 - 碳酸钙产酸，3 - 酮乳糖产生试验、在柠檬酸铁铵中生长及色素形成试验、35°C生长试验、柠檬酸盐利用试验、由蔗糖和松三糖产酸试验以及由丙二酸产碱试验均呈阴性。胭脂碱是在用致病性分离株接种的垂叶榕植株中诱导形成的瘿瘤中发现的唯一冠瘿碱。此外，所有分离株都利用胭脂碱，但不利用章鱼碱、甘露碱、农杆碱、金雀花碱、黄瓜碱或米基莫碱。它们对K84菌株产生的农杆菌素84敏感。使用来自tmr基因的FGPtmr530/FGPtmr701'引物对通过聚合酶链反应（PCR）对这些分离株的热处理细菌悬液进行扩增，得到了预期的扩增产物。气生瘿瘤病首次在佛罗里达州的垂叶榕上被报道（1），分离出的土壤杆菌属于一个名为拉里摩尔根癌农杆菌的新物种（2）。后来，在意大利和荷兰从垂叶榕瘿瘤中分离出了致瘤性土壤杆菌（4）。我们的数据表明，在西班牙分离出的致瘤菌株在由L - 酒石酸产碱、肿瘤中金雀花碱的检测、对农杆菌素84的敏感性以及tmr扩增方面与在美国首次描述的菌株（1）有很大差异，但它们可能与一些意大利分离株（4）相似。据我们所知，这是在西班牙从垂叶榕中分离出致瘤性根癌土壤杆菌属的首次报道。参考文献：（1）H. Bouzar等人，《应用与环境微生物学》61:65，1995年。（2）H. Bouzar和J. B. Jones，《国际系统与进化微生物学杂志》51:1023，2001年。（3）X. Nesme等人，载于《内共生生物学IV》第47 - 50页，P. Nardon等人编，法国国家农业研究院，1989年。（4）A. Zoina等人，《植物病理学》50:620，2001年。
