Cantore P Lo, Iacobellis N S
Dipartimento di Biologia, Difesa e Biotecnologie Agro-Forestali, Università degli Studi della Basilicata, Viale dell'Ateneo Lucano 10, 85100 Potenza, Italy.
Plant Dis. 2007 May;91(5):638. doi: 10.1094/PDIS-91-5-0638A.
During the winter of 2004, symptoms were observed in commercial cauliflower (Brassica oleracea L. convar. botrytis (L.) Alef. var. italica) fields of "romanesco" type (cv. Navona) in Apulia, southern Italy. These symptoms were noted on inflorescences that were almost ready for harvest, and a bacterial etiology was suspected. In particular, the corymbs showed water-soaked and brown discolored areas which then rotted. The above alterations involved the whole inflorescences, or in some cases, only a few florets. Longitudinal sections of the symptomatic inflorescences or the single floret showed brown discoloration and rotting of the internal tissues. The disease caused severe crop losses (approximately 100% either in the field or after harvest). Bacteria were isolated from water-soaked and soft-rotted cauliflower heads on King's medium B (KB). The strains were purified on nutrient agar and assayed for pathogenicity on subcorymbs from freshly harvested cauliflower heads. Bacterial suspensions containing approximately 10 CFU/ml were then sprayed on the surface of subcorymbs (3 subcorymbs per strain). Furthermore, in other pathogenicity assays, the florets were dipped in 10 CFU/ml bacterial suspensions or small aliquots of inoculum were injected into the peduncle of subcorymbs with a sterile syringe. Cauliflower heads treated with sterile distilled water were used as controls. After inoculation, the subcorymbs were maintained at 25°C and approximately 100% relative humidity for 48 h. All bacterial strains either applied to cauliflower subcorymbs by spray inoculation or dipping reproduced the disease symptoms. Intensity of symptoms varied with the inoculation method. Injection of bacteria caused water soaking and soft rot of cauliflower internal tissues. No symptoms were observed in negative control subcorymbs inoculated with sterile water. All bacterial strains were gram negative and fluorescent on KB. Isolates (17 of 18) showed the LOPAT characters of group Vb (++-+-) fluorescent pseudomonads, and only strain USB1237 showed characters of group IVb (-+++-) (3). The pectolytic activity of the latter strain was confirmed by the pectinase plate assay (4). The identity of representative strains was confirmed by the nutritional profile obtained with the Biolog Identification System (MicroLogTM System Release 4.2; Biolog, Inc., Hayward, CA). Strains USB1224, USB1226, USB1228, USB1231, USB1235, USB1236, USB1238, and USB1239 were identified as Pseudomonas fluorescens with similarity indices of 0.86, 0.52, 0.73, 0.81, 0.73, 0.74, 0.69, and 0.85, respectively. The pectolytic strain USB1237 was identified as a Pseudomonas spp. that is closely related to P. putida (similarity index = 0.45). In conclusion, the above results indicate that P. fluorescens is responsible for head rot of cauliflower. A similar disease has been previously reported on broccoli in different areas (1,2), but to our knowledge, this is the first report of head rot of cauliflower caused by P. fluorescens. References: (1) C. H. Canaday et al. Phytopathology 77:1712, 1987. (2) P. D. Hildebrand. Can. J. Plant Pathol. 8:350, 1986. (3) R. A. Lelliott and D. E. Stead. Methods for the diagnosis of bacterial diseases of plants. In: Methods in Plant Pathology. Vol. 2, T. F. Preece, ed. Blackwell Scientific Publications, Oxford, UK, 1987. (4) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. The American Phytopathological Society, St. Paul, MN, 2001.
2004年冬季,在意大利南部普利亚大区的“罗马花椰菜”(品种名:纳沃纳)商业花椰菜(甘蓝变种,意大利花椰菜变种)田中观察到一些症状。这些症状出现在即将收获的花序上,怀疑是由细菌引起的。特别是,花球出现水渍状和褐色变色区域,随后腐烂。上述病变累及整个花序,在某些情况下,仅影响少数小花。有症状的花序或单个小花的纵切面显示内部组织褐色变色和腐烂。这种病害导致了严重的作物损失(田间或收获后损失率约为100%)。从水渍状和软腐的花椰菜头中在金氏B培养基(KB)上分离出细菌。将菌株在营养琼脂上纯化,并在新鲜收获的花椰菜头的小分枝上测定致病性。然后将含有约10CFU/ml的细菌悬液喷洒在小分枝表面(每个菌株3个小分枝)。此外,在其他致病性测定中,将小花浸入10CFU/ml细菌悬液中,或用无菌注射器将少量接种物注入小分枝的花梗中。用无菌蒸馏水处理的花椰菜头用作对照。接种后,将小分枝在25°C和相对湿度约100%的条件下保持48小时。通过喷雾接种或浸泡应用于花椰菜小分枝的所有细菌菌株都再现了病害症状。症状的严重程度因接种方法而异。注射细菌导致花椰菜内部组织水渍状和软腐。用无菌水接种的阴性对照小分枝未观察到症状。所有细菌菌株在KB上革兰氏阴性且发荧光。分离株(18个中的17个)表现出Vb组(++-+-)荧光假单胞菌的LOPAT特征,只有USB1237菌株表现出IVb组(-+++-)的特征(3)。通过果胶酶平板测定法(4)证实了后一种菌株的果胶分解活性。用Biolog鉴定系统(MicroLogTM系统版本4.2;Biolog公司,加利福尼亚州海沃德)获得的营养图谱证实了代表性菌株的身份。USB1224、USB1226、USB1228、USB1231、USB1235、USB1236、USB1238和USB1239菌株被鉴定为荧光假单胞菌,相似性指数分别为0.86、0.52、0.73、0.81、