Bastas K K, Karakaya A
Selcuk University, Faculty of Agriculture, Department of Plant Protection, Konya, 42075, Turkey.
Ankara University, Faculty of Agriculture, Department of Plant Protection, Dıskapı, Ankara, 06110, Turkey.
Plant Dis. 2012 Mar;96(3):452. doi: 10.1094/PDIS-08-11-0675.
A new disease was observed during the spring and autumn of 2009 and 2010 on kiwifruit plants (Actinidia deliciosa cv. Hayward) in Rize Province of Turkey. Disease incidence was estimated as 3% in approximately 10 ha. Symptoms were characterized by dark brown spots surrounded by yellow halos on leaves and cankers with reddish exudate production on twigs and stems. Eight representative bacterial strains were isolated from leaf spots and tissues under the bark on King's B medium (KB) and identified as Pseudomonas syringae pv. actinidiae on the basis of biochemical, physiological (1,2), and PCR tests (3). Bacteria were gram negative, rod shaped, and nonfluorescent on KB; positive for levan production, sucrose and inositol utilization, and tobacco (Nicotiana tabacum cv. White Burley) hypersensitivity; and negative for growth at 37°C, oxidase, potato soft rot, arginine dihydrolase, urease, arbutin, erythritol, lactic acid, aesculin hydrolysis, gelatin liquefaction, and syringomycin production. Identity of the eight isolates was confirmed by PCR using P. syringae pv. actinidiae-specific primers PsaF1/R3 to generate a 280-bp DNA fragment (3). P. syringae pv. actinidiae reference strain NCPPB 3739, and CJW7 from Jae Sung Jung, Department of Biology, Sunchon National University, Korea, were employed in all biochemical, physiological, and molecular tests as positive controls. Pathogenicity was confirmed by artificial inoculation of 2-year-old A. deliciosa cv. Hayward. A bacterial suspension (10 CFU ml) was injected into kiwifruit twig tips, stems, and leaves with a hypodermic syringe, and the inoculated plants were placed at 25 to 28°C and 80% relative humidity growth chamber for 3 weeks. First symptoms were observed on leaves within 5 days after inoculation and on twigs after 20 days. No symptoms were observed on control plants that were inoculated with sterile water. Reisolation was made from dark brown lesions surrounded by yellow halos on leaves and cankers on twigs and stem and their identities were confirmed using the techniques previously described. All tests were performed three times and pathogenicity tests employed three plants for each strain. To our knowledge, this is the first report of P. syringae pv. actinidiae causing disease on kiwifruit in Turkey. Kiwifruit production in Turkey has expanded rapidly during the last 10 years ( http://www.tuik.gov.tr ) and phytosanitary measures are needed to prevent further spread of the bacterium to other kiwifruit orchards. References: (1) Y. J. Koh et al. N. Z. J. Crop Hortic. Sci. 38:4, 275, 2010. (2) R. A. Lelliott and D. E. Stead. Methods for the Diagnosis of Bacterial Diseases of Plants. Blackwell Scientific, Sussex, UK, 1988. (3) J. Rees-George et al. Plant Pathol. 59:453, 2010.
2009年春、秋两季以及2010年春、秋两季,在土耳其里泽省的猕猴桃植株(美味猕猴桃品种海沃德)上发现了一种新病害。在约10公顷的果园中,病害发生率估计为3%。症状表现为叶片上有暗褐色斑点,周围有黄色晕圈,嫩枝和茎上有溃疡,并产生带红色的渗出物。从叶片斑点和树皮下方组织中,在King氏B培养基(KB)上分离出8株具有代表性的细菌菌株,基于生化、生理(1,2)和PCR检测(3),鉴定为猕猴桃细菌性溃疡病菌。这些细菌革兰氏阴性,杆状,在KB培养基上不产生荧光;产果聚糖、利用蔗糖和肌醇、对烟草(白肋烟品种)过敏反应呈阳性;在37°C下不生长、氧化酶、马铃薯软腐、精氨酸双水解酶、脲酶、熊果苷、赤藓醇、乳酸、七叶苷水解、明胶液化和丁香霉素产生呈阴性。使用猕猴桃细菌性溃疡病菌特异性引物PsaF1/R3进行PCR,扩增出一个280 bp的DNA片段,从而确认了这8株分离菌的身份(3)。在所有生化、生理和分子检测中,均使用猕猴桃细菌性溃疡病菌参考菌株NCPPB 3739以及韩国顺天国立大学生物系Jae Sung Jung提供的CJW7作为阳性对照。通过对2年生美味猕猴桃品种海沃德进行人工接种来确认致病性。用皮下注射器将细菌悬浮液(10 CFU/ml)注入猕猴桃嫩梢顶端、茎和叶片,接种后的植株置于温度为25至28°C、相对湿度80%的生长室中培养3周。接种后5天内在叶片上观察到最初症状,20天后在嫩枝上观察到症状。接种无菌水的对照植株未出现症状。从叶片上被黄色晕圈包围的暗褐色病斑以及嫩枝和茎上的溃疡处重新分离细菌,并使用上述方法确认其身份。所有检测均重复进行3次,致病性检测中每个菌株使用3株植株。据我们所知,这是猕猴桃细菌性溃疡病菌在土耳其导致猕猴桃发病的首次报道。在过去10年中,土耳其的猕猴桃产量迅速增长(http://www.tuik.gov.tr),需要采取植物检疫措施以防止该细菌进一步传播到其他猕猴桃果园。参考文献:(1)Y. J. Koh等人,《新西兰作物与园艺科学杂志》38:4, 275, 2010。(2)R. A. Lelliott和D. E. Stead,《植物细菌病害诊断方法》,英国苏塞克斯布莱克韦尔科学出版社,1988。(3)J. Rees-George等人,《植物病理学》59:453, 2010。