Bastas K K, Sahin F
Selcuk University, Faculty of Agriculture, Department of Plant Protection, Campus, TR-42075 Konya, Turkey.
Yeditepe University, Faculty of Engineering and Architecture, Department of Genetics and Bioengineering TR-34755 Istanbul, Turkey.
Plant Dis. 2014 Jan;98(1):153. doi: 10.1094/PDIS-03-13-0220-PDN.
Fire blight, caused by Erwinia amylovora (Burr.) Winslow et al., affects plants in the Rosaceae family, which includes trees and shrubs in orchards, nurseries, and landscape plantations. During the springs and summers of 2008 and 2010, dying branches, necrotic leaves attached to shoots, and blighted twigs of meadowsweet (Spirea prunifolia) were observed at three different locations of landscape areas in Konya Province, Turkey. Disease incidence was approximately 1% on the plants during the surveys. Initial symptoms of reddish to brownish streaks on the shoots of infected plants were observed in spring. Nine representative bacterial strains were isolated from the lesions on shoots of seven meadowsweet plants on nutrient sucrose agar (NSA) medium and identified as E. amylovora on basis of biochemical, physiological (2,3) and molecular tests (1). Bacteria were gram-negative, rod shaped, aerobic, fermentative, yellow-orange on Miller and Scroth medium (2), positive for levan formation and acetoin production, did not grow at 36°C, positive for gelatin hydrolysis, and negative for esculin hydrolysis, indole, urease, catalase, oxidase, arginine dehydrolase, reduction of nitrate, acid production from lactose, and inositol. All strains were hypersensitive response-positive on tobacco (Nicotiana tabacum var. White Burley) plants. All strains were identified as E. amylovora using the species-specific primers set, A/B (1), by PCR assay, and by fatty acid methyl ester (FAME) profiles determined by Sherlock Microbial Identification System software (TSBA 6 v. 6.00; Microbial ID, Newark, DE) with similarity indices ranging from of 79 to 99%. Pathogenicity was tested by injecting of petioles and actively growing three shoot tips of 2-year-old S. prunifolia seedlings cv. number 29 using a 0.46 mm-diameter hypodermic needle with bacterial suspensions containing 10 CFU mL in sterile distilled water (SDW) Plants were inoculated with each of the nine bacterial strains and two references strains, Ea29 and NCPPB 2791 (Selcuk University, Department of Plant Protection, Konya, Turkey). Symptoms resembling those associated with natural infection appeared on the inoculated plants 7 days after inoculation. Plants inoculated with SDW served as a negative control treatment, and no symptoms were observed on these plants. All tests were repeated three times with the same results. Bacterial re-isolations were attempted from the control plants as well as shoots and leaves inoculated with the two reference strains and the nine bacteria identified as E. amylovora. Bacteria isolated from inoculated plants were identified as E. amylovora using the biochemical, physiological, and molecular tests described above, but this bacterium was not isolated from the control plants. Phytosanitary measures must be taken to avoid spread of the pathogen to ornamentals in new landscape areas in Turkey. This report is important because infected Spirea spp. can be a potential inoculum source for other rosaceous ornamentals. To our knowledge, this is the first report of the occurrence of fire blight on meadowsweet in Turkey. References: (1) S. Bereswill et al. Appl. Environ. Microbiol. 58:3522, 1992. (2) A. L. Jones and K. Geider. Laboratory Guide for Identification of Plant Pathogenic Bacteria, pp. 40-55. American Phytopathological Society, St. Paul, MN, 2001. (3) R. A. Lelliott and D. E. Stead. Methods for Diagnosis of Bacterial Diseases of Plants (Methods in Plant Pathology). Oxford, UK, 1987.
火疫病由解淀粉欧文氏菌(Burr.)Winslow等人引起,会影响蔷薇科植物,其中包括果园、苗圃和园林种植中的树木和灌木。在2008年和2010年的春夏季节,在土耳其科尼亚省三个不同的景观区域,观察到绣线菊(Spirea prunifolia)出现了枯枝、附着在嫩枝上的坏死叶片以及枯萎的嫩枝。调查期间,植物的发病率约为1%。春季在受感染植物的嫩枝上观察到最初的症状为从微红到褐色的条纹。从七株绣线菊植物嫩枝上的病斑中,在营养蔗糖琼脂(NSA)培养基上分离出九株具有代表性的细菌菌株,并通过生化、生理(2,3)和分子测试(1)鉴定为解淀粉欧文氏菌。细菌为革兰氏阴性、杆状、需氧、发酵型,在米勒和施罗思培养基(2)上呈黄橙色,产左旋糖和3-羟基丁酮呈阳性,在36°C下不生长,明胶水解呈阳性,七叶苷水解、吲哚、脲酶、过氧化氢酶、氧化酶、精氨酸脱氨酶、硝酸盐还原、乳糖产酸和肌醇产酸呈阴性。所有菌株在烟草(Nicotiana tabacum var. White Burley)植株上过敏反应呈阳性。使用物种特异性引物对A/B(1)通过PCR检测以及通过Sherlock微生物鉴定系统软件(TSBA 6 v. 6.00;Microbial ID,纽瓦克,特拉华州)测定的脂肪酸甲酯(FAME)谱,将所有菌株鉴定为解淀粉欧文氏菌,相似性指数范围为79%至99%。通过用含有10 CFU/mL细菌悬浮液的无菌蒸馏水(SDW),使用0.46毫米直径的皮下注射针注射2年生绣线菊品种29号的叶柄和三个正在生长的嫩梢尖来测试致病性。用九种细菌菌株以及两种参考菌株Ea29和NCPPB 2791(土耳其科尼亚塞尔丘克大学植物保护系)分别接种植物。接种后7天,接种植物上出现了与自然感染相关的症状。接种SDW的植物作为阴性对照处理,这些植物上未观察到症状。所有测试重复三次,结果相同。尝试从对照植物以及接种了两种参考菌株和鉴定为解淀粉欧文氏菌的九种细菌的嫩梢和叶片中重新分离细菌。使用上述生化、生理和分子测试,从接种植物中分离出的细菌被鉴定为解淀粉欧文氏菌,但未从对照植物中分离出这种细菌。必须采取植物检疫措施,以避免病原体在土耳其新的景观区域传播到观赏植物上。本报告很重要,因为受感染的绣线菊属植物可能是其他蔷薇科观赏植物的潜在接种源。据我们所知,这是土耳其绣线菊上火疫病发生的首次报告。参考文献:(1)S. Bereswill等人,《应用与环境微生物学》58:3522,1992年。(2)A. L. Jones和K. Geider,《植物病原细菌鉴定实验室指南》,第40 - 55页。美国植物病理学会,明尼苏达州圣保罗,2001年。(3)R. A. Lelliott和D. E. Stead,《植物细菌病害诊断方法(植物病理学方法)》。英国牛津,1987年。