Yasuhara-Bell J, de Silva A S, Alvarez A M, Shimabuku R, Ko M
Department of Plant and Environmental Protection Sciences, University of Hawai'i at Mānoa, Honolulu, O'ahu 96822.
College of Tropical Agriculture and Human Resources, Cooperative Extension Service, University of Hawai'i at Mānoa, Kahului, Maui 96732.
Plant Dis. 2013 Sep;97(9):1244. doi: 10.1094/PDIS-11-12-1071-PDN.
Bacterial black spot of mango (Mangifera indica) caused by Xanthomonas citri pv. mangiferaeindicae (Xcm), is an economically important disease in tropical and subtropical areas (3). Xcm can infect a wide range of mango cultivars and induces raised, angular, black leaf lesions, sometimes with a chlorotic halo. Fruit symptoms appear as small, water-soaked spots on the lenticels that become star-shaped, erumpent, and exude an infectious gum (3). The bacterium can also cause latent infections (2). Immature mango fruit with black spots on the epidermis were collected in August 2012 from mango trees of the cvs. Raposa and Pirie at a residence in Pukalani, Hawai'i, on the island of Maui. Similar symptoms were seen on a tree of the mango cv. Common (also known as 'Spanish' or 'Lahaina') at a nearby golf course. Mango fruit with black lesions, and leaves showing black lesions with yellow halos, were collected in August 2012 from mango trees of the cv. Haden at a residence in Kaimuki, Hawai'i, on the island of O'ahu. Xanthomonas-like bacterial colonies were isolated on TZC agar. Suspect colonies were non-pigmented on YDC agar. A fruit strain of the bacterium from Maui (A6081A) and a strain from each of a fruit (A6081B) and a leaf (A6082) from O'ahu were each gram-negative, oxidative, positive for both starch and esculin hydrolysis, and negative for nitrate reduction, resulting in presumptive identification as a Xanthomonas sp. The three strains were further characterized by Microlog (Biolog, Inc. Hayward, CA), which showed the closest match with X. campestris. In addition, 16S rDNA PCR assays showed the closest match (99% similarity) with X. citri strains, and RIF marker analysis of dnaA (4) grouped the three strains with Xcm strain LMG 941 (Accession No. CAHO01000002.1). Hypersensitivity responses typical of xanthomonads were observed when these strains were infiltrated into tobacco leaves, whereas no response was observed using sterile water. Leaves of 3-week-old mango seedlings were infiltrated using 10 μl (~10 CFU/ml) of each strain suspended in sterilized water (six to eight inoculations per leaf, four leaves per plant, and three replicate plants per strain). The negative control treatments consisted of inoculation with sterile water, as well as an incompatible pathogen, X. hortorum pv. vitians (A6076), isolated from lettuce. Typical symptoms of bacterial black spot were observed for all strains assayed approximately 2 weeks after inoculation. No lesions were observed on the negative control plants. Koch's postulates were satisfied following reisolation and identification of the Xanthomonas strains from the infected plant tissues, using the biochemical and PCR methods described above. Results for strains from the two islands confirmed published descriptions of the pathogen, indicating that the pathogen causing symptoms on these mango trees is Xcm (1). Cultures and infected plant samples were sent to USDA APHIS and CPHST NPGLB facilities where this identification was confirmed. To our knowledge, this is the first report of bacterial black spot of mango in Hawai'i or anywhere in the United States. It is unknown whether this disease is a new occurrence or has not been reported previously. The origin of the primary inoculum is unknown. References: (1) B. Manicom and F. Wallis. Int. J. Syst. Bacteriol. 34:77, 1984. (2) O. Pruvost et al. Microbial Ecol. 58:928. (3) O. Pruvost et al. Plant Dis. 95:774, 2011. (4) K. Schneider et al. PLoS 6:e18496, 2011.
由柑桔黄单胞菌芒果致病变种(Xcm)引起的芒果细菌性黑斑病,是热带和亚热带地区一种具有经济重要性的病害(3)。Xcm能感染多种芒果品种,可诱发叶片出现凸起的、角状的黑色病斑,有时病斑周围还有黄化晕圈。果实症状表现为皮孔上出现小的水渍状斑点,这些斑点会变成星形、突出,并渗出具传染性的树胶(3)。该细菌还能引发潜伏感染(2)。2012年8月,从毛伊岛普卡拉尼一处住宅的拉波萨和皮里品种芒果树上采集了表皮有黑斑的未成熟芒果果实。在附近一个高尔夫球场的一棵普通芒果品种(也称为“西班牙”或“拉海纳”)树上也发现了类似症状。2012年8月,从瓦胡岛凯穆基一处住宅的哈登品种芒果树上采集了有黑色病斑的芒果果实以及带有黄色晕圈黑色病斑的叶片。在TZC琼脂上分离出了类似黄单胞菌的菌落。疑似菌落在YDC琼脂上不产生色素。从毛伊岛分离得到的一个果实菌株(A6081A)以及从瓦胡岛分离得到的一个果实菌株(A6081B)和一个叶片菌株(A6082)均为革兰氏阴性、氧化型,淀粉和七叶苷水解试验呈阳性,硝酸盐还原试验呈阴性,初步鉴定为黄单胞菌属细菌。通过微生物鉴定系统(Microlog,Biolog公司,加利福尼亚州海沃德市)对这三个菌株进行进一步鉴定,结果显示与野油菜黄单胞菌最为匹配。此外,16S rDNA PCR检测表明与柑桔黄单胞菌菌株相似度最高(99%),对dnaA基因的利福平标记分析(4)将这三个菌株与Xcm菌株LMG 941(登录号CAHO01000002.1)归为一类。当将这些菌株注射到烟草叶片中时,观察到了典型的黄单胞菌过敏反应,而用无菌水注射则未观察到反应。将三周龄芒果幼苗的叶片用10 μl(约10 CFU/ml)悬浮于无菌水中的每个菌株进行注射(每片叶接种六至八次,每株植物四片叶,每个菌株三个重复植株)。阴性对照处理包括用无菌水接种以及接种从生菜中分离得到的非亲和病原菌——莴苣黄单胞菌(A6076)。接种后约两周,对所有检测菌株均观察到了细菌性黑斑病的典型症状。阴性对照植株未出现病斑。使用上述生化和PCR方法,从感染植物组织中重新分离并鉴定出黄单胞菌菌株后,科赫法则得到了满足。来自两个岛屿的菌株检测结果证实了已发表的关于该病原菌的描述,表明在这些芒果树上引起症状的病原菌是Xcm(1)。培养物和感染植物样本被送往美国农业部动植物卫生检验局和植物卫生检验局国家植物种质资源库实验室,在那里该鉴定结果得到了确认。据我们所知,这是夏威夷或美国任何地方首次报道芒果细菌性黑斑病。尚不清楚这种病害是新出现的还是以前未被报道过。初始接种体的来源不明。参考文献:(1)B. Manicom和F. Wallis。《国际系统细菌学杂志》34:77,1984年。(2)O. Pruvost等人。《微生物生态学》58:928。(3)O. Pruvost等人。《植物病害》95:774,2011年。(4)K. Schneider等人。《公共科学图书馆·综合》6:e18496,2011年。