Jouen E, Robène-Soustrade I, Gagnevin L, Pruvost O, Benimadhu S
CIRAD-Université de la Réunion, UMR PVBMT, Saint Pierre, La Réunion, F-97410 France.
AREU, Plant Pathology Division, Réduit, Mauritius.
Plant Dis. 2008 Jun;92(6):980. doi: 10.1094/PDIS-92-6-0980A.
In November of 2006, necrotic leaf lesions with water-soaked margins were observed on Syngonium podophyllum in Floréal, Forest Side, and Réduit, Mauritius. Although not an economically important crop, the disease was of concern because syngonium is a host for Xanthomonas axonopodis pv. dieffenbachiae and the anthurium industry is of major economic importance in Mauritius. X. campestris pv. syngonii, described as the causal agent of bacterial leaf blight of syngonium (2), is genetically closely related to group 9.4 X. axonopodis pv. dieffenbachiae strains (3). In contrast to X. axonopodis pv. dieffenbachiae, X. campestris pv. syngonii strains are highly virulent on syngonium but are not pathogenic on anthurium or other Araceae, but both react similarly to the Xcd108 monoclonal antibody (Mab) (Agdia Inc., Elkhart, IN) and to a nested PCR assay designed for X. axonopodis pv. dieffenbachiae (4). X. axonopodis pv. dieffenbachiae and X. campestris pv. syngonii strains can be distinguished on the basis of restriction analysis of the amplicon of this PCR assay. Four pure cultures isolated from S. podophyllum were gram negative, yellow pigmented, and produced mucoid colonies on yeast peptone glucose agar (YPGA). One positive control strain of X. campestris pv. syngonii (LMG 9055 from the United States) and X. axonopodis pv. dieffenbachiae (LMG 695 from Brazil) were also used for all tests. All strains reacted positively with the Xcd108 MAb using indirect ELISA. DNA from all strains was amplified by the nested PCR assay, and the HincII restriction pattern of the amplicons identified strains from Mauritius as X. campestris pv. syngonii. Pathogenicity tests were performed on 8-month-old plants of Anthurium andreanum cv. Florida, Dieffenbachia maculata cv. Tropic Marianne, and S. podophyllum cv. Robusta by infiltrating suspensions containing ~1 × 10 CFU ml¯ of each strain prepared from YPGA plates. Each strain was inoculated onto three young leaves (four inoculation sites per leaf) on two plants. Negative control plants received sterile Tris buffer solution (10 mM, pH 7.2). Plants were maintained in a growth chamber with day and night temperatures at 30 ± 1°C and 26 ± 1°C, respectively, 95 ± 5% relative humidity, 30 μmol·m¯·s¯ light intensity, and a photoperiod of 12 h (4). All strains caused typical water-soaked lesions 14 days after inoculation (dai) on syngonium. Lesions turned necrotic with chlorotic margins 27 to 34 dai. Typical bacterial blight lesions were observed on anthurium leaves inoculated with X. axonopodis pv. dieffenbachiae strain LMG 695, but no symptoms were observed 60 dai when strains from Mauritius and LMG 9055 were used. Amplified fragment length polymorphism analysis of four strains from Mauritius and additional reference, X. axonopodis pv. dieffenbachiae and X. campestris pv. syngonii strains, using SacI/MspI and four primer pairs (unlabeled MspI+1 [A, C, T, or G] primers and 5'-labeled-SacI+C primer for the selective amplification step) (1), showed that the strains from Mauritius could be distinguished from X. axonopodis pv. dieffenbachiae but were identical to X. campestris pv. syngonii strains from the United States and Réunion Island. References: (1) N. Ah-You et al. Phytopathology 97:1568, 2007. (2) R. S. Dickey and C. H. Zumoff. Phytopathology 77:1257, 1987. (3) J. L. W. Rademaker et al. Phytopathology 95:1098, 2005. (4) I. Robene-Soustrade et al. Appl. Environ. Microbiol. 72:1072, 2006.
2006年11月,在毛里求斯弗洛雷亚尔、森林边和雷迪特地区的箭叶芋上观察到边缘有水渍状的坏死叶斑。虽然箭叶芋不是经济作物,但这种病害令人担忧,因为箭叶芋是野油菜黄单胞菌致病变种的寄主,而红掌产业在毛里求斯具有重要经济意义。野油菜黄单胞菌箭叶芋致病变种被描述为箭叶芋细菌性叶斑病的病原菌(2),在基因上与9.4组野油菜黄单胞菌致病变种菌株密切相关(3)。与野油菜黄单胞菌致病变种不同,野油菜黄单胞菌箭叶芋致病变种菌株对箭叶芋具有高度致病性,但对红掌或其他天南星科植物无致病性,不过二者对Xcd108单克隆抗体(Mab)(Agdia公司,美国印第安纳州埃尔克哈特)和针对野油菜黄单胞菌致病变种设计的巢式PCR检测反应相似(4)。野油菜黄单胞菌致病变种和野油菜黄单胞菌箭叶芋致病变种菌株可根据该PCR检测扩增产物的限制性分析加以区分。从箭叶芋分离得到的4个纯培养物革兰氏阴性,产黄色色素,在酵母蛋白胨葡萄糖琼脂(YPGA)上形成黏液状菌落。野油菜黄单胞菌箭叶芋致病变种的1个阳性对照菌株(来自美国的LMG 9055)和野油菜黄单胞菌致病变种(来自巴西的LMG 695)也用于所有检测。所有菌株通过间接ELISA与Xcd108 MAb反应呈阳性。所有菌株的DNA通过巢式PCR检测进行扩增,扩增产物的HincII限制性图谱将来自毛里求斯的菌株鉴定为野油菜黄单胞菌箭叶芋致病变种。通过将从YPGA平板制备的每种菌株的约1×10 CFU/ml悬浮液注入8月龄的红掌品种佛罗里达、花叶万年青品种热带玛丽安和箭叶芋品种健壮植株进行致病性检测。每个菌株接种到两株植物的三片幼叶上(每片叶四个接种部位)。阴性对照植株接种无菌Tris缓冲液(10 mM,pH 7.2)。植株置于生长室中,白天和夜间温度分别为30±1°C和26±1°C,相对湿度95±5%,光照强度30 μmol·m¯·s¯,光周期12小时(4)。所有菌株在接种后14天(dai)在箭叶芋上引起典型的水渍状病斑。病斑在接种后27至34天变为坏死,边缘黄化。在用野油菜黄单胞菌致病变种菌株LMG 695接种的红掌叶片上观察到典型的细菌性叶斑病病斑,但使用来自毛里求斯的菌株和LMG 9055接种时,在接种后60天未观察到症状。使用SacI/MspI和四对引物(未标记的MspI + 某一种碱基 [A、C、T或G] 引物以及用于选择性扩增步骤的5'-标记-SacI + C引物)(1)对来自毛里求斯的4个菌株以及其他参考菌株、野油菜黄单胞菌致病变种和野油菜黄单胞菌箭叶芋致病变种菌株进行扩增片段长度多态性分析,结果表明来自毛里求斯的菌株可与野油菜黄单胞菌致病变种区分开,但与来自美国和留尼汪岛的野油菜黄单胞菌箭叶芋致病变种菌株相同。参考文献:(1)N. Ah-You等人,《植物病理学》97:1568,2007年。(2)R. S. Dickey和C. H. Zumoff,《植物病理学》77:1257,1987年。(3)J. L. W. Rademaker等人,《植物病理学》9:1098,2005年。(4)I. Robene-Soustrade等人,《应用与环境微生物学》72:1072,2006年。