Pruvost O, Boyer C, Grygiel P, Boyer K, Verniere C, Gagnevin L, Soro S, N'Guessan C, Kone D
CIRAD-Université de la Réunion, UMR PVBMT, Saint Pierre, La Réunion, F-97410 France.
Université Nangui Abrogoua, Abidjan, Ivory Coast.
Plant Dis. 2014 Dec;98(12):1740. doi: 10.1094/PDIS-07-14-0669-PDN.
Xanthomonas citri pv. mangiferaeindicae causing bacterial canker (or black spot) is a major mango (Mangifera indica L.) pathogen in tropical and subtropical areas (3). The bacterium infects a wide range of mango cultivars, and induces raised, angular, black leaf lesions, sometimes with a yellow chlorotic halo. Fruit symptoms first appear as small water-soaked spots on the lenticels turning into star-shaped, erumpent lesions, which exude an infectious gum, yielding tear-stain patterns. Severe infections cause severe defoliation and/or premature fruit drop. Twig cankers are potential sources of inoculum and weaken branch resistance to winds. Drastic yield losses have been reported at grove scale for susceptible cultivars (3). Mango leaves showing typical angular, black, raised leaf lesions were first observed and collected in April 2014 from trees cv. Kent in five localities of the Korhogo province of Ivory Coast (i.e., the major commercial mango-growing area in this country). Non-pigmented Xanthomonas-like colonies were isolated on KC semi-selective medium (4). Five strains (LL60-1, LL61-1, LL62-1, LL63-1, and LL64-1), one from each locality, were compared by multilocus sequence analysis (MLSA) to the type strain of X. citri and the pathotype strain of several X. citri pathovars, including pvs. anacardii and mangiferaeindicae. This assay targeted the atpD, dnaK, efp, and gyrB genes, as described previously (2). Nucleotide sequences were 100% identical to those of the pathotype strain of X. citri pv. mangiferaeindicae whatever the gene assayed, but differed from any other assayed X. citri pathovar. Leaves of mango cv. Maison Rouge from the youngest vegetative flush were infiltrated (10 inoculation sites/leaf for three replicate leaves on different plants/bacterial strain) as detailed previously (1) with the same five strains. Bacterial suspensions (~1 × 10 cfu/ml) were prepared in 10 mM Tris buffer (pH 7.2) from 16-h-old cultures on YPGA (7 g yeast, 7 g peptone, 7 g glucose, and 18 g agar/liter, pH 7.2). The negative control treatment consisted of three leaves infiltrated with sterile Tris buffer (10 sites/leaf). Plants were incubated in a growth chamber at 30 ± 1°C by day and 26 ± 1°C by night (12-h day/night cycle) at 80 ± 5% RH. All leaves inoculated with the strains from Ivory Coast showed typical symptoms of bacterial canker a week after inoculation. No lesions were recorded from the negative controls. The pathogen was recovered at high population densities (>1 × 10 cfu/lesion) from leaf lesions, typical of a compatible interaction (1) and isolated colonies were identified as the target by atpD sequencing (2). Koch's postulates have therefore been fully verified. This is the first report of the disease in Ivory Coast, a country which has been an internationally significant mango exporter (up to 15,000 tons per year) over the last two decades. A high disease incidence and severity were observed, outlining the need for implementing integrated pest management in mango groves and the production of disease-free nursery stock. This report further expands the distribution of the pathogen in West Africa after its first description from Ghana in 2011 (5) and subsequently in other neighboring countries. References: (1) N. Ah-You et al. Phytopathology 97:1568, 2007. (2) L. Bui Thi Ngoc et al. Int. J. Syst. Evol. Microbiol. 60:515, 2010. (3) L. Gagnevin and O. Pruvost. Plant Dis. 85:928, 2001. (4) O. Pruvost et al. J. Appl. Microbiol. 99:803, 2005. (5) O. Pruvost et al. Plant Dis. 95:774, 2011.
引起细菌性溃疡病(或黑斑病)的芒果黄单胞菌芒果致病变种是热带和亚热带地区芒果(杧果)的一种主要病原菌(3)。该细菌可感染多种芒果品种,会导致叶片出现凸起的、角状的黑色病斑,有时病斑周围还伴有黄色褪绿晕圈。果实症状最初表现为皮孔上出现小的水渍状斑点,随后变为星状、突出的病斑,病斑会渗出具有传染性的树胶,形成泪痕状图案。严重感染会导致严重落叶和/或果实过早脱落。嫩枝溃疡病斑是潜在的接种源,会削弱枝条的抗风能力。据报道,对于易感品种,果园规模的产量会大幅损失(3)。2014年4月,首次在科特迪瓦省科霍戈市的五个地区(即该国主要的商业芒果种植区),从肯特品种的树上观察并采集到了表现出典型角状、黑色、凸起叶片病斑的芒果叶片。在KC半选择性培养基上分离出了无色的类黄单胞菌菌落(4)。通过多位点序列分析(MLSA),将从每个地区分离得到的5个菌株(LL60 - 1、LL61 - 1、LL62 - 1、LL63 - 1和LL64 - 1)与柑橘黄单胞菌的模式菌株以及几种柑橘黄单胞菌致病型菌株(包括杧果致病变种和腰果致病变种)进行了比较。该分析针对atpD、dnaK、efp和gyrB基因,方法如前所述(2)。无论检测哪个基因,核苷酸序列都与芒果黄单胞菌致病型菌株的序列100%相同,但与其他检测的柑橘黄单胞菌致病型变种不同。按照之前详细描述的方法(1),用同样的5个菌株对来自最幼嫩营养枝梢的芒果品种“红房子”的叶片进行浸润接种(每片叶10个接种位点,不同植株/细菌菌株的3片重复叶)。从在YPGA(7 g酵母、7 g蛋白胨、7 g葡萄糖和18 g琼脂/升,pH 7.2)上培养16小时的培养物中,在10 mM Tris缓冲液(pH 7.2)中制备细菌悬浮液(约1×10 cfu/ml)。阴性对照处理包括用无菌Tris缓冲液浸润3片叶(每片叶10个位点)。将植株置于生长室中,白天温度为30±1°C,夜间温度为26±1°C(12小时昼夜循环),相对湿度为80±5%。接种来自科特迪瓦的菌株的所有叶片在接种一周后都表现出典型的细菌性溃疡病症状。阴性对照未记录到病斑。从叶片病斑中以高菌量密度(>1×10 cfu/病斑)分离到病原菌,这是典型的亲和性互作(1),通过atpD测序鉴定分离菌落为目标病原菌(2)。因此,柯赫氏法则得到了充分验证。这是该病在科特迪瓦的首次报道,在过去二十年里,该国一直是具有国际影响力的芒果出口国(每年高达15000吨)。观察到该病的高发病率和严重程度,凸显了在芒果园中实施综合病虫害管理以及生产无病苗木的必要性。本报告进一步扩展了该病原菌在西非的分布范围,此前它于2011年在加纳首次被描述(5),随后在其他邻国也有发现。参考文献:(1)N. Ah - You等人,《植物病理学》97:1568,2007年。(2)L. Bui Thi Ngoc等人,《国际系统与进化微生物学杂志》60:515,2010年。(3)L. Gagnevin和O. Pruvost,《植物病害》85:928,2001年。(4)O. Pruvost等人,《应用微生物学杂志》99:803,2005年。(5)O. Pruvost等人,《植物病害》95:774,2011年。