Girard J-C, Noëll J, Larbre F, Roumagnac P, Rott P
CIRAD, UMR BGPI, F-34398 Montpellier, France.
SUCAF GABON, BP610, Franceville, Gabon.
Plant Dis. 2014 May;98(5):684. doi: 10.1094/PDIS-09-13-0914-PDN.
During a disease inspection at the sugarcane estate SUCAF near Franceville, Gabon, in March 2011, 1- to 3-mm wide and several dm long dark red stripes were observed on sugarcane (Saccharum spp.) leaves of many plants of cultivar R570. These plants were 5.5 months old in the first ratoon crop. Additionally, spindle leaves of several stalks were rotted and could be easily pulled out of the top of the stalk. Longitudinal sections of diseased stalks showed reddish-brown discoloration of the upper stem and the rotted spindle had an unpleasant odor. Circular, convex, smooth, yellow-cream pigmented bacterial colonies with 2 to 3 mm diameter were isolated after 3 days at 28°C from young leaf lesions on YPGA (yeast extract 7 g/L, peptone 7 g/L, glucose 7 g/L, agar 15 g/L, pH 6.8 to 7.0). The 16S-23S internal transcribed spacer (ITS) of two representative colonies was PCR amplified, and the nucleotide sequences were shown to be 99% identical to the 16S-23S ITS sequence from the genome of Acidovorax avenae subsp. avenae strain ATCC 19860 (GenBank: CP002521.1). One of these A. avenae subsp. avenae isolates from Gabon was inoculated into greenhouse grown plants of sugarcane cultivar R570. Plants were inoculated by injection into the sheath of spindle leaves above the meristem with the bacterial strain (12 plants) or with a water control (six plants). In this method, the bacteria (10 CFU/ml) were injected using a syringe through the leaf sheath until filling the leaf whorl. Three weeks post-inoculation, one to several cm long red-brown stripes were observed on leaves of 11 of 12 inoculated plants. Seven weeks post-inoculation, all plants exhibited symptoms, from red, brown, or black stripes to leaf necrosis, rotting, and death of the spindle leaves (six plants). All six control plants were symptomless. In a second experiment, 6 of 12 plants showed symptoms 3 weeks post inoculation, and the pathogen was successfully re-isolated from all six symptomatic plants with YPGA medium. The 16S-23S ITS of three single colonies obtained each from different symptomatic plants were PCR amplified and the nucleotide sequences were again found 99% identical to the 16S-23S ITS sequence from the genome of A. avenae subsp. avenae ATCC 19860. To our knowledge, this is the first report of A. avenae subsp. avenae, the causal agent of sugarcane red stripe (also reported as top rot), in Gabon. It is also the first description of the occurrence of the top rot form of the disease in R570, a cultivar that is grown in several locations of Africa, the Mascarene Islands, and the French West Indies. A large-scale survey needs to be undertaken to determine the distribution of red stripe in Gabon, a disease for which several outbreaks have been reported recently worldwide (1,2). References: (1) M. P. Grisham and R. M. Johnson. Phytopathology 101:564, 2011. (2) S. Zia-ul-Hussnain et al. Afr. J. Biotechnol. 10:7191, 2011.
2011年3月,在加蓬弗朗斯维尔附近的SUCAF甘蔗种植园进行病害检查时,在许多R570品种甘蔗(甘蔗属)植株的叶片上观察到宽1至3毫米、长数分米的深红色条纹。这些植株处于头茬宿根作物阶段,生长了5.5个月。此外,几根茎的纺锤叶腐烂,很容易从茎顶拔出。患病茎的纵切面显示上部茎呈红棕色变色,腐烂的纺锤叶有难闻气味。在YPGA(酵母提取物7 g/L、蛋白胨7 g/L、葡萄糖7 g/L、琼脂15 g/L,pH 6.8至7.0)培养基上,于28°C培养3天后,从幼叶病斑中分离出直径2至3毫米的圆形、凸起、光滑、黄色奶油色的细菌菌落。对两个代表性菌落的16S - 23S内部转录间隔区(ITS)进行PCR扩增,结果显示其核苷酸序列与燕麦嗜酸菌燕麦亚种ATCC 19860基因组的16S - 23S ITS序列有99%的同一性(GenBank:CP002521.1)。从加蓬分离得到的其中一株燕麦嗜酸菌燕麦亚种被接种到温室种植的R570品种甘蔗植株上。通过将细菌菌株(12株植株)或水对照(6株植株)注射到分生组织上方纺锤叶的叶鞘中来接种植株。采用这种方法,使用注射器通过叶鞘注射细菌(10 CFU/ml),直至充满叶环。接种后三周,12株接种植株中有11株的叶片上出现了1至数厘米长的红棕色条纹。接种后七周,所有植株都出现了症状,从红色、棕色或黑色条纹到叶片坏死、腐烂以及纺锤叶死亡(6株植株)。所有6株对照植株均无症状。在第二个实验中,12株植株中有6株在接种后三周出现症状,并且从所有6株有症状的植株上成功地用YPGA培养基重新分离出了病原菌。从不同有症状植株上分别获得的三个单菌落的16S - 23S ITS进行PCR扩增,再次发现其核苷酸序列与燕麦嗜酸菌燕麦亚种ATCC 19860基因组的16S - 23S ITS序列有99%的同一性。据我们所知,这是燕麦嗜酸菌燕麦亚种在加蓬的首次报道,燕麦嗜酸菌燕麦亚种是甘蔗红条纹病(也被报道为顶腐病)的病原菌。这也是对R570品种中顶腐病形式发生情况的首次描述,R570品种在非洲、马斯克林群岛和法属西印度群岛的多个地方种植。需要进行大规模调查以确定加蓬红条纹病的分布情况,最近在全球范围内已报道了该病害的几次爆发(参考文献1,2)。参考文献:(1) M. P. Grisham和R. M. Johnson。《植物病理学》101:564,2011年。(2) S. Zia - ul - Hussnain等人。《非洲生物技术杂志》10:7191,2011年。