Ferreira A P S, Pinho D B, Machado A R, Pereira O L
Departamento de Fitopatologia, Universidade Federal de Viçosa, Viçosa, Minas Gerais, 36570-900, Brazil.
Plant Dis. 2014 Sep;98(9):1277. doi: 10.1094/PDIS-03-14-0288-PDN.
Pineapple (Ananas comosus L. Merril.) is the main plant of the Bromeliaceae, cultivated economically for the fruits' appealing flavor and a refreshing sugar-acid balance. In 2013, fruits with no initially visible symptoms began to show a postharvest rot after 3 days in a market in the municipality of Viçosa, Minas Gerais, Brazil. The rot can rarely be detected from the outside of the fruit, but a longitudinal section allows observation of extension of the affected area toward the center of the fruit. The symptoms initially appear as a dark brown to black rot on surface of the fruits, which gradually enlarges in size, leading to increased rot and disposal of infected fruits. Until now, this disease occurred sporadically and caused small losses. A fungus was isolated from rot observed in fruits from cultivar Pérola and a single-spore culture was deposited in the culture collection of the Universidade Federal de Viçosa (Accession No. COAD 1588). After 7 days of incubation at 25°C, the strain displayed radial growth and gray-white to black colonies. Microscopic observations revealed brown to light brown conidiophores present singly or in groups. The septate, simple or rarely branched conidiophores are straight or curved, up to 245 μm long and 5 μm wide, and some have a geniculate growth pattern near the apex. The conidia are ellipsoidal or barrel-shaped and 22 to 25 μm long and 10 to 12.5 μm wide. The median septum appears as a black band and the cells at each end of the conidia are pale, whereas the intermediate cells are brown or dark brown. Based on morphological characteristics, the fungus was identified as Curvularia eragrostidis (4). To confirm this identification, DNA was extracted and sequences of the internal transcribed spacer (ITS), 28S and 18S rDNA regions were obtained and deposited in GenBank (Accession Nos. KJ541818 to KJ541820). The sequence of the ITS region exhibited 99% identity over 530 bp with other C. eragrostidis sequence in GenBank (JN943449) and Bayesian inference analysis placed our isolate in the same clade with others C. eragrostidis (study S15670 deposited in TreeBASE). Koch's postulates were conducted by inoculating six fruits of pineapple previously disinfected with 2% sodium hypochlorite and washed in sterile distilled water. For inoculation, the isolate was grown in potato dextrose agar (PDA) for 15 days at 25°C. Six millimeter diameter disks were removed from the surface of fruits with a sterile cork borer and replaced with PDA disks containing mycelia from the margins of the culture. An agar plug was deposited in three control fruits and all fruits were maintained at 25°C in plastic trays. Inoculated fruits showed symptoms 7 days after inoculation that were similar to those initially observed in the infected fruits, while control fruits showed no symptoms. C. eragrostidis is a cosmopolitan pathogen that infects hosts from several botanical families (2,4). In Brazil, this fungus causes leaf spot on A. comosus (3) and also infects Allium sativum, Dioscorea alata, D. cayenensis, Oryza sativa, Sorghum bicolor, Vigna unguiculata, and Zea mays (1). To our knowledge, this is the first report of C. eragrostidis causing postharvest rot disease in pineapple in Brazil. Because invasion of the fungus can occur through minute fractures, fruits should be carefully handled to avoid mechanical damage. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases , 18 February 2014. (2) D. S. Manamgoda et al. Fungal Divers. 51:3, 2011. (3) J. J. Ponte et al. Fitopatologia 10:21, 1975. (4) A. Sivanesan. Mycological Papers 158:113, 1987.
菠萝(Ananas comosus L. Merril.)是凤梨科的主要植物,因其果实诱人的风味和清爽的糖酸平衡而被广泛种植用于经济目的。2013年,在巴西米纳斯吉拉斯州维索萨市的一个市场上,最初没有明显症状的菠萝在收获后3天开始出现采后腐烂。这种腐烂在果实外部很难检测到,但纵切可以观察到受影响区域向果实中心扩展。症状最初表现为果实表面深褐色至黑色的腐烂,随后逐渐扩大,导致腐烂加剧,受感染果实被丢弃。到目前为止,这种病害偶尔发生,造成的损失较小。从佩罗拉品种果实上观察到的腐烂部位分离出一种真菌,并将单孢子培养物保存在维索萨联邦大学的培养物保藏中心(保藏编号COAD 1588)。在25°C下培养7天后,该菌株呈现出径向生长,菌落为灰白色至黑色。显微镜观察显示,褐色至浅褐色的分生孢子梗单个或成簇出现。有隔膜、简单或很少分枝的分生孢子梗直或弯曲,长达245μm,宽5μm,有些在顶端附近呈膝状生长模式。分生孢子呈椭圆形或桶形,长22至25μm,宽10至12.5μm。中间隔膜呈现为黑色带,分生孢子两端的细胞颜色较浅,而中间细胞为褐色或深褐色。根据形态特征,该真菌被鉴定为画眉草弯孢霉(4)。为了证实这一鉴定,提取了DNA,并获得了内部转录间隔区(ITS)、28S和18S rDNA区域的序列,并保存在GenBank中(保藏编号KJ541818至KJ541820)。ITS区域的序列在530 bp上与GenBank中其他画眉草弯孢霉序列(JN943449)的同一性为99%,贝叶斯推断分析将我们的分离株与其他画眉草弯孢霉置于同一分支中(保存在TreeBASE中的研究S15670)。通过接种六个预先用2%次氯酸钠消毒并在无菌蒸馏水中冲洗过的菠萝果实来进行柯赫氏法则验证。为了接种,将分离株在马铃薯葡萄糖琼脂(PDA)上于25°C培养15天。用无菌打孔器从果实表面取下直径6毫米的圆盘,并用含有来自培养物边缘菌丝体的PDA圆盘替换。在三个对照果实中放置一个琼脂块,所有果实均保存在25°C的塑料托盘中。接种后的果实接种7天后出现了与最初在受感染果实中观察到的症状相似的症状,而对照果实没有出现症状。画眉草弯孢霉是一种世界性病原菌,可感染多个植物科的寄主(2,4)。在巴西,这种真菌会导致菠萝叶斑病(3),还会感染大蒜、薯蓣、山药、水稻、高粱、豇豆和玉米(1)。据我们所知,这是巴西首次报道画眉草弯孢霉引起菠萝采后腐烂病。由于真菌可通过微小的裂缝侵入,因此应小心处理果实以避免机械损伤。参考文献:(1)D. F. Farr和A. Y. Rossman。真菌数据库。美国农业部农业研究局系统真菌学和微生物学实验室。检索自http://nt.ars-grin.gov/fungaldatabases,2014年2月18日。(2)D. S. Manamgoda等人。真菌多样性。51:3,2011年。(3)J. J. Ponte等人。植物病理学10:21,1975年。(4)A. Sivanesan。真菌学论文158:113,1987年。