Palou L, Montesinos-Herrero C, Tarazona I, Taberner V
Pathology Laboratory, Postharvest Technology Center, Valencian Institute for Agricultural Research (IVIA), Apartat Oficial, 46113 Montcada, Valencia, Spain.
Plant Dis. 2013 May;97(5):691. doi: 10.1094/PDIS-11-12-1044-PDN.
Commercial production area and yield of sweet persimmon (Diospyros kaki L.) in Spain has doubled in the last 10 years to more than 5,000 ha and 50,000 tons, respectively, mainly because of the high quality and consumer demand for the Valencian autochthonous cultivar 'Rojo Brillante' in European markets. In a recent survey of decay on 'Rojo Brillante' persimmons stored in commercial packinghouses, fruit were found with disease symptoms of firm brown to dark brown round spots scattered on the fruit cheeks. Isolation of the potential causal agent (isolate IVIA QCV-2) was performed by disinfecting the surface of symptomatic fruit with alcohol, aseptically cutting pieces of infected peel tissue, and then plating them onto potato dextrose agar (PDA). The fungus grew fast, covering the entire plate surface (9 mm diameter) after 7 to 10 days of incubation at 25°C with cottony grayish mycelium that darkened with time. Masses of salmon-colored conidia were apparent in the center of some colonies. Conidia were one-celled, hyaline, aseptate, ovoid to oblong with rounded or obtuse ends, and 11.5 to 15.5 × 3.0 to 6.5 μm (n = 50). The identification of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. [synonym: Vermicularia gloeosporioides Penz.; teleomorph: Glomerella cingulata (Stoneman) Spauld. & H. Schrenk] was performed at the Instituto Valenciano de Microbiología (IVAMI, Bétera, Valencia, Spain) by macro and micro morphological observations and confirmed with the amplification and subsequent sequencing of the ribosomal DNA regions 5.8S-ITS2-28S, using the primers ITS3 and ITS4 (4). A representative nucleotide sequence was deposited in GenBank (Accession No. KC113600) and a BLAST search showed 99% identity with the strain C1254.3 of C. gloeosporioides (JX010153) (3). To fulfill Koch's postulates, selected healthy 'Rojo Brillante' persimmons were surface sterilized by dipping them for 2 min in a 0.5% sodium hypochlorite aqueous solution and thoroughly rinsing with fresh water. Mycelial plugs (5-mm diameter) from the edge of 7-day old colonies of isolate IVIA QCV-2 grown on PDA at 25°C were aseptically transferred to skin wounds (one plug per fruit). Wounded but not inoculated fruit were used as controls. Persimmons were placed in three humid chambers that each contained four fruit and incubated at 20°C for up to 21 days. The experiment was repeated twice. While inoculated persimmons developed anthracnose disease in all cases and C. gloeosporioides was consistently reisolated from these fruit, no decay was observed on control fruit. To our knowledge, this is the first report of C. gloeosporioides causing postharvest persimmon fruit rot in Spain. Persimmon anthracnose caused by this pathogen is well known in Asian countries such as China and Korea (1). This disease was also reported in Brazil (2). References: (1) J. H. Lee et al. Plant Pathol. J. 20:247, 2004. (2) M. A. S. Mendes et al. Fungos em Plants no Brasil. Embrapa-SPI/Embrapa-Cenargen, Brasilia, Brazil, 1998. (3) B. S. Weir et al. Stud. Mycol. 73:115, 2012. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press Inc., San Diego, CA, 1990.
在过去10年里,西班牙甜柿(柿属柿树)的商业种植面积和产量分别翻了一番,达到5000多公顷和5万吨以上,这主要是因为欧洲市场对瓦伦西亚本土品种“罗霍·布里兰特”的高品质和消费者需求。在最近一项对商业包装厂储存的“罗霍·布里兰特”柿子腐烂情况的调查中,发现果实出现了坚硬的棕色至深棕色圆形斑点的病害症状,这些斑点散布在果实脸颊上。通过用酒精对有症状果实的表面进行消毒、无菌切割感染的果皮组织块,然后将其接种到马铃薯葡萄糖琼脂(PDA)上,对潜在病原菌(分离株IVIA QCV - 2)进行了分离。该真菌生长迅速,在25°C下培养7至10天后,覆盖了整个平板表面(直径9毫米),形成棉絮状灰白色菌丝体,随着时间推移颜色变深。在一些菌落中心可见大量鲑鱼色分生孢子。分生孢子单细胞、透明、无隔膜、卵形至长方形,两端圆形或钝形,大小为11.5至15.5×3.0至6.5微米(n = 50)。在西班牙瓦伦西亚微生物研究所(IVAMI,贝特拉,瓦伦西亚)通过宏观和微观形态观察对胶孢炭疽菌(Penz.)Penz. & Sacc. [同义词:Vermicularia gloeosporioides Penz.;有性型:Glomerella cingulata (Stoneman) Spauld. & H. Schrenk]进行了鉴定,并使用引物ITS3和ITS4对核糖体DNA区域5.8S - ITS2 - 28S进行扩增和后续测序予以证实(4)。一个代表性核苷酸序列已存入GenBank(登录号KC113600),BLAST搜索显示与胶孢炭疽菌菌株C1254.3(JX010153)有99%的同一性(3)。为了验证科赫法则,选择健康‘罗霍·布里兰特’柿子,将其在0.5%次氯酸钠水溶液中浸泡2分钟进行表面消毒,然后用清水彻底冲洗。将在25°C下PDA上生长7天的分离株IVIA QCV - 2菌落边缘的菌丝块(直径5毫米)无菌转移到果皮伤口处(每个果实一个菌块)。受伤但未接种的果实用作对照。柿子放置在三个湿度箱中,每个湿度箱包含四个果实,在20°C下培养长达21天。该实验重复了两次。虽然接种的柿子在所有情况下都发生了炭疽病,并且始终从这些果实中重新分离出胶孢炭疽菌,但对照果实未观察到腐烂。据我们所知,这是胶孢炭疽菌引起西班牙采后柿果腐烂的首次报道。这种病原菌引起的柿炭疽病在中国和韩国等亚洲国家很常见(1)。巴西也报道过这种病害(2)。参考文献:(1)J. H. Lee等人,《植物病理学报》20:
247,2004。(2)M. A. S. Mendes等人,《巴西植物中的真菌》。巴西农业研究公司 - SPI/巴西农业研究公司 - 巴西农业研究中心,巴西利亚,巴西,1998。(3)B. S. Weir等人,《真菌学研究》73:115,2012。(4)T. J. White等人,见《PCR实验方案:方法与应用指南》第315页。学术出版社,加利福尼亚州圣地亚哥,1990。