Vakalounakis D J, Doulis A G, Lamprou K K
Laboratory of Phytopathological Mycology, Plant Protection Institute, Hellenic Agricultural Organization 'Demeter' (NAGREF), P.O. Box 2228, GR-710 03 Heraklio, Greece.
Laboratory of Plant Biotechnology, Institute of Viticulture, Floriculture and Vegetable Crops, Hellenic Agricultural Organization 'Demeter' (NAGREF), P.O. Box 2228, GR-71003, Heraklio, Greece.
Plant Dis. 2014 Mar;98(3):425. doi: 10.1094/PDIS-07-13-0776-PDN.
In February 2013, severe symptoms of the powdery scab canker type were observed during harvesting at the Mallia area of Heraklio, Crete, Greece, on approximately 20% of the tubers of two commercial smooth skinned potato (Solanum tuberosum L.) field crops (cv. Spunta), of which the seeds had been imported from the Netherlands. Initially, small (up to 2 mm diameter), sunken, purple-brown lesions appeared before harvest on the tubers, followed by pimple-like swellings, which increased in size. As lesions matured, they broke through the tuber skin and developed into shallow depressions. Lesions often expanded in depth and width, forming deep pits and open cankers, eventually destroying considerable amounts of tuber tissue. Light microscopic preparations showed the presence of spongy spore balls with a honeycomb-like spore wall structure. Spore balls were golden brown to brown, oval, circular, elongate, pyriform or irregular in shape, and 54.2 ± 0.93 (36.3 to 79.9) × 47.8 ± 0.82 (31.5 to 72.6) μm. These were composed of aggregates of hundreds of spores that were circular to polygonal in shape, and 4.5 ± 0.06 (3.5 to 5.6) μm in diameter. The morphological characteristics closely resembled those reported for the powdery scab pathogen Spongospora subterranea (Wallr.) Lagerheim f. sp. subterranea Tomlinson. Pathogenicity was confirmed by a lab-based bioassay (3), where spore balls scraped from tuber scab lesions were used to inoculate 20 tomato (cv. Earlypak) bait seedlings, while 20 other seedlings were employed as non-inoculated controls. Zoosporangia of S. subterranea were observed microscopically in root hairs and epidermal cells of all inoculated seedlings but not of any control plants 2 weeks after inoculation. The identity of the pathogen was also confirmed in the original potato samples employing Sss AgriStrip (BIOREBA AG, Reinach, Switzerland), an immunological assay specific for S. subterranea resting spores. In addition, DNA was extracted from infected tubers and the region corresponding to the internal transcribed spacers (ITS) and the ribosomal DNA (rDNA) gene regions was amplified (1). PCR products were purified, direct sequenced, and deposited in GenBank (Accession No. KF208654). In a BLASTn search, the 18S (partial)-ITS1-5.8S-ITS2 (partial) sequence stretch showed 95% similarity to a 509-bp sequence (AF102820.1) of the corresponding S. subterranea gene in GenBank (query coverage 98% and E value 0.0). Based on morphological characteristics, pathogenicity test, immunological assay, and molecular data, it was concluded that the pathogen on potato in Crete is S. subterranea f. sp. subterranea. Powdery scab of potatoes by S. subterranea had been reported earlier in some areas of Greece (Argolida, Attica, Kefallinia, Messenia, and Salonika) (2,4), but without any data confirming the identity of the pathogen. To our knowledge, this is the first record in the island of Crete, and the first confirmed report of this pathogen in Greece. Because S. subterranea f. sp. subterranea is a destructive pathogen of potatoes, its presence in Crete could be of significant concern for commercial growers. Consequently seed import regulations should be strictly enforced. References: (1) K. S. Bell et al. Eur. J. Plant Path. 105:905, 1999. (2) C. D. Holevas et al. Ann. Inst. Phytopath. Benaki 19:52, 2000. (3) U. Merz. Bull. OEPP 19:585, 1989. (4) M. E. Pantidou. Fungus-host index for Greece. Benaki Phytopath. Inst., Athens, 1973.
2013年2月,在希腊克里特岛伊拉克利翁的马利亚地区收获期间,观察到约20%的两个商业种植的光滑皮马铃薯(Solanum tuberosum L.)大田作物(品种为Spunta)块茎出现粉状痂溃疡型严重症状,这些作物的种子从荷兰进口。收获前,块茎上最初出现小的(直径达2毫米)、凹陷的紫褐色病斑,随后出现丘疹样肿胀,且肿胀面积增大。随着病斑成熟,它们穿透块茎表皮,发展成浅凹陷。病斑常常在深度和宽度上扩展,形成深坑和开放性溃疡,最终破坏大量块茎组织。光学显微镜制片显示存在具有蜂窝状孢子壁结构的海绵状孢子球。孢子球呈金褐色至褐色,椭圆形、圆形、细长形、梨形或不规则形,大小为54.2±0.93(36.3至79.9)×47.8±0.82(31.5至72.6)微米。它们由数百个圆形至多边形的孢子聚集而成,孢子直径为4.5±0.06(3.5至5.6)微米。这些形态特征与粉状痂病原菌Spongospora subterranea (Wallr.) Lagerheim f. sp. subterranea Tomlinson报道的特征极为相似。通过基于实验室的生物测定(3)证实了致病性,从块茎痂病斑刮下的孢子球用于接种20株番茄(品种为Earlypak)诱饵幼苗,同时另外20株幼苗作为未接种对照。接种2周后,在所有接种幼苗的根毛和表皮细胞中显微镜观察到S. subterranea的游动孢子囊,但对照植株中未观察到。还采用Sss AgriStrip(瑞士雷纳赫的BIOREBA AG公司)在原始马铃薯样品中证实了病原菌的身份,这是一种针对S. subterranea休眠孢子的免疫测定。此外,从受感染块茎中提取DNA,并扩增对应于内转录间隔区(ITS)和核糖体DNA(rDNA)基因区域(1)。PCR产物经纯化、直接测序,并保存在GenBank(登录号KF208654)。在BLASTn搜索中,18S(部分)-ITS1-5.8S-ITS2(部分)序列片段与GenBank中相应的S. subterranea基因的509碱基对序列(AF102820.1)显示95%的相似性(查询覆盖率98%,E值0.0)。基于形态特征、致病性测试、免疫测定和分子数据,得出结论:克里特岛马铃薯上的病原菌为S. subterranea f. sp. subterranea。希腊的一些地区(阿哥利达、阿提卡、凯法利尼亚、梅西尼亚和萨洛尼卡)(2,4)此前曾报道过S. subterranea引起的马铃薯粉状痂病,但没有任何数据证实病原菌的身份。据我们所知,这是克里特岛的首次记录,也是希腊该病原菌的首次确诊报告。由于S. subterranea f. sp. subterranea是马铃薯的一种毁灭性病原菌,其在克里特岛的出现可能引起商业种植者的极大关注。因此,应严格执行种子进口规定。参考文献:(1) K. S. Bell等人,《欧洲植物病理学杂志》105:905,1999年。(2) C. D. Holevas等人,《贝纳基植物病理学研究所年报》19:52,2000年。(3) U. Merz,《欧洲和地中海植物保护组织通报》19:585,1989年。(4) M. E. Pantidou,希腊真菌-寄主索引。雅典贝纳基植物病理学研究所,1973年。