Novotný D, Křížková I, Krátká J, Salava J
Crop Research Institute, Drnovská 507, 161 06, Prague 6, Czech Republic. This study was supported by project MZE0002700603 from the Ministry of Agriculture of the Czech Republic.
Plant Dis. 2007 Nov;91(11):1516. doi: 10.1094/PDIS-91-11-1516B.
Colletotrichum acutatum J. H. Simmond is the causal agent of anthracnose rot and strawberry blackspot. This pathogen is listed by the EPPO as a regulated (formerly quarantined) organism for all European countries and is widely distributed throughout Europe (e.g., the United Kingdom, France, and Germany) (2). In the autumn of 2005, typical symptoms of anthracnose caused by C. acutatum (circular, dark, and sunken spots on fruit, dark, sunken lesions on petioles, and withering of the leaf, buds, and flowers) were repeatedly observed on field-grown strawberry plants in the Mělník Region of central Bohemia and Břeclav Region in southern Moravia, Czech Republic. Strawberry fruits and petioles showing typical symptoms were surface sterilized (30 s in 70% ethanol, 1 min in 10% NaOCl, and 15 s in 70% ethanol), rinsed in sterile water, dissected under aseptic conditions, and plated on 2% malt extract agar or placed in wet chambers and incubated at room temperature (18 to 20°C) for 10 days. All isolated strains were independently identified by morphological characteristics, plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) (1), and PCR with the C. acutatum-specific primers ITS4 and CaInt2 (3). Morphological studies of C. acutatum were carried out on potato dextrose agar (4). The colonies were white, cream, grayish, or rose-orange before sporulation and the colony reverse was cream to orange with brown spots. The mycelial growth rate was 7.5 mm per day at 25°C. The conidia were one-celled, hyaline, cylindrical, 11.3 to 19.7 × 3.6 to 5.5 μm, and the majority of conidia were pointed at either or both ends. The appresoria were brown, globose to ellipsoidal, 5.0 to 7.5 × 5.0 to 6.2 μm, and the sclerotia were absent. Ten strawberry plants with green fruits of each cultivar Elsanta and Kama were sprayed with 500 ml of suspension of C. acutatum conidia (10 conidia per ml). This test was carried out in the glasshouse under quarantine conditions at 20 to 25°C. C. acutatum caused withering of the flowers or dark brown spots on green fruits on five plants of cv. Elsanta and on four plants of cv. Kama after a 6-week incubation period. Isolation and identification of the pathogen from the diseased tissues were done as described above. C. acutatum was reisolated from three fruits, four leaf blades, and four petioles from five plants of cv. Elsanta and four fruits, four leaf blades, and two petioles from four plants of cv. Kama. The fungus was not reisolated from the control strawberry plants. In three cases, the pathogen was detected in the crown of plants of cv. Elsanta by PCR and ELISA. References: (1) Anonymous. OEPP/EPPO Bull. 34:155, 2004. (2) I. M. Smith and L. M. F. Charles, eds. Distribution Maps of Quarantine Pests for the European Union and for the European and Mediterranean Plant Protection Organization. CABI Publishing, Wallingford, UK, 1998. (3) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (4) P. Talhinhas et al. Phytopathology 92:986, 2002.
尖孢炭疽菌(J. H. Simmond)是炭疽病腐烂和草莓黑斑病的病原菌。该病原菌被欧洲和地中海植物保护组织(EPPO)列为所有欧洲国家的管制(以前为检疫)生物,在欧洲广泛分布(如英国、法国和德国)(2)。2005年秋季,在捷克共和国中波希米亚州的梅尔尼克地区和南摩拉维亚州的布雷clav地区,在田间种植的草莓植株上反复观察到由尖孢炭疽菌引起的典型炭疽病症状(果实上出现圆形、深色和凹陷斑点,叶柄上出现深色、凹陷病斑,叶片、芽和花朵枯萎)。对表现出典型症状的草莓果实和叶柄进行表面消毒(在70%乙醇中浸泡30秒,在10%次氯酸钠中浸泡1分钟,在70%乙醇中浸泡15秒),用无菌水冲洗,在无菌条件下解剖,接种在2%麦芽提取物琼脂上或置于湿盒中,在室温(18至20°C)下培养10天。所有分离菌株均通过形态特征、平板捕获抗原酶联免疫吸附测定(PTA - ELISA)(1)以及使用尖孢炭疽菌特异性引物ITS4和CaInt2的PCR(3)进行独立鉴定。尖孢炭疽菌的形态学研究在马铃薯葡萄糖琼脂上进行(4)。在产孢前,菌落为白色、奶油色、灰色或玫瑰橙色、菌落背面为奶油色至橙色并带有褐色斑点。在25°C下,菌丝生长速率为每天7.5毫米。分生孢子单细胞、无色、圆柱形,11.3至19.7×3.6至5.5μm,大多数分生孢子一端或两端尖锐。附着胞褐色,球形至椭圆形,5.0至7.5×5.0至6.2μm,无菌核。对每个品种Elsanta和Kama的10株带有绿色果实的草莓植株喷洒500毫升尖孢炭疽菌分生孢子悬浮液(每毫升10个分生孢子)。该试验在温室中于20至25°C的检疫条件下进行。经过6周的培养期后,尖孢炭疽菌在Elsanta品种的5株植株上导致花朵枯萎或绿色果实上出现深褐色斑点,在Kama品种的4株植株上出现同样症状。从患病组织中分离和鉴定病原菌的方法如上所述。从Elsanta品种的5株植株的3个果实、4片叶片和4个叶柄以及Kama品种的4株植株的4个果实、4片叶片和2个叶柄中再次分离出尖孢炭疽菌。在对照草莓植株上未再次分离到该真菌。在3个案例中,通过PCR和ELISA在Elsanta品种植株的根茎中检测到病原菌。参考文献:(1)匿名。OEPP/EPPO通报34:155,2004。(2)I. M. 史密斯和L. M. F. 查尔斯编。欧盟及欧洲和地中海植物保护组织检疫性有害生物分布图。CABI出版社,英国沃灵福德,1998。(3)S. Sreenivasaprasad等人。植物病理学45:650,1996。(4)P. Talhinhas等人。植物病理学92:986,2002。