Michel V V, Hollenstein R, Stensvand A, Strømeng G M
Agroscope Changins-Wädenswil ACW, Research Center Conthey, CH-1964 Conthey, Switzerland.
Landwirtschaftliches Zentrum St. Gallen, CH-9230 Flawil, Switzerland.
Plant Dis. 2013 Sep;97(9):1246. doi: 10.1094/PDIS-08-12-0751-PDN.
Black elderberry (Sambucus nigra L.) is grown in Switzerland for flower and fruit production. Flowers are used for candy and syrup production, whereas the fruits are directly consumed as berries. In autumn 2008, the diagnostic laboratory of Agroscope ACW received a sample of strongly shriveled elderberry fruits from the extension office of the canton of St. Gallen. The sample originated from an experimental plot at Flawil, where 80% of the berries exhibited these symptoms. In years with high rainfall, infections of 100% of the berries can be observed in the production areas of Switzerland. Symptoms of anthracnose are only visible on the fruits, but not on the other plant organs. Berries start to shrivel when turning from green to black, and sporulation can be observed on ripe fruits under humid weather conditions. The sample was incubated in a moist chamber at room temperature, where it formed abundant acervuli producing salmon-colored spores at the fruit surface. Isolation from the acervuli on potato dextrose agar (PDA) containing an antibiotic (chlortetracycline) resulted in the growth of white to grey mycelium with salmon-colored spore masses. The reverse side of the PDA was red to violet. One-celled conidiospores were primarily fusiform, with an average size of 16.5 × 4 μm. Based on these morphological traits, the pathogen was previously identified as Colletotrichum acutatum J. H. Simmonds (2). A PCR using the primers CaInt2 and ITS4 (1) was run on a pure culture of the isolate from elderberry and confirmed this identification. A pathogenicity test was conducted from May to August 2010. The isolate from black elderberry and an isolate of C. acutatum from highbush blueberry (Vaccinium corymbosum) were multiplied separately on PDA on the laboratory bench (23 ± 2°C) for one week. Conidiospore suspensions of each isolate were prepared with 0.9% sterile NaCl solution and were adjusted to 1.2 × 10 spores/ml. Flower clusters of a single black elderberry tree at the Agroscope Research Center were inoculated at full flowering stage on May 26, 2010. Two sets of three healthy clusters were sprayed separately with the two spore suspensions until run-off. Spraying three healthy clusters with a sterile 0.9% NaCl solution served as control treatment. Immediately after inoculation, flower clusters were enclosed individually in transparent polyethylene bags for 2 days. To avoid excessive temperature inside the bags caused by solar radiation, the bagged flower clusters were placed below the leaves of the elderberry tree. During the 2 days, the average air temperature measured at the research center was 17 ± 2.5°C. Bags were removed and fruits of the treated clusters were harvested on July 27, 2010. Each cluster was incubated individually in a moist chamber on the laboratory bench (23 ± 2°C) for 10 days. Abundant formation of acervuli producing salmon-colored spores occurred on the fruits inoculated with either strain of C. acutatum. No such symptoms were produced on incubated fruits of the control treatment. From acervuli of the inoculated fruits, C. acutatum was reisolated on PDA. To our knowledge, this is the first report of C. acutatum on black elderberry. In Switzerland, a fungicide containing the active ingredient trifloxystrobine is registered to control C. acutatum on black elderberry. References: (1) S. K. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (2) B. C. Sutton. The Coelomycetes. CAB International, Wallingford, UK, 1980.
黑接骨木(Sambucus nigra L.)在瑞士种植用于生产花朵和果实。花朵用于制作糖果和糖浆,而果实则直接作为浆果食用。2008年秋季,瑞士农业研究中心(Agroscope ACW)诊断实验室收到来自圣加仑州推广办公室的一份严重皱缩的接骨木果实样本。该样本来自弗拉维尔的一个试验田,那里80%的浆果出现了这些症状。在瑞士降雨量较高的年份,在产区可观察到100%的浆果受到感染。炭疽病症状仅在果实上可见,在其他植物器官上则不明显。浆果从绿色变为黑色时开始皱缩,在潮湿天气条件下,成熟果实上可观察到产孢现象。将样本在室温的潮湿箱中培养,在果实表面形成了大量产生鲑鱼色孢子的分生孢子盘。在含有抗生素(金霉素)的马铃薯葡萄糖琼脂(PDA)上从分生孢子盘分离培养,长出白色至灰色的菌丝体,带有鲑鱼色的孢子团。PDA培养基的背面为红色至紫色。单细胞分生孢子主要呈梭形,平均大小为16.5×4μm。基于这些形态特征,该病原菌先前被鉴定为尖孢炭疽菌(Colletotrichum acutatum J. H. Simmonds)(2)。使用引物CaInt2和ITS4(1)对来自接骨木的分离菌株进行PCR扩增,证实了该鉴定结果。2010年5月至8月进行了致病性测试。将来自黑接骨木的分离菌株和来自高丛蓝莓(Vaccinium corymbosum)的尖孢炭疽菌分离菌株分别在实验室台面上(23±2°C)的PDA上培养一周。用0.9%无菌NaCl溶液制备每个分离菌株的分生孢子悬浮液,并将其浓度调整为1.2×10⁶个孢子/ml。2010年5月26日,在瑞士农业研究中心的一棵单株黑接骨木树的花簇处于盛花期时进行接种。两组,每组三个健康花簇,分别用两种孢子悬浮液喷雾直至径流。用无菌0.9%NaCl溶液喷雾三个健康花簇作为对照处理。接种后立即将花簇分别装入透明聚乙烯袋中2天。为避免袋内因太阳辐射导致温度过高,将套袋的花簇置于接骨木树的叶片下方。在这2天里,研究中心测得的平均气温为17±2.5°C。7月27日取下袋子,收获处理过的花簇上的果实。将每个花簇在实验室台面(23±2°C)的潮湿箱中单独培养10天。接种任何一种尖孢炭疽菌菌株的果实上都大量形成了产生鲑鱼色孢子的分生孢子盘。对照处理培养的果实未出现此类症状。从接种果实的分生孢子盘中,在PDA上再次分离出尖孢炭疽菌。据我们所知,这是尖孢炭疽菌在黑接骨木上的首次报道。在瑞士,一种含有活性成分肟菌酯的杀菌剂已注册用于防治黑接骨木上的尖孢炭疽菌。参考文献:(1)S. K. Sreenivasaprasad等人,《植物病理学》45:650,1996年。(2)B. C. Sutton,《腔孢菌纲》,CAB国际出版社,英国沃灵福德,1980年。