Park J-H, Seo S-T
Korea Forest Research Institute, Seoul 130-712, Republic of Korea.
Plant Dis. 2013 Jul;97(7):997. doi: 10.1094/PDIS-12-12-1167-PDN.
Smoothlip cymbidium (Cymbidium kanran Makino) is one of the national endangered species in Korea and in situ and ex situ conservation measures have been implemented for its threatened habitat and population. Jeju Island in Korea is one of few places where wild populations of the plant remain. In 2012, leaf blotch and blight occurred on leaves of smoothlip cymbidium in Jeju Island throughout the growing season. Affected plants were collected in May and October to determine the causal agent. Leaves had multiple brownish lesions that often developed from the leaf tip or leaf margin, eventually coalescing together and proceeding toward the base. In the lesions, fungal fruiting bodies formed concentric rings and exuded salmon-colored spore masses when put in moist chambers for 1 to 2 days. Colletotrichum gloeosporioides was consistently isolated from the spore masses or the margins of fresh lesions. When cultured on potato dextrose agar (PDA), colonies were olive to gray on the upper side and dark gray to black on the reverse side. On affected leaves, mature acervuli were dark brown to black, waxy, subepidermal, circular to ellipsoid, and 142 to 255 μm in diameter. Setae on the acervuli were dark brown, acicular, and 67 to 103 μm long. Conidia were hyaline, cylindrical with both ends rounded, and 13.7 to 19.2 × 4.0 to 6.1 μm. Hyphopodia were lobed and 7.6 to 13.7 × 5.7 to 11.6 μm (43 to 157 μm in area). These morphological characteristics were consistent with descriptions of C. gloeosporioides (1,3). The identities of two representative isolates were confirmed by sequencing the internal transcribed spacer (ITS) regions and the large subunit (LSU) rDNA (GenBank Accession Nos. KC408373, KC408374, and KC408375). BLAST analysis of the sequences from each isolate against the GenBank database found 99% similarities to more than 30 accessions of C. gloeosporioides (e.g., AY266392, EU552111). Pathogenicity was tested with one isolate on four leaves obtained from four plants of asymptomatic smoothlip cymbidium. Two places on the epidermis of each leaf were gently pinpricked using a sterile dissecting needle. One disk (0.6 cm diameter) of either PDA containing the fungus or sterile water agar was placed on one of the two places on each leaf. Laboratory film and aluminum foil held the disks in place. Inoculated leaves were kept in a moist chamber for 24 hr. After 1 week, buff-colored lesions with young acervuli were found on all fungus inoculation sites but no lesions occurred with control inoculations. C. gloeosporioides was recovered from all inoculations, but not from the controls. To our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on smoothlip cymbidium in Korea. The same fungus has been previously reported to cause anthracnose of other orchid plants in the genera Aspidistra, Cymbidium, and Dendrobium in Korea (2). This pathogen can pose a threat to smoothlip cymbidium. References: (1) M. Du et al. Mycologia 97: 641, 2005. (2) The Korean Society of Plant Pathology. List of Plant Diseases in Korea, 5th ed. The Korean Society of Plant Pathology, Suwon, Korea, 2009. (3) B. C. Sutton. Pages 1-26 in: Colletotrichum: Biology, Pathology, and Control. J. A. Bailey and M. J. Jeger, eds. CAB International, Wallingford, U.K., 1992.
春兰(Cymbidium kanran Makino)是韩国的国家濒危物种之一,针对其受到威胁的栖息地和种群已实施了原地和迁地保护措施。韩国济州岛是该植物野生种群留存的少数地区之一。2012年,济州岛的春兰在整个生长季节叶片上出现叶斑和枯萎病。在5月和10月采集受影响植株以确定致病因子。叶片上有多个褐色病斑,常从叶尖或叶缘开始形成,最终融合在一起并向基部蔓延。在病斑中,真菌子实体形成同心环,置于潮湿培养箱1至2天后会散发出鲑鱼色的孢子团。从孢子团或新鲜病斑边缘一直分离出胶孢炭疽菌(Colletotrichum gloeosporioides)。在马铃薯葡萄糖琼脂(PDA)上培养时,菌落上表面为橄榄色至灰色,下表面为深灰色至黑色。在受影响的叶片上,成熟的分生孢子盘深褐色至黑色,蜡质,位于表皮下,圆形至椭圆形,直径142至255μm。分生孢子盘上的刚毛深褐色,针状,长67至103μm。分生孢子透明,圆柱形,两端圆形,大小为13.7至19.2×4.0至6.1μm。附着胞呈叶状,大小为7.6至13.7×5.7至11.6μm(面积43至157μm)。这些形态特征与胶孢炭疽菌的描述一致(1,3)。通过对两个代表性分离株的内部转录间隔区(ITS)区域和大亚基(LSU)rDNA进行测序(GenBank登录号:KC408373、KC408374和KC408375),确认了它们的身份。将每个分离株的序列与GenBank数据库进行BLAST分析,发现与30多个胶孢炭疽菌的登录号有99%的相似性(例如AY266392、EU552111)。用一个分离株对从四株无症状春兰植株上获取的四片叶子进行致病性测试。用无菌解剖针在每片叶子表皮的两个部位轻轻扎孔。在每片叶子的两个部位之一放置一片含有该真菌的PDA或无菌水琼脂圆盘(直径0.6 cm)。用实验室薄膜和铝箔固定圆盘。接种后的叶子置于潮湿培养箱中24小时。1周后,在所有真菌接种部位发现有带有年轻分生孢子盘的浅黄色病斑,但对照接种部位未出现病斑。从所有接种部位重新分离出胶孢炭疽菌,但对照部位未分离到。据我们所知,这是韩国关于胶孢炭疽菌引起春兰炭疽病的首次报道。此前曾报道同一真菌在韩国引起蜘蛛抱蛋属、兰属和石斛属其他兰花植物的炭疽病(2)。这种病原菌会对春兰构成威胁。参考文献:(1)M. Du等人,《真菌学》97: 641, 2005。(2)韩国植物病理学会,《韩国植物病害名录》,第5版。韩国植物病理学会,韩国水原,2009。(3)B. C. Sutton,载于《炭疽菌:生物学、病理学与防治》,J. A. Bailey和M. J. Jeger编,CAB国际出版社,英国沃灵福德,1992年,第1 - 26页。