Li Q L, Mo J Y, Huang S P, Guo T X, Pan Z B, Ning P, Hsiang T
Institute of Plant Protection, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi, 530007, China, and Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests, Nanning, Guangxi, 530007, China.
Department of Biotechnology, Guangxi Agricultural Vocation-Technical College, Nanning, Guangxi, 530007, China.
Plant Dis. 2013 Oct;97(10):1383. doi: 10.1094/PDIS-03-13-0346-PDN.
Lobelia chinensis is a perennial herbaceous plant in the family Campanulaceae that is native to China, where it grows well in moist to wet soils. It is commonly used as a Chinese herbal medicine. In May 2012, symptoms of leaf spot were observed on leaves of L. chinensis in Nanning, Guangxi Zhuang Autonomous Region, China. The leaf lesions began as small, water-soaked, pale greenish to grayish spots, which enlarged to gray to pale yellowish spots, 4 to 6 mm in diameter. At later stages, numerous acervuli appeared on the lesions. Acervuli were mostly epiphyllous, and 40 to 196 μm in diameter. On potato dextrose agar (PDA), a fungus was consistently recovered from symptomatic leaf samples, with a 93% isolation rate from 60 leaf pieces that were surface sterilized in 75% ethanol for 30 s and then in 0.1% mercuric chloride for 45 s. Three single-spore isolates were used to evaluate cultural and morphological characteristics of the pathogen. Setae were two to three septate, dark brown at the base, acicular, and up to 90 μm long. Conidia were long oblong-elliptical, guttulate, hyaline, and 11 to 20 × 4.1 to 6.3 μm (mean 15.2 × 5.1 μm). These morphological characteristics of the fungus were consistent with the description of Colletotrichum magna (teleomorph Glomerella magna Jenkins & Winstead) (1). The rDNA internal transcribed spacer (ITS) region of one isolate, LC-1, was sequenced (GenBank Accession No. KC815123), and it showed 100% identity to G. magna, GenBank HM163187.1, an isolate from Brazil cultured from papaya (2). Although KC815123 was identified as G. magna, it shows 99% identity to GenBank sequences from isolates of C. magna, and more research is needed to elucidate the relationships between these taxa, especially with consideration to host specificity. Pathogenicity tests were performed with each of the three isolates by spraying conidial suspensions (1 × 10 conidia/ml) containing 0.1% Tween 20 onto the surfaces of leaves of 30-day-old and 6- to 8-cm-high plants. For each isolate, 30 leaves from five replicate plants were treated. Control plants were treated with sterilized water containing 0.1% Tween 20. All plants were incubated for 36 h at 25°C and 90% relative humidity in an artificial climate chamber, and then moved into a greenhouse. Seven days after inoculation, gray spots typical of field symptoms were observed on all inoculated leaves, but no symptoms were seen on water-treated control plants. Koch's postulates were fulfilled by reisolation of G. magna from diseased leaves. To our knowledge, this is the first report of G. magna infecting L. chinensis worldwide. References: (1) M. Z. Du et al. Mycologia 97:641, 2005. (2) R. J. Nascimento et al. Plant Dis. 94:1506, 2010.
半边莲是桔梗科的一种多年生草本植物,原产于中国,在湿润至潮湿的土壤中生长良好。它通常被用作一种中草药。2012年5月,在中国广西壮族自治区南宁市的半边莲叶片上观察到叶斑病症状。叶片病斑最初为小的、水渍状、淡绿色至灰色斑点,随后扩大为灰色至淡黄色斑点,直径4至6毫米。在后期阶段,病斑上出现大量分生孢子盘。分生孢子盘大多生在叶面上,直径40至196微米。在马铃薯葡萄糖琼脂(PDA)培养基上,从有症状的叶片样本中始终能分离出一种真菌,从60片经75%乙醇表面消毒30秒、再用0.1%氯化汞消毒45秒的叶片中,分离率为93%。使用三个单孢分离株来评估病原菌的培养和形态特征。刚毛具2至3个隔膜,基部深褐色,针状,长达90微米。分生孢子长椭圆形至椭圆形,具油滴,无色透明,大小为11至20×4.1至6.3微米(平均15.2×5.1微米)。该真菌的这些形态特征与大炭疽菌(有性型为大光壳炭疽菌Jenkins & Winstead)的描述一致(1)。对其中一个分离株LC - 1的核糖体DNA内转录间隔区(ITS)进行了测序(GenBank登录号KC815123),结果显示它与大光壳炭疽菌(GenBank登录号HM163187.1,从巴西番木瓜上分离培养得到的一个分离株)的序列一致性为100%(2)。尽管KC815123被鉴定为大光壳炭疽菌,但它与大炭疽菌分离株的GenBank序列一致性为99%,需要更多研究来阐明这些分类单元之间的关系,尤其是考虑到寄主特异性。用三个分离株中的每一个进行致病性测试,将含有0.1%吐温20的分生孢子悬浮液(1×10个分生孢子/毫升)喷洒到30日龄、高6至8厘米的植株叶片表面。对于每个分离株,处理来自五株重复植株的30片叶子。对照植株用含有0.1%吐温20的无菌水进行处理。所有植株在人工气候箱中于25°C和90%相对湿度下培养36小时,然后转移到温室中。接种七天后,在所有接种叶片上观察到典型的田间症状灰色斑点,但水处理的对照植株上未出现症状。通过从病叶中重新分离出大光壳炭疽菌,柯赫氏法则得到了验证。据我们所知,这是大光壳炭疽菌侵染半边莲在全球范围内的首次报道。参考文献:(1)M. Z. Du等人,《真菌学》97:641,2005年。(2)R. J. Nascimento等人,《植物病害》94:1506,2010年。