Diao Y Z, Zhang C, Lin D, Liu X L
Department of Plant Pathology, China Agricultural University, Beijing 100193, China.
Plant Dis. 2014 May;98(5):687. doi: 10.1094/PDIS-05-13-0491-PDN.
Colletotrichum truncatum (syn. C. capsici) is an important plant pathogen that has a wide host range including pepper, eggplant, muskmelon, chickpea, grapes, and many other species of plants (1,2). Anthracnose fruit rot incited by C. coccodes is a prevalent disease in some major tomato (Solanum esculentum) producing regions in China. In October 2012, anthracnose symptoms (circular, sunken lesions with spore masses produced in black acervuli) were observed on ripe tomato fruit in Fuzhou City of Fujian Province, China. Three single-spore isolates (FZ-1, FZ-2, and FZ-3) were derived from fungal cultures isolated from different infected fruits from the same farm. A mycelial plug (5 mm in diameter) from the growing edge of an active colony of each isolate was transferred onto potato dextrose agar (PDA) and incubated at 28°C. Colonies grown on PDA changed from grayish to dark grey with an average colony diameter of 71.8 mm after 7 days. Conidia were falcate and 17.6 to 21.6 × 2.57 to 3.31 μm. Growth rate measured by colony diameter was greater at 25 to 30°C than at other temperatures tested (12, 18, 20, and 37°C). Based upon these morphological and cultural characteristics, the causal agent was identified as C. truncatum (3). To test the three isolates for pathogenicity, detached ripe tomato fruits were each inoculated with 1 μl of a conidial suspension (10 conidia/ml) using either injection or applying a droplet of the spore suspension on the surface of each fruit; the control treatment consisted of fruit that was treated with 1 μl of sterilized water using the two methods of inoculation mentioned above. Five replicate ripe fruits were inoculated with each of the three isolates using each method described above, and incubated in a moist chamber at 25°C. Five ripe fruits were used the negative control treatment for each inoculation method. After 7 days, typical anthracnose symptoms had developed on the inoculated fruit but not on control fruit. To confirm identity of the three isolates, the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA), the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, a partial sequence of the actin (ACT) gene, and partial histone 3 (HIS3) gene were amplified with ITS1 and ITS4 universal primers, GDF1 and GDR1 primers, ACT-512F and ACT-783R primers, and CYLH3F and CYLH3R primers, respectively (1). The ITS consensus sequence (Accession No. KC460308) of these isolates shared 99% homology with the ITS sequence of C. truncatum in GenBank (AJ301945), and the three other consensus sequences were all 100% homologous with the corresponding sequences of C. truncatum in Genbank (GU228254, GU227960, and GU228058, respectively). The pathogen was re-isolated from the inoculated fruit but not from the control fruit, and the identity of the re-isolates was confirmed to be C. truncatum by morphological features and based on the ITS, GAPDH, ACT, and HIS3 sequences as described above. To our knowledge, this is the first report of C. truncatum causing anthracnose on tomato in China. References: (1) U. Damm et al. Fungal Divers. 39:45, 2009. (2) K. K. Pandey. J. Mycol. Plant Pathol. 36:104, 2006. (3) I. S. Sawant et al. New Dis. Rep. 25:2, 2012.
菜豆炭疽菌(同义词:辣椒炭疽菌)是一种重要的植物病原菌,寄主范围广泛,包括辣椒、茄子、甜瓜、鹰嘴豆、葡萄及许多其他植物物种(1,2)。由球炭疽菌引起的炭疽病果腐是中国一些主要番茄(茄属)产区的一种常见病害。2012年10月,在中国福建省福州市,成熟番茄果实上观察到炭疽病症状(圆形、凹陷病斑,在黑色分生孢子盘上产生孢子堆)。从同一农场不同感染果实分离得到的真菌培养物中获得了三个单孢分离株(FZ - 1、FZ - 2和FZ - 3)。将每个分离株活跃菌落生长边缘的一个菌丝块(直径5毫米)转移到马铃薯葡萄糖琼脂(PDA)上,在28°C下培养。在PDA上生长的菌落从浅灰色变为深灰色,7天后平均菌落直径为71.8毫米。分生孢子镰刀形,大小为17.6至21.6×2.57至3.31微米。通过菌落直径测量的生长速率在25至30°C时比其他测试温度(12、18、20和37°C)下更高。基于这些形态和培养特征,致病因子被鉴定为菜豆炭疽菌(3)。为测试这三个分离株的致病性,使用注射或在每个果实表面滴加一滴孢子悬浮液的方法,将1微升分生孢子悬浮液(10个分生孢子/毫升)接种到离体成熟番茄果实上;对照处理包括使用上述两种接种方法用1微升无菌水处理的果实。使用上述每种方法,用这三个分离株中的每一个接种五个重复的成熟果实,并在25°C的保湿箱中培养。对于每种接种方法,使用五个成熟果实作为阴性对照处理。7天后,接种果实上出现了典型的炭疽病症状,而对照果实上没有。为确认这三个分离株的身份,分别用ITS1和ITS4通用引物、GDF1和GDR1引物、ACT - 512F和ACT - 783R引物以及CYLH3F和CYLH3R引物扩增核糖体DNA(rDNA)的内部转录间隔区(ITS)区域、甘油醛 - 3 - 磷酸脱氢酶(GAPDH)基因、肌动蛋白(ACT)基因的部分序列和部分组蛋白3(HIS3)基因(1)。这些分离株的ITS一致序列(登录号KC460308)与GenBank中菜豆炭疽菌的ITS序列(AJ301945)具有99%的同源性,其他三个一致序列分别与Genbank中菜豆炭疽菌的相应序列(GU228254、GU227960和GU228058)具有100%的同源性。病原菌从接种果实中重新分离得到,但未从对照果实中分离得到,通过形态特征以及基于上述ITS、GAPDH、ACT和HIS3序列,重新分离株的身份被确认为菜豆炭疽菌。据我们所知,这是菜豆炭疽菌在中国引起番茄炭疽病的首次报道。参考文献:(1)U. Damm等人,真菌多样性。39:45,2009。(2)K. K. Pandey,真菌学与植物病理学杂志。36:104,2006。(3)I. S. Sawant等人,新病害报告。25:2,2012。