Yin L F, Chen S N, Cai M L, Li G Q, Luo C X
College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
College of Plant Science and Technology and the Key Lab of Crop Disease Monitoring and Safety Control in Hubei Province, Huazhong Agricultural University, Wuhan 430070, China.
Plant Dis. 2014 May;98(5):694. doi: 10.1094/PDIS-09-13-0995-PDN.
In May 2013, apricot (Prunus armeniaca) fruits covered with grayish, conidial masses were collected from an unknown cultivar in an experimental field of Huazhong Agricultural University, Wuhan, Hubei Province. About 3 to 5% of fruit was infected and affected apricots had tan to white zones of sporulation, which resembled brown rot caused by Monilia species. Conidia were harvested from the surface of the sporulating apricot fruit and spread onto shallow potato dextrose agar (PDA) media (about 2 mm in thickness) using sterile cotton swabs. Conidia were lemon-shaped and mean size was 15.7 (10 to 22.5) × 25 (16.25 to 35) μm. Conidia on PDA were incubated at 23°C for 3 h in darkness, then observed under microscope. More than two germ tubes were produced from each conidium, which was the distinctive trait of Monilia mumecola species (2). Single-spore isolates were obtained and 3 isolates were cultured on PDA in petri dishes. Mycelium grew at an average of 15 mm per day, and the colony showed concentric rings of mycelium with lobbed margins at 23°C in darkness. A 712-bp fragment was PCR amplified from β-tubulin gene (TUB2) of all the nine isolates investigated indicative of M. mumecola (2). The ribosomal ITS1-5.8S-ITS2 regions of nine isolates were also PCR-amplified from genomic DNA using primers ITS1 and ITS4 and then sequenced (4). ITS sequences were identical to ITS sequences of M. mumecola from China (HQ908786) and Japan (AB125613, AB125614, and AB125620), but only has 98% and 97% identity with the closest species M. laxa (EU042149) and M. fructicola (HQ908789) according to BLAST search in GenBank. Pathogenicity was confirmed by inoculating mycelial plugs of three isolates into six surface-sterilized apricots wounded with a 6-mm diameter sterile cork borer. Control fruit received plain PDA plugs and was incubated in a moist chamber at 23°C with 12 h light/12 h dark. All inoculated fruit developed typical brown rot symptoms with sporulating areas as described above after 3 days of incubation, while control fruits remained healthy. The developing spores on inoculated fruit were re-isolated and confirmed to be M. mumecola. M. mumecola was first isolated from Prunus mume in Japan in 1982 as an unknown Monilia species (3), then identified and the nomenclature was provided in 2004 (1). To our knowledge, this is the first report of M. mumecola on P. armeniaca indicating that M. mumecola has spread to different hosts. References: (1) Y. Harada et al. J. Gen. Plant Pathol. 70:297, 2004. (2) M. J. Hu et al. Plos One, 6(9):e24990, 2011. (3) S. Nakao. Kongetsu-no-noyaku, 1:92, 1992. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.
2013年5月,从湖北省武汉市华中农业大学实验田的一个未知品种杏树上采集到表面覆盖着灰色分生孢子团的杏果。约3%至5%的果实被感染,受影响的杏子有棕褐色至白色的产孢区域,类似由链核盘菌属物种引起的褐腐病。从产孢杏果表面收集分生孢子,并用无菌棉签将其涂抹在浅马铃薯葡萄糖琼脂(PDA)培养基(厚度约2毫米)上。分生孢子呈柠檬形,平均大小为15.7(10至22.5)×25(16.25至35)微米。PDA上的分生孢子在23°C黑暗条件下培养3小时,然后在显微镜下观察。每个分生孢子产生两根以上芽管,这是梅链核盘菌的显著特征(2)。获得单孢分离物,并在培养皿中的PDA上培养3个分离物。菌丝体平均每天生长15毫米,菌落显示出同心的菌丝环,在23°C黑暗条件下边缘呈叶状。从所研究的所有9个分离物的β-微管蛋白基因(TUB2)中PCR扩增出一个712bp的片段,表明为梅链核盘菌(2)。还使用引物ITS1和ITS4从基因组DNA中PCR扩增9个分离物的核糖体ITS1-5.8S-ITS2区域,然后进行测序(4)。ITS序列与来自中国(HQ908786)和日本(AB125613、AB125614和AB125620)的梅链核盘菌的ITS序列相同,但根据GenBank中的BLAST搜索,与最接近的种松弛链核盘菌(EU042149)和果生链核盘菌(HQ908789)的同一性仅为98%和97%。通过将3个分离物的菌丝块接种到6个用直径6毫米的无菌软木钻打孔并表面消毒的杏子中,证实了致病性。对照果实接种普通PDA块,并在23°C、12小时光照/12小时黑暗的潮湿箱中培养。接种后3天,所有接种的果实都出现了上述典型的褐腐症状和产孢区域,而对照果实保持健康。对接种果实上发育的孢子进行再分离,证实为梅链核盘菌。梅链核盘菌于1982年在日本首次从梅中分离出来,当时为一种未知的链核盘菌属物种(3),随后于2004年被鉴定并给出了命名(1)。据我们所知,这是梅链核盘菌在杏树上的首次报道,表明梅链核盘菌已传播到不同寄主。参考文献:(1)Y. Harada等人,《植物病理学报》70:297,2004年。(2)M. J. Hu等人,《公共科学图书馆·综合》6(9):e24990,2011年。(3)S. Nakao,《昆虫月刊》1:92,1992年。(4)T. J. White等人,载于《PCR实验方案:方法与应用指南》第315页。M. A. Innis等人编。学术出版社,圣地亚哥,1990年。