Kim Y K, Caiazzo R, Sikdar P, Xiao C L
Pace International, Wapato, WA.
Washington State University, TFREC, Wenatchee.
Plant Dis. 2013 Sep;97(9):1257. doi: 10.1094/PDIS-03-13-0320-PDN.
In March 2012, decayed 'Empire' apple fruit (Malus × domestica Borkh.) were sampled from apples grown in Albion (Orleans County) in New York State and stored in bins for 6 months under controlled atmosphere at a commercial packinghouse. At the packinghouse following storage prior to be packed, the fruit were completely rotten, spongy to firm, and light brown without pycnidia. All fruit rots originated from either stem-end or calyx-end infections but no wound infections were observed. The incidence of fruit with these symptoms in the total decay was relatively low (0.1%). To isolate the causal agent, small fragments of fruit flesh from 12 decayed fruit were cut and placed on potato dextrose agar (PDA) acidified with 0.1% lactic acid. The plates were incubated at 20°C for 4 days and sub-cultured on PDA to obtain a pure culture. The colonies initially appeared with dense hyaline mycelium and later turned light yellow to yellow, and black pycnidia formed after about 2 weeks of incubation under a 24-h fluorescent light at 20°C. Conidia were light brown to brown, clavate to subglobose to irregular, and 15 × 8 μm on average. The fungus was identified as Sphaeropsis pyriputrescens Xiao & J.D. Rogers based on the morphology of the fungus (3). The identity of a representative isolate was further confirmed by analysis of nucleotide sequences of the internal transcribed spacer (ITS) regions amplified using the primers ITS1/ITS4. A BLAST search in GenBank showed that the sequence had 99% homology to an S. pyriputrescens sequence (Accession No. GQ374241). One representative isolate was tested for pathogenicity on apple fruit. Organic 'Red Delicious' apple fruit were surface-disinfected in 0.6% sodium hypochlorite solution for 5 min, rinsed twice with deionized water, and air-dried. Each fruit was wounded with a sterilized finish-nail head (3 mm in depth and 4 mm in diameter) and inoculated by placing a 4-mm-diameter plug from the leading edge of a 4-day-old PDA culture on the wound. Control fruit were treated with sterile PDA plugs. The inoculation site was covered with two layers of moist cheesecloth to avoid dehydration. There were four 10-fruit replicates for each treatment, and fruit were placed in plastic crispers and stored at 4°C for 4 weeks. The experiments were conducted twice. Sphaeropsis rot developed on all inoculated fruit, while no decays appeared on the control fruit. Koch's postulates were fulfilled by reisolating the fungus from the decayed fruit. Sphaeropsis rot is a recently reported postharvest fruit rot disease of apple and pear (1,3). The disease was first observed on 'd'Anjou' pears, and later more serious economic losses were observed in apples in Washington State (1). The disease has also since been reported in British Columbia, Canada (2). To the best of our knowledge, this is the first report of the occurrence of Sphaeropsis rot caused by S. pyriputrescens on apple in New York or in any region outside of the Pacific Northwest in North America. References: (1) Y. K. Kim and C. L. Xiao. Plant Dis. 92:940, 2008. (2) P. L. Sholberg et al. Plant Dis. 93:843, 2009. (3) C. L. Xiao et al. Plant Dis. 88:223, 2004.
2012年3月,从纽约州奥尔良县阿尔比恩种植并在商业包装厂气调贮藏6个月的“帝国”苹果上采集了腐烂果实样本。在包装厂贮藏后包装前,果实已完全腐烂,质地从松软到坚硬,呈浅褐色,无分生孢子器。所有果实腐烂均源于果梗端或萼端感染,未观察到伤口感染。出现这些症状的果实占总腐烂果实的比例相对较低(0.1%)。为分离致病因子,从12个腐烂果实上切取小块果肉,置于添加0.1%乳酸酸化的马铃薯葡萄糖琼脂(PDA)上。平板在20°C培养4天,然后转接至PDA上以获得纯培养物。菌落最初呈现浓密的透明菌丝体,随后变为浅黄色至黄色,在20°C、24小时荧光灯下培养约2周后形成黑色分生孢子器。分生孢子浅褐色至褐色,棍棒状至近球形至不规则形,平均大小为15×8μm。根据真菌形态,该真菌被鉴定为梨球壳孢(Sphaeropsis pyriputrescens Xiao & J.D. Rogers)(3)。通过对使用引物ITS1/ITS4扩增的内部转录间隔区(ITS)核苷酸序列进行分析,进一步确认了一个代表性分离株的身份。在GenBank中进行的BLAST搜索显示,该序列与梨球壳孢序列(登录号GQ374241)有99%的同源性。对一个代表性分离株进行了苹果果实致病性测试。将有机“红地厘蛇果”果实置于0.6%次氯酸钠溶液中表面消毒5分钟,用去离子水冲洗两次,然后风干。每个果实用灭菌的鞋钉扎伤(深度3mm,直径4mm),并通过在伤口处放置一块取自4日龄PDA培养物前沿的直径4mm的菌块进行接种。对照果实用无菌PDA菌块处理。接种部位用两层湿纱布覆盖以避免脱水。每个处理有4个重复,每个重复10个果实,果实置于塑料保鲜盒中,在4°C下贮藏4周。试验进行了两次。所有接种果实均发生了梨球壳孢腐烂,而对照果实未出现腐烂。从腐烂果实中重新分离出该真菌,从而证实了柯赫氏法则。梨球壳孢腐烂是一种最近报道的苹果和梨采后果实腐烂病(1,3)。该病最初在“安久”梨上被观察到,后来在华盛顿州的苹果上造成了更严重的经济损失(1)。此后,加拿大不列颠哥伦比亚省也报道了该病(2)。据我们所知,这是北美纽约州或太平洋西北地区以外任何地区首次报道由梨球壳孢引起的苹果梨球壳孢腐烂病的发生。参考文献:(1)Y.K. Kim和C.L. Xiao。植物病害92:940,2008。(2)P.L. Sholberg等人。植物病害93:843,2009。(3)C.L. Xiao等人。植物病害88:223,2004。