Williamson S M, Sutton T B
Department of Plant Pathology, North Carolina State University, Raleigh 27695.
Plant Dis. 2010 May;94(5):634. doi: 10.1094/PDIS-94-5-0634A.
Persimmon trees are important for their fruit as well as their colorful fruit and foliage in the fall. Persimmon fruit (Japanese persimmon, Diospyros kaki cv. Fuyu) were collected in November 2008 from a tree in Windsor, NC, located in the Coastal Plain. Fruit were not symptomatic on the tree but developed dark lesions after harvest. Isolations from six fruit yielded seven isolates of Colletotrichum acutatum J. H. Simmonds. After incubation at 25°C under continuous light for 15 days on potato dextrose agar (PDA), all isolates had gray aerial mycelium, but the inverse sides of the plates of six isolates were maroon and one was beige. Masses of salmon-colored conidia were formed first in the center of the colonies, then were observed scattered across the colonies in older cultures. Conidia were hyaline, one-celled, elliptic with one or both ends pointed, and measured 8.1 to 16.3 × 3.1 to 5 μm. Setae and sclerotia were not observed. There were also dark structures measuring 1 to 10 mm that were partially embedded in the agar that contained conidia. Cultural and conidial characteristics of the isolates were similar to those of C. acutatum (3). PCR amplification was performed with the species-specific primer pair CaInt2/ITS4 (2) and genomic DNA from the original isolates and isolates obtained from inoculated fruit. An amplification product of approximately 490 bp, which is specific for C. acutatum, was observed. To fulfill Koch's postulates, persimmon fruit obtained from the grocery store were surface disinfested with 0.5% sodium hypochlorite and sterile filter paper disks dipped in conidial suspensions (1 × 10 conidia/ml) of two C. acutatum isolates (maroon and beige reverse) or sterile, deionized water were placed on the fruit. Three fruit were inoculated per treatment and the disks were placed on four locations on each fruit. Parafilm was wrapped around the diameter of the fruit to keep the filter paper disks moist and in place. Fruit were placed in moist chambers and incubated at 25°C. After 3 days, the Parafilm was removed and the fruit returned to the moist chambers. Small, dark lesions were observed on fruit inoculated with each isolate of C. acutatum when the filter paper disks were removed. Ten days after inoculation, dark lesions and acervuli with salmon-colored masses of conidia were observed on fruit inoculated with both isolates of C. acutatum and the fruit were soft. After 12 days, there were abundant masses of conidia and the inoculated areas were decayed. Control fruit remained firm and did not develop symptoms. Cultures obtained from the fruit and the conidia produced were typical of the isolates used to inoculate the fruit. C. acutatum has been reported to cause fruit rot on persimmon fruit in New Zealand (1). To our knowledge, this is the first report of C. acutatum on persimmon fruit in the United States. References: (1) R. Lardner et al. Mycol. Res. 103:275, 1999. (2) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (3) B. C. Sutton. Page 523 in: Coelomycetes. Commonwealth Agricultural Bureaux, Great Britain. 1980.
柿树因其果实以及秋季色彩斑斓的果实和树叶而显得重要。2008年11月,从北卡罗来纳州温莎市沿海平原的一棵树上采集了柿果(日本柿,柿属富岳品种)。这些果实在树上时没有症状,但收获后出现了深色病斑。从六个果实上分离得到了七株尖孢炭疽菌(J. H. Simmonds)。在马铃薯葡萄糖琼脂(PDA)上于25°C连续光照下培养15天后,所有分离株都有灰色气生菌丝体,但六个分离株平板的反面是栗色,一个是米色。鲑鱼色分生孢子团首先在菌落中心形成,然后在较老的培养物中分散在整个菌落中观察到。分生孢子透明,单细胞,椭圆形,一端或两端尖,大小为8.1至16.3×3.1至5μm。未观察到刚毛和菌核。在含有分生孢子的琼脂中还观察到部分埋入的1至10毫米的深色结构。分离株的培养和分生孢子特征与尖孢炭疽菌相似(3)。使用物种特异性引物对CaInt2/ITS4(2)以及原始分离株和接种果实后获得的分离株的基因组DNA进行PCR扩增。观察到了约为尖孢炭疽菌特异性的490bp扩增产物。为了验证科赫法则(Koch's postulates),从杂货店获得的柿果用0.5%次氯酸钠进行表面消毒,将浸有两株尖孢炭疽菌分离株(栗色和米色反面)分生孢子悬浮液(1×10分生孢子/ml)的无菌滤纸圆盘或无菌去离子水放在果实上。每个处理接种三个果实,圆盘放在每个果实的四个位置。用保鲜膜包裹果实直径以保持滤纸圆盘湿润并固定在位。将果实置于保湿箱中,在25°C下培养。3天后,取下保鲜膜,将果实放回保湿箱。当取下滤纸圆盘时,在接种了每株尖孢炭疽菌分离株的果实上观察到小的深色病斑。接种10天后,在接种了两株尖孢炭疽菌分离株的果实上观察到深色病斑和带有鲑鱼色分生孢子团的分生孢子盘,果实变软。12天后,有大量分生孢子团,接种区域腐烂。对照果实保持坚硬,未出现症状。从果实上获得的培养物和产生的分生孢子与用于接种果实的分离株典型特征一致。据报道,尖孢炭疽菌在新西兰会导致柿果腐烂(1)。据我们所知,这是美国关于尖孢炭疽菌导致柿果发病情况的首次报道。参考文献:(1)R. Lardner等人,《真菌学研究》103:275,1999年。(2)S. Sreenivasaprasad等人,《植物病理学》45:650,1996年。(3)B.C. Sutton,载于《腔孢纲》第523页。英国联邦农业局,1980年。