Jelev Z J, Bobev S G, Minz D, Maymon M, Freeman S
Agricultural University, 4000 Plovdiv, Bulgaria.
Institute for Soil, Water and Environmental Sciences, ARO, Volcani Research Center, Bet Dagan 50-250 Israel.
Plant Dis. 2008 Jan;92(1):172. doi: 10.1094/PDIS-92-1-0172C.
In the late summer of 2005, sporadic and unusual damage was observed on pepper (Capsicum annuum cv. Kurtovska kapia and local cv. Ribka) on two farms and tomato (Lycopersicon esculentum cv. Florida 47) fruits on one farm in the Plovdiv Region of Bulgaria. Dry, round, sunken zones (10 to 20 mm) were observed on pepper fruits that preserved their natural skin color even after black acervuli containing orange masses of conidia appeared. Eventually, the lesions turned brown, coalesced, and the fruits mummified on the plants. Tomato fruits developed similar symptoms, with less prominent discoloration and fewer acervuli. The pathogen was easily isolated from both hosts on potato dextrose agar where it formed white-to-gray colonies with salmon orange pigmentation on the reverse side of the plates. Conidia that formed were hyaline, fusiform, aseptate, and measured 13.3 to 17.4 × 3.5 to 5.5 μm and 11.6 to 15.5 × 4.1 to 5.0 μm for pepper and tomato isolates, respectively. Both cultural and morphological characteristics of the isolates were similar to those described for Colletotrichum acutatum (3). Koch's postulates were performed with two representative isolates from each host by artificial inoculation of healthy, green pepper and ripe tomato fruits from the respective cultivars. Fruits were wound inoculated with a sterile scalpel, and small agar plugs (3 to 4 mm) containing 7-day-old sporulating cultures were placed on each wound (five fruits per isolate), or by pipette tip-pricking and pipetting a 5-μl droplet of a conidial suspension (5 × 10 conidia ml) on each wound. The same number of wounded, noninoculated fruits was used as a control. Fruits were maintained in a humidity chamber at 22 to 25°C, and 4 days later, sunken necrotic zones were observed around the wounds of inoculated fruit, whereas control fruits remained symptomless. The pathogen was subsequently reisolated from the inoculated diseased tissues but not from the control fruits. Species-specific PCR (using primer pair CaInt2/ITS4) (2,4) of genomic DNA from three representative isolates (two from pepper and one from tomato) resulted in an amplification product of 490 bp, specific for C. acutatum, further confirming the identity of the pathogen. To our knowledge, this is the second report of C. acutatum in Bulgaria (1), and the first occurrence of that agent on tomato and pepper in this country. References: (1) S. G. Bobev et al. Plant Dis. 86:1178, 2002. (2) S. Freeman et al. Phytopathology 91:586, 2001. (3) P. S. Gunnell and W. D. Gubler. Mycologia 84:157, 1992. (4) M. L. Lewis Ivey et al. Plant Dis. 88:1198, 2004.
2005年夏末,在保加利亚普罗夫迪夫地区的两个农场中,人们观察到辣椒(品种为Kurtovska kapia和当地品种Ribka)出现了零星且异常的损害,在一个农场的番茄(品种为Florida 47)果实上也出现了类似情况。在辣椒果实上观察到干燥、圆形、凹陷的区域(10至20毫米),即使在出现含有橙色分生孢子团的黑色分生孢子盘后,这些区域仍保持其自然果皮颜色。最终,病斑变为褐色,相互融合,果实干枯在植株上。番茄果实也出现了类似症状,但变色不太明显,分生孢子盘也较少。该病原菌很容易从两种寄主植物上分离出来,在马铃薯葡萄糖琼脂培养基上,它形成白色至灰色菌落,平板背面带有鲑鱼橙色色素沉着。形成的分生孢子无色透明,呈梭形,无隔膜,辣椒分离株的分生孢子大小为13.3至17.4×3.5至5.5微米,番茄分离株的分生孢子大小为11.6至15.5×4.1至5.0微米。分离株的培养和形态特征与尖孢炭疽菌(Colletotrichum acutatum)的描述相似(3)。通过对来自每个寄主的两个代表性分离株进行柯赫氏法则验证,将其人工接种到各自品种的健康青椒和成熟番茄果实上。用无菌手术刀对果实进行创伤接种,然后在每个伤口上放置含有7天龄产孢培养物的小琼脂块(3至4毫米)(每个分离株接种5个果实),或者用移液器尖端刺伤果实,并在每个伤口上滴加5微升分生孢子悬浮液(5×10个分生孢子/毫升)。同样数量的创伤但未接种的果实用作对照。将果实置于湿度培养箱中,温度保持在22至25°C,4天后,在接种果实的伤口周围观察到凹陷的坏死区域,而对照果实仍无症状。随后从接种的患病组织中重新分离出病原菌,但对照果实中未分离到。对三个代表性分离株(两个来自辣椒,一个来自番茄)的基因组DNA进行种特异性PCR(使用引物对CaInt2/ITS4)(2,4),得到了一条490 bp的扩增产物,该产物对尖孢炭疽菌具有特异性,进一步证实了病原菌的身份。据我们所知,这是保加利亚关于尖孢炭疽菌的第二篇报道(1),也是该国首次报道该病原菌侵染番茄和辣椒。参考文献:(1)S. G. Bobev等人,《植物病害》,86:1178,2002年。(2)S. Freeman等人,《植物病理学》,91:586,2001年。(3)P. S. Gunnell和W. D. Gubler,《真菌学》,84:157,1992年。(4)M. L. Lewis Ivey等人,《植物病害》,88:1198,2004年。