Santori A, Vitale S, Luongo L, Belisario A
Centro di Ricerca per la Patologia Vegetale (CRA-PAV), Via C. G. Bertero 22, 00156 Rome, Italy.
Plant Dis. 2010 Apr;94(4):484. doi: 10.1094/PDIS-94-4-0484A.
Hazelnut (Corylus avellana) is a traditionally cultivated nut species in Italy. Italy is the second largest producer of hazelnut in the world after Turkey. In early summer of 2000, a severe fruit drop (up to 60%) was observed in several hazelnut orchards located in the Latium Region in central Italy. The severity of yield losses led to investigating the etiology of the disease subsequently named nut gray necrosis (NGN) based on symptoms observed on the affected fruit. Symptoms consisted of a brown grayish necrotic spot/patch on the nut shell and bracts, sometimes involving the petiole (1). Isolations of the potential pathogen were from tissue that was sampled starting from bloom of female flowers to fully ripened fruit. Isolations from symptomatic tissue consisted of placing onto potato dextrose agar (PDA) small tissue fragments (approximately 3 mm) cut from the margin of lesions, while asymptomatic material (entire flowers or young fruit) was sectioned into small pieces. All the material was previously surface disinfected with 1% NaOCl. Slow-growing, dark grayish olive colonies were obtained consistently within 14 days of incubation at 20 to 22°C from symptomatic and asymptomatic material. Sporodochia were rarely produced on PDA, but never on carnation leaf agar. Dark grayish olive colonies were assigned to a Fusarium sp. Detached hazelnut fruit exposed to 20 μl of a mycelial suspension (10 CFU/ml) and incubated in a moist chamber at room temperature for 10 days produced orange sporodochia bearing 3 to 5 septate macroconidia (35 × 4 μm). On the basis of morphology, the fungus was identified as Fusarium lateritium Nees. (3,4). The identity was confirmed by internal transcribed spacer rDNA sequence comparison with BBA65248 (GenBank Accession No. AF310980). The sequences of two isolates, ISPaVe1874 and ISPaVe1976, were deposited in GenBank (Accession Nos. FN547420 and FN547445, respectively). Pathogenicity tests were performed in planta by inoculating, with the aforementioned isolates, young to fully formed fruit (approximately 24 mm in diameter) either with a drop of mycelial (10 CFU/ml) or conidial (10 conidia/ml) suspension. Drops were placed between the nut and leafy involucre. Controls were treated with sterile water only. Within 2 weeks after inoculation, a grayish necrosis developed on all the inoculated fruit and was similar to symptoms originally observed in the field. No differences were observed between the two methods of inoculation. On full-sized fruit, lesions extended from the shell to inner tissues. The pathogen was consistently reisolated from lesions. Controls showed no symptoms. To our knowledge, this is the first report of F. lateritium as the causal agent of nut gray necrosis on hazelnut in Italy, and this pathogen has never been reported as an agent of necrosis and drop of hazelnut fruit, but it was previously reported as an agent of twig cankers (2). References: (1) A. Belisario et al. Inf. Agrario 59:71, 2003. (2) A. Belisario et al. Plant Dis. 89:106, 2005. (3) J. F. Leslie et al. The Fusarium Laboratory Manual. Blackwell Publishing Press Ltd, Oxford, UK, 2006. (4) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University Press, 1983.
榛子(欧洲榛)是意大利传统种植的坚果品种。意大利是仅次于土耳其的世界第二大榛子生产国。2000年夏初,在意大利中部拉齐奥地区的几个榛果园中观察到严重的落果现象(高达60%)。产量损失的严重程度促使人们随后基于受影响果实上观察到的症状对这种疾病的病因进行调查,该疾病随后被命名为坚果灰坏死病(NGN)。症状包括坚果壳和苞片上出现棕灰色坏死斑/块,有时涉及叶柄(1)。潜在病原菌的分离是从雌花开花到果实完全成熟期间采集的组织进行的。从有症状组织中分离时,是将从病斑边缘切下的小组织碎片(约3毫米)接种到马铃薯葡萄糖琼脂(PDA)上,而无症状材料(整朵花或幼果)则切成小块。所有材料事先都用1%次氯酸钠进行了表面消毒。在20至22°C下培养14天内,从有症状和无症状材料中均持续获得生长缓慢、深灰橄榄色的菌落。在PDA上很少产生分生孢子盘,但在香石竹叶琼脂上从未产生。深灰橄榄色菌落被鉴定为镰刀菌属。将离体榛子果实暴露于20微升菌丝悬浮液(10 CFU/ml)中,并在室温下的保湿箱中培养10天,产生了带有3至5个分隔的大型分生孢子(35×4微米)的橙色分生孢子盘。根据形态学,该真菌被鉴定为砖红镰刀菌Nees。(3,4)。通过与BBA65248(GenBank登录号AF310980)进行内部转录间隔区rDNA序列比较,确认了其身份。两个分离株ISPaVe1874和ISPaVe1976的序列已存入GenBank(登录号分别为FN547420和FN547445)。通过用上述分离株接种直径约24毫米的幼果至成熟果实,对果实进行致病性测试,接种物为一滴菌丝(10 CFU/ml)或分生孢子(10个分生孢子/ml)悬浮液。液滴置于坚果与叶状总苞之间。对照组仅用无菌水处理。接种后2周内,所有接种果实均出现灰坏死,与最初在田间观察到的症状相似。两种接种方法之间未观察到差异。在成熟果实上,病斑从外壳扩展到内部组织。病原菌始终能从病斑中重新分离出来。对照组未出现症状。据我们所知,这是关于砖红镰刀菌作为意大利榛子坚果灰坏死病因的首次报道,该病原菌从未被报道为榛子果实坏死和落果的病原菌,但此前曾被报道为嫩枝溃疡病的病原菌(2)。参考文献:(1)A. Belisario等人,《农业信息》59:71,2003年。(2)A. Belisario等人,《植物病害》89:106,2005年。(3)J. F. Leslie等人,《镰刀菌实验室手册》。英国牛津布莱克韦尔出版有限公司,2006年。(4)P. E. Nelson等人,《镰刀菌物种:鉴定图谱手册》。宾夕法尼亚州立大学出版社,1983年。