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茄病镰刀菌引起阿根廷东南部橄榄根腐病的首次报道

First Report of Fusarium solani Causing Root Rot of Olive in Southeastern Argentina.

作者信息

Perez B A, Farinon O M, Berretta M F

机构信息

INTA-CICVyA-IMYZA, Nicolas Repetto y De Los Reseros (1686) Hurlingham, Buenos Aires, Argentina.

出版信息

Plant Dis. 2011 Nov;95(11):1476. doi: 10.1094/PDIS-02-11-0095.

Abstract

In Argentina, olive (Olea europaea L.) is cultivated in the mountainous, warm, arid northwest (Andes range), where Fusarium solani (blue sporodochia) is frequently found to be causing death of nursery and young field plants (1). Recently, olive orchards were established more than 1,600 km to the southeast (Pampas) in a plain with a temperate and humid climate and in the arid Patagonia, both influenced by the Atlantic Ocean. This area includes part of Buenos Aires and Rio Negro provinces. In March 2008, 10-year-old 'Barnea' olive trees with high incidence of root rot, dried leaves, dead branches, and dead plants were observed in the Coronel Dorrego District of Buenos Aires Province, where oat, barley or other cereals are planted between rows of olive trees. Planting material originated from olive nurseries located in Mendoza Province, 1,200 km from Coronel Dorrego. Diseased roots were disinfected in 2% NaOCl and 70% ethanol, cut into small pieces, plated onto rose bengal-glycerin-urea medium, and incubated at 20°C with a 12-h photopheriod. A fungus was purified through successive transfers of hyphal tips from the margin of a sparsely growing colony onto 2% water agar (2). Colonies grown on Spezieller Nährstoffarmer agar (3) and carnation leaf-piece agar were used for morphological identification, and those on grown on potato dextrose agar were used for evaluation of pigmentation and colony growth rate. Sporodochium color, cream, was typical of F. solani (Mart.) Sacc. This isolate was deposited in the IMYZA Microbial Collection as INTA-IMC 73. Mycelium was cultured in liquid Czapek-Dox medium supplemented with sucrose, peptone, yeast extract, sodium nitrate, and vitamins for 4 days and fungal DNA was obtained with a DNA extraction kit. Primers ITS1 and ITS4 were used to amplify the internal transcribed spacer (ITS) region of ribosomal genes. The purified PCR product was sequenced and the DNA sequence compared with GenBank records. The sequence shared 100% identity with 27 entries for F. solani and 97% identity with F. solani obtained from olive in Nepal (4), corresponding to EU912432 and EU912433. The nucleotide sequence was registered in GenBank as JF299258. Pathogenicity was confirmed on 'Manzanilla' plants at the eight-leaf stage. Pieces of water agar with mycelium were applied to small wounds at the stem base and on roots of 10 plants and were covered with cotton soaked in sterile distilled water. Plants were incubated at 20°C and a 14-h photoperiod. On control plants, water agar pieces without mycelium were applied to the wounds. After 33 days, inoculated plants showed dark brown lesions (average length 1.4 cm) and leaf chlorosis. Two plants showed wilting with leaves remaining attached to branches. F. solani was reisolated from roots and stem bases of inoculated plants. Controls remained asymptomatic. To our knowledge, this is the first report of F. solani occurring on olive in the temperate part of the Pampas of Argentina where cereals, which are susceptible to Fusarium species, are grown with olive trees. Sporodochium color (cream) of these isolates differed from the blue color of previously reported isolates of F. solani on olive in northwestern Argentina (1). References: (1) S. Babbitt et al. Plant Dis. 86:326, 2002. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) H. I. Nirenberg. Releases Fed. Biol. Res. Center Agric. For. (Berlin-Dahlem) 169:1, 1976. (4) A. M. Vettraino et al. Plant Dis. 23:200, 2009.

摘要

在阿根廷,油橄榄(油橄榄属)种植于多山、温暖且干旱的西北部地区(安第斯山脉),在那里,经常发现茄病镰刀菌(蓝色分生孢子座)致使苗圃和田间幼株死亡(1)。最近,在东南部1600多公里处的平原(潘帕斯草原)以及受大西洋影响的干旱巴塔哥尼亚地区建立了油橄榄果园。该地区包括布宜诺斯艾利斯省和内乌肯省的部分地区。2008年3月,在布宜诺斯艾利斯省科罗内尔多雷戈区观察到10年生的‘巴尔内亚’油橄榄树,根腐病发病率高,树叶干枯,树枝死亡,植株死亡,该地区在油橄榄树行间种植燕麦、大麦或其他谷物。种植材料来自距离科罗内尔多雷戈1200公里的门多萨省的油橄榄苗圃。将患病根系在2%次氯酸钠和70%乙醇中消毒,切成小块,接种到孟加拉玫瑰 - 甘油 - 尿素培养基上,并在20°C、12小时光照周期下培养。通过将菌丝尖端从稀疏生长菌落的边缘连续转移到2%水琼脂上对一种真菌进行纯化(2)。在特殊营养贫乏琼脂(3)和香石竹叶片琼脂上生长的菌落用于形态学鉴定,在马铃薯葡萄糖琼脂上生长的菌落用于色素沉着和菌落生长速率评估。分生孢子座颜色为奶油色,是茄病镰刀菌(Mart.)Sacc.的典型特征。该分离株作为INTA - IMC 73保藏于IMYZA微生物保藏中心。将菌丝体在添加蔗糖、蛋白胨、酵母提取物、硝酸钠和维生素的液体察氏培养基中培养4天,并用DNA提取试剂盒获得真菌DNA。使用引物ITS1和ITS4扩增核糖体基因的内部转录间隔区(ITS)。对纯化的PCR产物进行测序,并将DNA序列与GenBank记录进行比较。该序列与茄病镰刀菌的27个条目具有100%的同一性,与从尼泊尔油橄榄中获得的茄病镰刀菌具有97%的同一性(4),对应于EU912432和EU912433。核苷酸序列在GenBank中注册为JF299258。在八叶期的‘曼萨尼拉’植株上证实了致病性。将带有菌丝体的水琼脂片应用于10株植物茎基部和根部的小伤口上,并用无菌蒸馏水浸泡的棉花覆盖。将植株在20°C、14小时光照周期下培养。在对照植株上,将没有菌丝体的水琼脂片应用于伤口。33天后,接种植株出现深褐色病斑(平均长度1.4厘米)和叶片萎黄。两株植株出现萎蔫,叶片仍附着在枝条上。从接种植株的根和茎基部重新分离出茄病镰刀菌。对照植株无症状。据我们所知,这是茄病镰刀菌在阿根廷潘帕斯草原温带地区油橄榄上发生情况的首次报道,该地区种植易感染镰刀菌属的谷物和油橄榄树。这些分离株的分生孢子座颜色(奶油色)与先前报道的阿根廷西北部油橄榄上的茄病镰刀菌分离株的蓝色不同(1)。参考文献:(1)S. Babbitt等人,《植物病害》86:326,2002年。(2)J. F. Leslie和B. A. Summerell,《镰刀菌实验室手册》,Blackwell Publishing,艾姆斯,爱荷华州,2006年。(3)H. I. Nirenberg,《联邦生物研究中心农业与林业通报》(柏林 - 达勒姆)169:1,1976年。(4)A. M. Vettraino等人,《植物病害》23:200,2009年。

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