González V, Tuset J J, Hinarejos R
Department of Plant Protection and Biotechnology, Instituto Valenciano de Investigaciones Agrarias (IVIA). Moncada, Valencia, Spain.
Plant Dis. 2006 Nov;90(11):1462. doi: 10.1094/PD-90-1462A.
During 2004, the natural juniper forests of the Mediterranean mountain ranges of Castellón and Valencia provinces (Comunidad Valenciana, eastern Spain) were severely affected by wood decay. A 2-year drought was registered in the area, and analyzed soil samples had high rates of conductivity (as much as 1,400 μS/cm) in the vicinity of diseased stands. Disease patches occurred within a 2-ha area (with a density of 5 to 30 stands per hectare), and tree damage varied in these symptomatic stands ranging from having chlorotic foliage (10 to 20% of the total surface) to complete defoliation and death of entire stands. After removing dead trees, a white laminated rot was observed located at their bases, specifically on the collar of the roots and the branches in contact with the substrate. In addition to these symptoms, annual, poroid, resupinate, ferruginous-colored basidiocarps were found on 90% of the infected or dead trees in each stand surveyed. These basidiocarps were characterized by having a dimitic hyphal system without clamp-connections, hymenial and hyphal thick-walled setae. Basidiospores were thin-walled, smooth, oblong, and ranged in size from 5 to 7 × 3 to 3.5 μm (4 × 4 μm, extreme measures). On the basis of these features, fungal fruitbodies were preliminary characterized as belonging to Phellinus ferruginosus (Schrad.: Fr.) Pat. (= Fuscoporia ferruginosa (Schrad.) Murrill.). Besides morphological identifications, isolations from infected wood were obtained. Infected branches of Juniperus communis were transversely sectioned and fragmented in small chips (0.5 × 0.5 cm ) that were surface sterilized and placed on potato dextrose agar (PDA) supplemented with 0.5 mg/ml of streptomycin sulfate. Emerging colonies were then transferred to PDA and malt agar plates and incubated at 26°C for 15 to 20 days with a 12-h photoperiod. Cultures also identified as P. ferruginosus had submerged hyphae with swellings, hyphal setae, and ochraceous, tawny-to-honey yellow colonies (3). To confirm pathogenicity, healthy 2-year-old J. communis seedlings (40 replicates) were artificially inoculated with P. ferruginosus (isolate PHE-05-S30). Inoculum was grown on millet grains colonized by the fungus under sterile conditions and subsequently dehydrated. Two grams of the millet grain were applied at the base of the stems and 2 grams were mixed with potting substrate. Pots were covered with plastic bags for 5 days and incubated at room temperature for an additional 25 days. After this period, P. ferruginosus was reisolated (90% recovery) from rotten pieces of wood placed on petri dishes containing PDA, thus fulfilling Koch's postulates. In Europe, this fungus was previously reported to occur on living J. communis in southern Italy (1) and could be distinguished from other closely related Phellinus spp. (2). To our knowledge, this is the first report of white laminated rot on Juniperus spp. caused by P. ferruginosus in Spain. References: (1) A. Bernicchia. Mycotaxon 75:241, 2000. (2) L. Ryvarden and R. L. Gilbertson. European Polypores. Part 2: Meripilus-Tyromyces. Synopsis Fungorum. Vol. 7. Fungiflora, Oslo, Norway, 1994. (3) J. A. Stalpers. Stud. Mycol. 16:1, 1978.
2004年期间,西班牙东部巴伦西亚自治区(Comunidad Valenciana)卡斯特利翁省和巴伦西亚省地中海山脉的天然杜松林受到木材腐朽的严重影响。该地区出现了持续两年的干旱,对土壤样本的分析显示,患病林分附近土壤的电导率很高(高达1400 μS/cm)。病害斑块出现在一个2公顷的区域内(每公顷有5至30株林分),这些有症状的林分中树木的损害程度各不相同,从叶片褪绿(占总表面积的10%至20%)到整片林分完全落叶和死亡。清除死树后,在树基部观察到白色层状腐朽,特别是在根颈和与基质接触的枝条处。除了这些症状外,在每个调查林分中,90%的受感染或死亡树木上发现了一年生、多孔、平伏、铁锈色的担子果。这些担子果的特征是具有双型菌丝系统,无锁状联合,有子实层和菌丝厚壁刚毛。担孢子薄壁、光滑、长圆形,大小范围为5至7×3至3.5μm(极端尺寸为4×4μm)。基于这些特征,真菌子实体初步鉴定为属于铁锈色木层孔菌(Phellinus ferruginosus (Schrad.: Fr.) Pat.)(= 铁锈色褐层孔菌(Fuscoporia ferruginosa (Schrad.) Murrill.))。除了形态学鉴定外,还从受感染的木材中进行了分离培养。将杜松(Juniperus communis)的受感染枝条横向切片并切成小碎片(0.5×0.5厘米),进行表面消毒后置于添加了0.5毫克/毫升硫酸链霉素的马铃薯葡萄糖琼脂(PDA)上。然后将长出的菌落转移到PDA和麦芽琼脂平板上,在26°C下、12小时光周期条件下培养15至20天。鉴定为铁锈色木层孔菌的培养物具有带膨大的浸没菌丝、菌丝刚毛以及淡褐色至黄褐色的菌落(3)。为了确认致病性,用铁锈色木层孔菌(分离株PHE - 05 - S30)人工接种健康的2年生杜松幼苗(40个重复)。接种物在无菌条件下在被该真菌定殖的小米粒上生长,随后进行脱水处理。将2克小米粒施用于茎基部,另外2克与盆栽基质混合。花盆用塑料袋覆盖5天,然后在室温下再培养25天。在此之后,从放置在含有PDA的培养皿中的腐烂木块上再次分离出铁锈色木层孔菌(回收率为90%),从而满足了柯赫氏法则。在欧洲,此前曾报道该真菌在意大利南部的活杜松上出现(1),并且可以与其他近缘的木层孔菌属物种区分开来(2)。据我们所知,这是西班牙首次关于铁锈色木层孔菌引起杜松属植物白色层状腐朽的报道。参考文献:(1)A. Bernicchia. Mycotaxon 75:241, 2000.(2)L. Ryvarden和R. L. Gilbertson. European Polypores. Part 2: Meripilus - Tyromyces. Synopsis Fungorum. Vol. 7. Fungiflora, Oslo, Norway, 1994.(3)J. A. Stalpers. Stud. Mycol. 16:1, 1978.