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加拿大艾伯塔省杨树青铜叶病的首次报道及杨树生拟盘多毛孢菌的序列分析

First Report of Bronze Leaf Disease on Poplar in Alberta, Canada and Sequence of Apioplagiostoma populi.

作者信息

Kawchuk L M, Howard R J, Kalischuk M L, Northover P R, Desjardins M, Spencer R C J

机构信息

Agriculture and Agri-Food Canada, Lethbridge, AB, T1J 4B1, Canada.

Alberta Agriculture and Rural Development, Crop Diversification Centre South, Brooks, AB T1R 1E6, Canada.

出版信息

Plant Dis. 2010 Mar;94(3):377. doi: 10.1094/PDIS-94-3-0377A.

Abstract

Poplar (Populus spp.) is an important ornamental, windbreak, and pulp and wood product tree in Alberta and across western Canada because of its rapid growth, architecture, and hardiness. It is also a major component of native tree stands in the parkland area of the Canadian Prairies. Until recently in North America, infections of Apioplagiostoma populi (Cash & A.M. Waterman) Barr have only been documented in central Canada and the eastern and midwestern United States. Symptoms resembling bronze leaf disease (3) were observed in Alberta as early as 2003 and have been seen each subsequent year on an increasing number of Populus × canescens Smith, P. tremula L., and P. tremuloides Michx. trees from urban areas, shelterbelts, and nurseries. Foliar symptoms were observed in 10 to 50% of the tree canopy, and diseased leaves were bronze-colored with green and yellow petioles and veins. Disease symptoms became pronounced in mid-to-late summer with bronze to dark reddish brown leaves, while the petiole and the midrib remained green. Some symptomatic leaves remained attached to diseased trees throughout the fall and winter and continued the infectious disease cycle in the spring. As the disease advanced, A. populi colonized stem and branch tissues causing the leaves to wilt, discolor, and die shortly afterward. Diseased branches often died within the current season. Continued branch dieback resulted in significantly reduced aesthetic and commercial value. Survival of poplar arising from diseased clones was often less than 5 years. Bronze leaf disease symptoms have been reported on several Populus spp., and premature tree mortality represents a serious impediment to the continued use of this tree species (1). Attempts to isolate the causal agent of bronze leaf disease on artificial media have been unsuccessful (4). In the fall of 2008, leaves from symptomatic trees were collected and suspended outdoors in mesh bags to overwinter. Dark brown perithecia (150 to 200 × 100 to 150 μm) emerged the following spring from the lower and upper leaf surfaces. Asci were fusoid clavate, 30 to 40 × 10 to 14 μm with a conspicuous apical ring and contained hyaline two-celled ascospores 10 to 14 × 3 to 6 μm that were ellipsoid clavate with a relatively short basal cell. Nucleic acid was extracted from isolated perithecia and amplified by the polymerase chain reaction and oligonucleotides 5'GCATCGATGAAGAACGCAGC3' and 5'TCCTCCGCTTATTGATATGC3' specific for rDNA internal transcribed spacer (ITS) sequence (2). The cloned amplified sequence of the A. populi rDNA ITS region (GenBank Accession No. GU205341) showed considerable homology (>90% identity) to other Apioplagiostoma spp. In total, 33 independent leaf samples from nine trees exhibiting disease symptoms were positive for A. populi, producing an approximately 300-bp sequence not observed in any of the symptomless samples. Poplar and aspen have been extensively planted in rural and urban landscapes in western Canada over the past 100 years and continued spread of the bronze leaf disease pathogen threatens the viability of the shelterbelt, nursery, and processed wood industries. References: (1) E. K. Cash and A. M. Waterman. Mycologia 49:756, 1957. (2) A. H. Khadhair et al. Can. J. Plant Pathol. 20:55, 1998. (3) P. R. Northover and M. Desjardins. Plant Dis. 87:1538, 2003. (4) J. A. Smith et al. Plant Dis. 86:462, 2002.

摘要

杨树(Populus spp.)因其生长迅速、树形美观且耐寒,在艾伯塔省及加拿大西部是一种重要的观赏树、防风树以及纸浆和木材原料树。它也是加拿大大草原公园地区原生林分的主要组成部分。直到最近在北美,杨叶单囊壳菌(Apioplagiostoma populi (Cash & A.M. Waterman) Barr)感染仅在加拿大中部以及美国东部和中西部有记录。早在2003年,在艾伯塔省就观察到了类似青铜叶病(3)的症状,并且在随后的每一年,城市地区、防护林带和苗圃中越来越多的灰杨(Populus × canescens Smith)、欧洲山杨(P. tremula L.)和颤杨(P. tremuloides Michx.)树上都出现了这种症状。在树冠的10%至50%观察到了叶片症状,患病叶片呈青铜色,叶柄和叶脉为绿色和黄色。病害症状在夏中晚期变得明显,叶片从青铜色变为深红褐色,而叶柄和中脉仍保持绿色。一些有症状的叶片在整个秋冬季节都附着在患病树上,并在春季继续传染病循环。随着病害发展,杨叶单囊壳菌定殖于茎和枝组织,导致叶片不久后枯萎、变色并死亡。患病枝条通常在当季死亡。枝条持续枯死导致美观和商业价值显著降低。由患病无性系产生的杨树存活期通常不到5年。在几种杨树品种上都报告了青铜叶病症状,树木过早死亡对该树种的持续利用构成了严重障碍(1)。在人工培养基上分离青铜叶病病原体的尝试均未成功(4)。2008年秋季,采集了有症状树木的叶片,装在网袋中挂在户外越冬。次年春天,在叶片的上下表面出现了深褐色的子囊壳(150至200×100至150μm)。子囊呈梭形棒状,30至40×10至14μm,有明显的顶端环,内含透明的双细胞子囊孢子,10至14×3至6μm,呈椭圆形棒状,基部细胞相对较短。从分离的子囊壳中提取核酸,通过聚合酶链反应和针对核糖体DNA内转录间隔区(ITS)序列的寡核苷酸5'GCATCGATGAAGAACGCAGC3'和5'TCCTCCGCTTATTGATATGC3'进行扩增(2)。杨叶单囊壳菌核糖体DNA ITS区域的克隆扩增序列(GenBank登录号GU205341)与其他杨叶单囊壳菌属物种显示出相当高的同源性(>90%同一性)。总共,来自9棵表现出病害症状树木的33个独立叶片样本对杨叶单囊壳菌呈阳性,产生了一个在任何无症状样本中都未观察到的约300bp序列。在过去100年里,杨树和山杨在加拿大西部的农村和城市景观中广泛种植,青铜叶病病原体的持续传播威胁着防护林带、苗圃和加工木材行业的生存能力。参考文献:(1)E. K. Cash和A. M. Waterman。《真菌学》49:756,1957。(2)A. H. Khadhair等人。《加拿大植物病理学杂志》20:55,1998。(3)P. R. Northover和M. Desjardins。《植物病害》87:1538,2003。(4)J. A. Smith等人。《植物病害》86:462,2002。

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