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意大利首次报道立枯丝核菌AG-1-IA引起的毛地黄叶枯病

First Report of Leaf Blight on Foxglove (Digitalis purpurea) Caused by Rhizoctonia solani AG-1-IA in Italy.

作者信息

Garibaldi A, Gilardi G, Bertetti D, Gullino M L

机构信息

Center of Competence AGROINNOVA, University of Torino, Via Leonardo da Vinci, 44, 10095 Grugliasco, Italy.

出版信息

Plant Dis. 2009 Mar;93(3):318. doi: 10.1094/PDIS-93-3-0318C.

Abstract

Digitalis purpurea (Scrophulariaceae), foxglove, is used in flower gardens. In the spring of 2008, leaf blight was observed in a nursery near Biella (northern Italy) on 30% of potted 30-day-old plants grown in a peat substrate at temperatures from 20 to 25°C and relative humidity at 75 to 80%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the blade-petiole junction and later along the leaf margins. Lesions expanded for several days along the midvein until the entire leaf was affected. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily recovered, then transferred and maintained in pure culture (4). The isolates of R. solani obtained from affected plants successfully anastomosed with tester isolate AG 1 (ATCC 58946). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates AG 2, 3, 4, 6, 7, 11, and AG BI and anastomosis was not observed. Ten-day-old colonies grown on PDA appeared light brown, rather compact, and radial. Numerous sclerotia of uniform size (0.5 to 3 mm in diameter) and sometimes joined laterally were formed. Descriptions of mycelium and sclerotia were typical for subgroup IA Type 2 (4). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 724-bp fragment showed a 99% homology with the sequence of R. solani (GenBank Accession No. EU591800). The nucleotide sequence has been assigned GenBank Accession No. FJ467490. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Plants of 30-day-old D. purpurea were grown in 10-liter containers (6 plants per container) in a steam disinfested peat/clay/perlite (70:20:10) substrate. Disks of PDA cultures were placed on leaves (1 cm of mycelium per plant). Plants inoculated with PDA alone served as control treatments. Three replicates were used. Plants were maintained in a growth chamber at 24 ± 1°C with 12 h light/dark. First symptoms developed 12 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was repeated twice. R. solani was isolated from a small percentage of infected seeds of D. purpurea in India (3). This is, to our knowledge, the first report of leaf blight of D. purpurea caused by R. solani in Italy as well as in Europe. The spread of R. solani in nurseries might cause a decrease in trade. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions in: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, the Netherlands, 1996. (3) K. K. Janardhanan and D. Ganguly. Indian Phytopathol. 16:379, 1963. (4) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

摘要

毛地黄(玄参科),即洋地黄,常用于花园。2008年春季,在意大利北部比耶拉附近的一家苗圃中,观察到30%的盆栽30日龄植株出现叶枯病,这些植株种植在泥炭基质中,温度为20至25°C,相对湿度为75至80%。叶片在土壤线以上、叶片与叶柄交界处刚出现半圆形、水渍状病斑,随后沿叶缘扩展。病斑沿叶脉扩展数天,直至整片叶子都受到影响。枯萎的叶子变成褐色,枯萎,附着在茎上,并与周围的叶子交织在一起。将患病组织在1%次氯酸钠中消毒10秒,用无菌水冲洗,然后接种在添加了100毫克/升硫酸链霉素的马铃薯葡萄糖琼脂(PDA)上。始终能很容易地分离出具有立枯丝核菌形态特征的真菌,然后将其转移并保存在纯培养物中(4)。从受影响植株上获得的立枯丝核菌分离物与测试分离物AG 1(ATCC 58946)成功融合。融合点处的菌丝直径减小,融合点明显,观察到相邻细胞死亡。结果与其他关于融合反应的报道一致(2)。还与测试分离物AG 2、3、4、6、7、11和AG BI进行配对,未观察到融合现象。在PDA上生长10天的菌落呈浅褐色,相当致密,呈放射状。形成了许多大小均匀(直径0.5至3毫米)、有时横向相连的菌核。菌丝体和菌核的描述是IA亚组2型的典型特征(4)。使用引物ITS4/ITS6扩增rDNA的内部转录间隔区(ITS)并进行测序。对724个碱基对片段的BLASTn分析(1)显示与立枯丝核菌序列(GenBank登录号EU591800)有99%的同源性。该核苷酸序列已被赋予GenBank登录号FJ467490。为进行致病性测试,将立枯丝核菌接种物通过在PDA上培养病原体10天来制备。30日龄的毛地黄植株种植在10升容器(每个容器6株植物)中,基质为经过蒸汽消毒的泥炭/粘土/珍珠岩(70:20:10)。将PDA培养物圆盘放置在叶片上(每株植物1厘米菌丝体)。仅接种PDA的植株作为对照处理。使用三个重复。将植株置于生长室中,温度为24±1°C,光照/黑暗周期为12小时。人工接种后12天出现最初症状。始终能从受感染的叶片和茎中重新分离出立枯丝核菌。对照植株保持健康。致病性测试重复两次。在印度,从一小部分受感染的毛地黄种子中分离出了立枯丝核菌(3)。据我们所知,这是意大利以及欧洲首次关于立枯丝核菌引起毛地黄叶枯病的报道。立枯丝核菌在苗圃中的传播可能会导致贸易量下降。参考文献:(1)S. F. Altschul等人,《核酸研究》25:3389,1997年。(2)D. E. Carling,《通过菌丝融合反应对立枯丝核菌进行分组》,载于《立枯丝核菌物种:分类学、分子生物学、生态学、病理学和病害控制》,荷兰Kluwer学术出版社,1996年。(3)K. K. Janardhanan和D. Ganguly,《印度植物病理学》16:379,1963年。(4)B. Sneh等人,《立枯丝核菌物种的鉴定》,美国植物病理学会,明尼苏达州圣保罗,1991年。

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