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烟草疫霉引起的豆瓣绿根腐病和茎腐病在中国的首次报道

First Report of Root and Stem Rot Caused by Phytophthora nicotianae on Peperomia tetraphylla in China.

作者信息

Zeng D X, Wu X L, Li Y H

机构信息

School of Applied Chemistry and Biological Technology, Shenzhen Polytechnic, Shenzhen 518055, China.

出版信息

Plant Dis. 2010 Sep;94(9):1171. doi: 10.1094/PDIS-94-9-1171C.

Abstract

Peperomia tetraphylla, an evergreen herb, is becoming increasingly popular as a potted ornamental plant in southern China. In the summer of 2008, in some commercial flower nurseries in Shenzhen, Guangdong Province, P. tetraphylla showed extensive black stem and root rot, with leaves dropping from the rotten stem. Small pieces (approximately 3 mm) of stems and leaves were excised from the margins of the black lesions, surface disinfected for 30 s to 1 min in 0.1% HgCl, plated onto potato dextrose agar (PDA), and incubated at 25°C in the dark. All the plated samples yielded Phytophthora, and microscopic examination of pure cultures grown on PDA plates showed arachnoid colonies with abundant aerial mycelium, chlamydospores, and a few sporangia. Numerous sporangia were formed in sterile soil extract. Sporangia were ovoid or obpyriform, noncaducous, with prominent solitary papillae, and measured 31 to 52 μm (average 38 μm) × 21 to 34 μm (average 27 μm). Chlamydospores were spherical and 21 to 34 μm in diameter (average 28 μm). The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using primers ITS/ITS and sequenced (2). The ITS sequence, when submitted for a BLAST search in the NCBI database, showed 100% homology with the sequences of two reference isolates of Phytophthora nicotianae (Accession Nos. AY833526 and EU433396) and the consensus ITS sequence was deposited in the NCBI as Accession No. GQ499373. The isolate was identified as Phytophthora nicotianae on the basis of morphological and molecular characteristics (1). Pathogenicity of the isolate was confirmed by inoculating 1-year-old plants of P. tetraphylla growing in pots. The isolate was grown for 7 days on PDA plates and mycelial plugs, 5 mm in diameter and taken from the advancing margins of the colonies, were buried approximately 1 cm deep near the base of the stem in such a way that the mycelium on the plugs was in contact with the surface of the stem, which had been wiped earlier with 70% ethanol and gently wounded with a needle. Plants treated the same way but inoculated with sterile PDA plugs served as control plants. Three plants in each pot were inoculated and there were five replications each for the treatment and the control. All plants were kept in a greenhouse at 22 to 32°C. After 6 to 7 days, the inoculated plants showed black lesions around the mycelial plugs; symptoms of root and stem rot developed rapidly thereafter and the plants collapsed within 2 weeks. All symptoms on the inoculated plants were identical to those observed in naturally diseased plants, whereas the control plants remained healthy. The same fungus was consistently reisolated from the inoculated plants. To our knowledge, this is the first report of Phytophthora nicotianae on P. tetraphylla in China. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) J. B. Ristaino et al. Appl. Environ. Microbiol. 64:948, 1998.

摘要

豆瓣绿是一种常绿草本植物,在中国南方作为盆栽观赏植物越来越受欢迎。2008年夏天,在广东省深圳市的一些商业花卉苗圃中,豆瓣绿出现了广泛的茎部和根部黑腐病,叶片从腐烂的茎上掉落。从黑色病斑边缘切取小块(约3毫米)茎和叶,在0.1%升汞中表面消毒30秒至1分钟,接种到马铃薯葡萄糖琼脂(PDA)平板上,于25°C黑暗条件下培养。所有接种的样品都分离出了疫霉属真菌,对在PDA平板上生长的纯培养物进行显微镜检查,发现有蛛网状菌落,有丰富的气生菌丝、厚垣孢子和一些孢子囊。在无菌土壤浸出液中形成了大量孢子囊。孢子囊呈卵形或倒梨形,不脱落,有明显的单个乳突,大小为31至52微米(平均38微米)×21至34微米(平均27微米)。厚垣孢子呈球形,直径为21至34微米(平均28微米)。使用引物ITS/ITS对单个分离物的核糖体DNA内部转录间隔区(ITS)进行扩增并测序(2)。将ITS序列提交到NCBI数据库进行BLAST搜索时,发现其与烟草疫霉两个参考分离物的序列(登录号AY833526和EU433396)有100%的同源性,其ITS序列的一致序列作为登录号GQ499373保存在NCBI中。根据形态学和分子特征,该分离物被鉴定为烟草疫霉(1)。通过对接种在花盆中生长的1年生豆瓣绿植株进行接种,证实了该分离物的致病性。将该分离物在PDA平板上培养7天,从菌落生长前沿切取直径5毫米的菌丝块,埋在茎基部附近约1厘米深处,使菌丝块上的菌丝与茎表面接触,茎表面事先用70%乙醇擦拭并轻轻用针刺伤。以同样方式处理但接种无菌PDA菌丝块的植株作为对照植株。每盆接种3株,处理和对照各有5次重复。所有植株都保存在22至32°C的温室中。6至7天后,接种的植株在菌丝块周围出现黑色病斑;此后根腐和茎腐症状迅速发展,植株在2周内枯萎。接种植株上的所有症状与自然发病植株上观察到的症状相同,而对照植株保持健康。从接种植株上持续重新分离出相同的真菌。据我们所知,这是中国关于烟草疫霉侵染豆瓣绿的首次报道。参考文献:(1)D.C.欧文和O.K.里贝罗。《世界疫霉病》。美国植物病理学会,明尼苏达州圣保罗,1996年。(2)J.B.里斯泰诺等人。《应用与环境微生物学》64:948,1998年。

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