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意大利常见鼠尾草(Salvia officinalis L.)上的隐地疫霉。

Phytophthora cryptoea on Common Sage (Salvia officinalis L.) in Italy.

作者信息

Garibaldi A, Bertetti D, Pensa P, Ortu G, Gullino M L

机构信息

Centre of Competence for the Innovation in the Agro-Environmental sector, AGROINNOVA, University of Torino, Grugliasco, Italy.

Centre of Competence for the Innovation in the Agro-Environmental sector, AGROINNOVA, University of Torino, Grugliasco, Italy, and Dipartimento di Scienze Agrarie, Forestali e Alimentari, University of Torino, Grugliasco, Italy.

出版信息

Plant Dis. 2015 Jan;99(1):161. doi: 10.1094/PDIS-09-14-0902-PDN.

Abstract

During the spring of 2013, many plants of common sage (Salvia officinalis L.), grown as potted plants in a commercial farm at Albenga (northern Italy) showed extensive symptoms of foliar wilt and root rot. The first symptoms developed with temperatures ranging between 8 and 26.5°C, average 17°C, and consisted of leaf chlorosis, wilting, and collapse. Severe root and crown rot were also observed, leading to sudden collapse of approximately 60% of the 6,000 plants within 60 days from transplant. Symptomatic tissues from the root and collar of infected plants were surface disinfested for 1 min in a 1% NaOCl solution, rinsed for 5 min in water, and placed on a selective medium for oomycetes (3). A Phytophthora-like organism (1) was consistently isolated and was transferred to carrot agar. Mycelial disks of the isolate DB13GIU02 were floated in petri plates containing soil extract (1), under continuous fluorescent light at room temperature. Hyphal swelling was abundant in such aqueous medium, measuring 6.4 to 20.1 (13.1 average) μm. Sporangia were obpyriform, persistent, and nonpapillate, measuring 25.3 to 55.1 × 17.9 to 37.1 (average 42.8 to 27.9) μm. Oospores and chlamydospores were absent. The same isolate was tested with two isolates of P. cryptogea from Quercus ilex (PH050, mating type A1) and from Pistacia lentiscus (PH017, mating type A2) on carrot agar, at 23 ± 1°C in the dark. Only the paring of DB13GIU02 with PH017 was successful and produced oogonia with diameter of 28.3 to 34.6 (average 31.7) μm, oospores with diameter of 28.0 to 32.2 (average 29.2) μm, and anphigynous antheridia of 10.5 to 15.1 × 11.6 to 15.1 (average 13.5 × 13.3) μm. DNA of the three isolates was extracted by using the Nucleospin Plant kit (Macherey Nagel). PCR of DNA amplified with primers Cryp 1 and Cryp 2 (4) from all P. cryptogea isolates produced a specific amplicon. The internal transcribed spacer (ITS) region of rDNA of the isolate DB13GIU02 was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis of the 845-bp segment (GenBank Accession No. KM458193) showed a 99% homology with the sequence of P. cryptogea GU111631. Pathogenicity tests were performed on healthy common sage 60-day-old plants by using one strain of P. cryptogea grown on a mixture of 2:1 wheat/hemp kernels. Infested kernels (10 g/liter of substrate) were mixed into a steam-disinfested substrate based on sphagnum peat/pomix/pine bark/clay (50:20:20:10 v/v). Control plants were treated with uninoculated wheat/hemp kernels mixed into the steam-disinfested soil. The trial was repeated once. Fifteen plants per treatment were used. All plants were kept in a growth chamber at 20 ± 1°C. Inoculated plants became chlorotic 7 days after inoculation, and root and crown rot developed 15 days after inoculation. P. cryptogea was consistently reisolated from inoculated plants. No colonies were isolated on the selective medium from control plants that remained symptomless. P. cryptogea has been reported on S. officinalis in the United States (2), while in Italy the same pathogen has been observed on S. leucantha. This is the first report of P. cryptogea on S. officinalis in Italy. The economic importance of the disease can increase due to the expanding use of this plant both as an aromatic for culinary purposes and for landscaping. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. APS Press, St. Paul, MN, 1996. (2) S. T. Koike et al. Plant Dis. 81:959, 1997. (3) H. Masago et al. Phytopathology 67:25, 1977. (4) D. Minerdi et al. Eur. J. Plant Pathol. 122:227, 2008.

摘要

2013年春季,在意大利北部阿尔本加的一个商业农场作为盆栽植物种植的许多鼠尾草(Salvia officinalis L.)植株出现了广泛的叶部枯萎和根腐症状。最初症状出现在温度为8至26.5°C(平均17°C)时,表现为叶片黄化、萎蔫和枯萎。还观察到严重的根腐和冠腐,导致6000株植物中约60%在移栽后60天内突然枯萎。将受感染植株根和颈部的症状组织在1%次氯酸钠溶液中表面消毒1分钟,在水中冲洗5分钟,然后置于卵菌选择性培养基上(3)。持续分离出一种类似疫霉的生物体(1),并将其转移到胡萝卜琼脂上。分离株DB13GIU02的菌丝圆盘漂浮在含有土壤提取物的培养皿中(1),在室温下持续荧光灯下培养。在这种水培养基中菌丝大量肿胀,大小为6.4至20.1(平均13.1)μm。孢子囊倒梨形、持久且无乳突,大小为25.3至55.1×17.9至37.1(平均42.8至27.9)μm。未观察到卵孢子和厚垣孢子。将同一分离株与来自冬青栎(PH050,交配型A1)和乳香黄连木(PH017,交配型A2)的两个隐地疫霉分离株在胡萝卜琼脂上于23±1°C黑暗条件下进行配对试验。只有DB13GIU02与PH017的配对成功,产生了直径为28.3至34.6(平均31.7)μm的藏卵器、直径为28.0至32.2(平均29.2)μm的卵孢子以及大小为10.5至15.1×11.6至15.1(平均13.5×13.3)μm的围生雄器。使用Nucleospin Plant试剂盒(Macherey Nagel)提取三个分离株的DNA。用引物Cryp 1和Cryp 2(4)对所有隐地疫霉分离株的DNA进行PCR扩增,产生了一个特异性扩增子。使用引物ITS1/ITS4扩增分离株DB13GIU02的rDNA内部转录间隔区(ITS)并进行测序。对845 bp片段(GenBank登录号KM458193)的BLAST分析显示与隐地疫霉GU111631的序列有99%的同源性。通过使用在2:1小麦/麻籽混合物上生长的一株隐地疫霉对60日龄健康鼠尾草植株进行致病性试验。将受侵染的种子(10 g/升基质)混入基于水藓泥炭/腐叶土/松树皮/粘土(50:20:20:10 v/v)的蒸汽消毒基质中。对照植株用混入蒸汽消毒土壤中的未接种小麦/麻籽处理。试验重复一次。每个处理使用15株植物。所有植物置于20±1°C的生长室中。接种植株在接种后7天出现黄化,接种后15天出现根腐和冠腐。持续从接种植株中重新分离出隐地疫霉。在选择性培养基上未从无症状的对照植株中分离到菌落。在美国已报道隐地疫霉侵染鼠尾草(2),而在意大利,在白花鼠尾草上观察到了相同病原菌。这是意大利首次报道隐地疫霉侵染鼠尾草。由于这种植物作为烹饪用香料和园林植物的用途不断扩大,该病的经济重要性可能会增加。参考文献:(1)D. C. Erwin和O. K. Ribeiro。《全球疫霉病害》。美国植物病理学会出版社,明尼苏达州圣保罗,1996年。(2)S. T. Koike等人。《植物病害》81:959,1997年。(3)H. Masago等人。《植物病理学》67:25,1977年。(4)D. Minerdi等人。《欧洲植物病理学杂志》122:227,2008年。

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