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南卡罗来纳州由齐整小核菌引起的葫芦(Lagenaria siceraria)白绢病的首次报道。

First Report of Southern Blight on Bottle Gourd (Lagenaria siceraria) Caused by Sclerotium rolfsii in South Carolina.

作者信息

Ling K S, Kousik C S, Keinath A P

机构信息

USDA, Agricultural Research Service, U.S. Vegetable Laboratory, Charleston, SC.

Clemson University, Coastal Research and Education Center, Charleston, SC.

出版信息

Plant Dis. 2008 Apr;92(4):656. doi: 10.1094/PDIS-92-4-0656C.

Abstract

Bottle gourd (Lagenaria siceraria (Mol.) Standl.) is an important rootstock in watermelon production in several countries such as Japan, China, and Israel where 60 to 70% of watermelons are grafted (2). We are evaluating bottle gourds for their ability to improve disease resistance when used as rootstock for watermelon (3). In the summer of 2007, symptoms of wilting and crown necrosis appeared on bottle gourd seedlings 1 month after transplanting in a field in Charleston, SC. Infection was observed on commercial cv. Emphasis and four advanced breeding lines. In October of 2007, 35 of 85 plants examined (41%) had stem rot at the crown area just above the soil line where coarse, white mycelia and abundant sclerotia were observed. The fungus tentatively identified as Sclerotium rolfsii produced sclerotia that were white or light to dark brown and measured 0.6 to 2.5 mm in diameter (mean = 1.1 mm). Diseased tissues with sclerotia from four plants were disinfested for 1 min in 0.5% sodium hypochlorite and plated on acidified potato dextrose agar (APDA). Fungal colonies that produced white mycelia and tan-to-brown sclerotia were isolated from four wilted plants. A single PCR product of approximately 680 bp was amplified from DNA extracted from two isolates using the primers ITS1 and ITS4 (4). One PCR product was cloned into the TOPO TA cloning vector (Invitrogen, Carlsbad, CA) and sequenced (GenBank Accession No. EU338381). BLASTN analysis of the sequence in the NCBI databases revealed 99% similarity to the internal transcribed spacer (ITS) sequences of S. rolfsii and Athelia rolfsii (perfect stage of S. rolfsii), confirming that the pathogen was indeed S. rolfsii. Two S. rolfsii isolates were used to test pathogenicity. Each isolate was used to inoculate five young seedlings and five adult (10-week-old) bottle gourd plants. For inoculation, 10 sclerotia obtained from the APDA plates were placed on the surface of the potting soil 0.5 to 1 cm from the collar region of each bottle gourd plant growing in 10-cm pots. Inoculations were done carefully to ensure that the plants were not injured. After inoculation, the plants were maintained at high humidity and 25°C for 3 days and then transferred to laboratory benches. Four young seedlings and three adult noninoculated plants kept under the same conditions served as controls. The pathogenicity test was repeated once with similar results. All inoculated plants developed symptoms of southern blight. The inoculated plants developed symptoms of wilting 4 to 5 days after inoculation and completely wilted within 7 to 10 days. Symptoms of wilting were soon followed by the appearance of white-to-light brown sclerotia on the collar region. No symptoms were observed on the noninoculated plants. S. rolfsii was reisolated from the inoculated plants on APDA. Although southern blight caused by S. rolfsii has been reported on many crop plants in the southern United States, to our knowledge, this disease has not been reported previously on bottle gourd in North America. However, the disease has been reported on bottle gourd in India (1). Identifying sources of resistance to southern blight in bottle gourds may be necessary to make them suitable as rootstocks in areas where S. rolfsii is present. References: (1) K. S. Amin. Indian Phytopathol. 34:253, 1981. (2) R. Cohen et al. Plant Dis. 91:916, 2007. (3) K. S. Ling and A. Levi. HortScience 42:1124, 2007. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Amplifications. Academic Press, San Diego, 1990.

摘要

瓠瓜(Lagenaria siceraria (Mol.) Standl.)是日本、中国和以色列等几个国家西瓜生产中的一种重要砧木,在这些国家,60%至70%的西瓜是嫁接的(2)。我们正在评估瓠瓜作为西瓜砧木时提高抗病性的能力(3)。2007年夏天,在南卡罗来纳州查尔斯顿的一块田地中,瓠瓜幼苗在移栽1个月后出现萎蔫和冠部坏死症状。在商业品种Emphasis和4个先进育种系上观察到了感染情况。2007年10月,在检查的85株植株中,有35株(41%)在土壤线以上的冠部区域发生茎腐病,在那里观察到粗糙的白色菌丝体和大量菌核。初步鉴定为齐整小核菌的真菌产生的菌核为白色或浅至深褐色,直径为0.6至2.5毫米(平均 = 1.1毫米)。从4株带有菌核的患病组织在0.5%次氯酸钠中消毒1分钟,然后接种在酸化马铃薯葡萄糖琼脂(APDA)上。从4株萎蔫植株上分离出产生白色菌丝体和棕褐色菌核的真菌菌落。使用引物ITS1和ITS4从两个分离株提取的DNA中扩增出一个约680 bp的单一PCR产物(4)。一个PCR产物被克隆到TOPO TA克隆载体(Invitrogen,卡尔斯巴德,加利福尼亚州)中并进行测序(GenBank登录号EU338381)。在NCBI数据库中对该序列进行BLASTN分析,结果显示与齐整小核菌和罗氏阿太菌(齐整小核菌的有性阶段)的内部转录间隔区(ITS)序列有99%的相似性,证实该病原菌确实是齐整小核菌。使用两个齐整小核菌分离株进行致病性测试。每个分离株用于接种5株幼苗和5株成年(10周龄)瓠瓜植株。为了接种,从APDA平板上获得的10个菌核被放置在种植在10厘米花盆中的每株瓠瓜植株颈部区域0.5至1厘米处的盆栽土壤表面。接种时小心操作以确保植株不受损伤。接种后,植株在高湿度和25°C条件下保持3天,然后转移到实验室工作台上。4株未接种的幼苗和3株成年植株在相同条件下作为对照。致病性测试重复一次,结果相似。所有接种的植株都出现了白绢病症状。接种的植株在接种后4至5天出现萎蔫症状,并在7至10天内完全萎蔫。萎蔫症状出现后不久,在颈部区域出现白色至浅褐色菌核。未接种的植株未观察到症状。从接种的植株上在APDA上重新分离出齐整小核菌。尽管在美国南部许多作物上都报道过由齐整小核菌引起的白绢病,但据我们所知,这种病害以前在北美的瓠瓜上尚未报道。然而,在印度的瓠瓜上已报道过这种病害(1)。确定瓠瓜对白绢病的抗性来源对于使其适合在存在齐整小核菌的地区作为砧木可能是必要的。参考文献:(1)K. S. Amin。《印度植物病理学》34:253,1981。(2)R. Cohen等人。《植物病害》91:916,2007。(3)K. S. Ling和A. Levi。《园艺科学》42:1124,2007。(4)T. J. White等人。《PCR协议:方法与扩增指南》。学术出版社,圣地亚哥,1990。

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