Garibaldi A, Gilardi G, Ortu G, Gullino M L, Testa M
Centre of Competence for Innovation in the Agro-Environmental Sector (AGROINNOVA), Via Leonardo da Vinci 44, 10095 Grugliasco, Italy.
AGROINNOVA and DISAFA, Via Leonardo da Vinci, 44, 10095 Grugliasco, Italy.
Plant Dis. 2013 Oct;97(10):1386. doi: 10.1094/PDIS-12-12-1165-PDN.
Common bean (Phaseolus vulgaris L.) is grown worldwide for consumption of dry or green beans. During late spring of 2012, yellowing and wilting symptoms were observed in a commercial bean field cv. Lingua di fuoco in Cagliari Province (Sardinia, southern Italy) on 30% of plants 4 to 5 months after sowing. The first symptoms developed in May, when temperatures reached 18 to 30°C. Affected plants showed crown rot, necrosis of the cortex, and foliar chlorosis. As disease progressed, plants collapsed. In the presence of abundant moisture, white mycelium developed on the senescent tissue along with light to dark brown sclerotia (3.0 to 4.8 mm in diameter). Symptomatic tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 25 mg streptomycin sulfate/liter. The fungus that was isolated consistently from symptomatic plants onto PDA at 23°C grew rapidly in culture with silky-white, sterile mycelium, formed light to dark brown sclerotia (each 1.8 to 3.2 mm in diameter) after 7 days, and readily produced aerial hyphae. These morphological features are typical of Sclerotium rolfsii (2). The internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) was amplified for one isolate using ITS1/ITS4 primers (4), and sequenced (GenBank Accession No. KF002510). BLASTn analysis (1) of the 656-bp segment showed 87% homology with the ITS sequence of an S. rolfsii isolate (JF819727). Pathogenicity of one isolate was confirmed by inoculating healthy P. vulgaris plants cv. Lingua di fuoco grown in 2-liter pots in a steamed potting mix containing 50% Tecno2 (70% white peat and 30% clay) and 50% Tiesse 3 (60% white peat, 20% clay, and 20% perlite) (Turco Silvestro terricci, Bastia d'Albenga, SV, Italy). Inoculum consisting of mycelium and sclerotia of the pathogen produced from 10-day-old cultures on PDA was mixed in the soil at 0.5 g/liter substrate. Four 7-day-old plants per pot, with three replicate pots, were used for inoculation. The same number of control plants grown in the same substrate were inoculated with non-colonized PDA as a negative control treatment. The pathogenicity test was repeated. Plants were kept in a growth chamber at 30°C and 85% RH. Inoculated plants developed symptoms of leaf yellowing within 10 days, followed by crown rot, appearance of white mycelium and sclerotia, and eventual wilting. Control plants remained asymptomatic. Isolations from inoculated plants demonstrated the absence of latent infections by the fungus S. rolfsii, but the fungus was not reisolated from non-inoculated control plants. To our knowledge, this is the first report of S. rolfsii infecting P. vulgaris in Italy. Southern blight has been reported on common bean in sub-tropical and tropical areas of the world (3), where it can cause severe crop losses. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) J. E. M. Mordue. CMI Descriptions of Pathogenic Fungi and Bacteria No. 410, 1974. (3) H. F. Schwartz et al. Page 20 in: Compendium of Bean Diseases. American Phytopathological Society Press, St. Paul, MN, 2005. (4) T. J. White et al. PCR Protocols. Page 315 in: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.
菜豆(Phaseolus vulgaris L.)在全球范围内种植,用于食用干豆或嫩豆。2012年春末,在撒丁岛(意大利南部)卡利亚里省一片种植商业品种“Lingua di fuoco”的菜豆田中,播种4至5个月后,30%的植株出现了黄化和萎蔫症状。最初症状出现在5月,当时气温达到18至30°C。受影响的植株表现出根颈腐烂、皮层坏死和叶片黄化。随着病情发展,植株枯萎。在湿度充足的情况下,衰老组织上长出白色菌丝体,并伴有浅褐色至深褐色菌核(直径3.0至4.8毫米)。将有症状的组织在1%次氯酸钠中消毒1分钟,然后接种到添加了25毫克/升硫酸链霉素的马铃薯葡萄糖琼脂(PDA)平板上。在23°C条件下,从有症状植株上 consistently 分离到PDA上的真菌在培养中生长迅速,形成丝滑白色的无菌菌丝体,7天后形成浅褐色至深褐色菌核(每个直径1.8至3.2毫米),并易于产生气生菌丝。这些形态特征是齐整小核菌(Sclerotium rolfsii)的典型特征(2)。使用ITS1/ITS4引物(4)对一个分离株的核糖体DNA(rDNA)的内部转录间隔区(ITS)进行扩增,并进行测序(GenBank登录号KF002510)。对656碱基片段的BLASTn分析(1)显示,与齐整小核菌一个分离株(JF819727)的ITS序列有87%的同源性。通过将在PDA上培养10天的病原菌的菌丝体和菌核组成的接种物以0.5克/升基质的量混入土壤中,对接种健康的“Lingua di fuoco”菜豆植株(种植在装有50%Tecno2(70%白色泥炭和30%粘土)和50%Tiesse 3(60%白色泥炭、20%粘土和20%珍珠岩)的2升花盆中,Turco Silvestro terricci,巴斯蒂亚-达尔本加,SV,意大利)来确认一个分离株的致病性。每个花盆种植4株7日龄植株,设置三个重复花盆用于接种。在相同基质中生长的相同数量的对照植株接种未被定殖的PDA作为阴性对照处理。重复进行致病性试验。将植株置于温度30°C、相对湿度85%的生长室中。接种植株在10天内出现叶片黄化症状,随后出现根颈腐烂、白色菌丝体和菌核出现,最终萎蔫。对照植株无症状。对接种植株的分离表明不存在齐整小核菌的潜伏感染,但未从未接种的对照植株中重新分离到该真菌。据我们所知,这是齐整小核菌在意大利感染菜豆的首次报道。南方疫病在世界亚热带和热带地区的菜豆上已有报道(3),在这些地区它可导致严重的作物损失。参考文献:(1)S. F. Altschul等人,《核酸研究》25:3389,1997。(2)J. E. M. Mordue,《英联邦真菌病原和细菌描述》第410号,1974。(3)H. F. Schwartz等人,《菜豆病害简编》第20页,美国植物病理学会出版社,圣保罗,明尼苏达州,2005。(4)T. J. White等人,《PCR协议》,《方法与应用指南》第315页,学术出版社,圣地亚哥,加利福尼亚州,1990。