Garibaldi A, Gilardi G, Gullino M L
Centre of Competence for the Innovation in the Agro-Environmental Sector (AGROINNOVA), Via Leonardo da Vinci 44, 10095 Grugliasco, Italy.
Plant Dis. 2010 Dec;94(12):1504. doi: 10.1094/PDIS-03-10-0206.
Witloof chicory (Cichorium intybus L.) is an important crop in Italy where most of the crop is still produced in soil. In September 2009, chicory plants (cv. Pan di Zucchero) grown on a commercial farm in Tarquinia (central Italy) showed symptoms of a previously unknown disease. Symptoms, observed 20 days after transplanting, consisted of stunting, yellowing of leaves, and a crown and root rot. Affected plants turned brown, wilted, and eventually died. At the soil level, dark brown-to-black water-soaked lesions coalesced and often girdled the stem. All of the crown and root system was affected. At this location, the disease was severe and widespread, with 60% of observed plants being affected. A Phytophthora-like organism was consistently isolated on a medium selective for oomycetes (4) after disinfestation of lower stem and root pieces of C. intybus for 1 min in a solution containing 1% NaOCl. Tissue fragments of 1 mm were excised from the margins of the root and crown lesions. The pathogen genus was identified as Phytophthora based on morphological and physiological features. Sporangia were produced for identification by growing a pure culture for 15 days on modified V8 juice agar medium (Campbell V8 juice [200 ml], agar [15 g], CaCo [0.5 g], and sterile water [800 ml]) under alternating light and dark (12/12 h). Sporangia were pyriform to ovoid, papillate, and measured 33.3 to 59.2 × 18.9 to 30.2 μm (average 39.9 × 25.8 μm). Chlamydospores developed in 28-day-old cultures and measured 21.3 to 30.2 × 19.5 to 29.7 μm (average 24.4 × 23.6 μm). Oogonia were globose and measured 26 to 41 μm (average 32.5 μm). Eighty percent of antheridia were paragynous. Amphyginous antheridia (15 to 20%) were also observed. Oospores were scarcely produced and measured 24 to 32 μm in diameter. The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 851-bp segment showed 100% homology with the sequence of Phytophthora tentaculata. The nucleotide sequence has been assigned GenBank Accession No. GU949536. Pathogenicity of this isolate was confirmed by inoculating C. intybus cv. Pan di Zucchero plants 20 days after transplant. The same isolate was grown for 15 days on a mixture of 70:30 wheat/hemp kernels and then 5 g/liter of the inoculum was mixed into a substrate containing a mixture of blond and black peat (15:85 vol/vol), pH 5.5. Five plants per 2-liter pot were transplanted and four replicates were carried out. Twenty noninoculated plants represented the control treatment. The trial was repeated. Plants were kept in two growth chambers at two temperatures (20 and 25°C). Symptoms similar to those observed in the field developed 7 days after inoculation. Twenty days later, 100 and 40% of the plants were dead at 25 and 20°C, respectively. Control plants remained symptomless. P. tentaculata was consistently reisolated from symptomatic plants. To our knowledge, this is the first report of P. tentaculata on C. intybus in the world ( http://nt.ars-grin.gov/fungaldatabases/index.cfm ). P. tentaculata was recently reported on lavender in Spain (2) and oregano in Italy (3). The economic importance of this disease is relatively low on most commercial farms. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) L. Alvarez et al. Plant Dis. 90:523, 2006. (3) P. Martini et al. Plant Dis. 93:843, 2009. (4) H. Masago et al. Phytopathology 67:425, 1977.
菊苣(菊苣属)是意大利的一种重要作物,该国大部分菊苣仍在土壤中种植。2009年9月,在塔尔奎尼亚(意大利中部)一个商业农场种植的菊苣植株(品种为“Pan di Zucchero”)出现了一种此前未知疾病的症状。移栽20天后观察到的症状包括植株矮小、叶片发黄以及根茎腐烂。受影响的植株变为褐色,枯萎,最终死亡。在土壤层面,深褐色至黑色的水渍状病斑融合,常常环绕茎部。整个根茎系统均受影响。在此处,该病严重且广泛传播,观察到的植株中有60%受到影响。在将菊苣的茎基部和根部组织块在含1%次氯酸钠的溶液中消毒1分钟后,在一种对卵菌有选择性的培养基上持续分离出一种类似疫霉的生物体。从根茎病斑边缘切下1毫米的组织碎片。基于形态和生理特征,该病原菌被鉴定为疫霉属。通过在改良V8汁琼脂培养基(坎贝尔V8汁[200毫升]、琼脂[15克]、碳酸钙[0.5克]和无菌水[800毫升])上在交替光照和黑暗(12/12小时)条件下培养纯培养物15天来产生孢子囊用于鉴定。孢子囊呈梨形至卵形,具乳突,大小为33.3至59.2×18.9至30.2微米(平均39.9×25.8微米)。在培养28天的培养物中产生厚垣孢子,大小为21.3至30.2×19.5至29.7微米(平均24.4×23.6微米)。藏卵器呈球形,大小为26至41微米(平均32.5微米)。80%的雄器为侧生。也观察到周生雄器(15%至20%)。卵孢子很少产生,直径为24至32微米。使用引物ITS4/ITS6扩增单个分离物的rDNA的内部转录间隔区(ITS)并进行测序。对851碱基对片段的BLAST分析显示与触手疫霉的序列有100%的同源性。该核苷酸序列已被赋予GenBank登录号GU949536。通过在移栽20天后接种菊苣品种“Pan di Zucchero”植株来确认该分离物的致病性。将同一分离物在70:30的小麦/麻籽混合物上培养15天,然后将5克/升的接种物混入含有金发藓泥炭和黑泥炭混合物(15:85体积/体积)、pH 5.5的基质中。每2升花盆移栽5株植物,进行4次重复。20株未接种的植物作为对照处理。该试验重复进行。植物置于两个生长室中,温度分别为20和25°C。接种7天后出现与在田间观察到的类似症状。20天后,在25°C和20°C下分别有100%和40%的植物死亡。对照植物无症状。从有症状的植物中持续重新分离到触手疫霉。据我们所知,这是世界上关于触手疫霉侵染菊苣的首次报道(http://nt.ars-grin.gov/fungaldatabases/index.cfm)。触手疫霉最近在西班牙的薰衣草(2)和意大利的牛至(3)上有报道。这种病害在大多数商业农场的经济重要性相对较低。参考文献:(1)S.F.Altschul等人,《核酸研究》25:3389,1997。(2)L.Alvarez等人,《植物病害》90:523,2006。(3)P.Martini等人,《植物病害》93:843,2009。(4)H.Masago等人,《植物病理学》67:425,1977。