Garibaldi A, Bertetti D, Pensa P, Gullino M L
Center of Competence AGROINNOVA, University of Torino, Via Leonardo da Vinci, 44, 10095 Grugliasco, Italy.
Plant Dis. 2010 Jan;94(1):129. doi: 10.1094/PDIS-94-1-0129B.
Lampranthus spp. N.B. Brown (figmarigold) belongs to the Aizoaceae family and is grown as a ground cover in gardens or as a potted plant. In January 2009, severe outbreaks of a previously unknown wilt were observed at a commercial farm in Liguria (northern Italy) where 7-month-old potted plants were grown outdoors in a mix of peat/clay/pumice at pH 6.5. In cultivars with pink flowers, 12% of plants were affected, while only a few cultivars with red flowers were diseased. Initial symptoms were yellowing of external leaves and brown or black streaks in the vascular tissue of roots, crown, and leaves. Subsequently, infected tissues wilted and stopped growing, stems and leaves appeared desiccated, and infected plants died. Stems of 10 pink-flowered plants were severed with a knife, cut ends sealed with wax, and surfaces disinfected with 1% sodium hypochlorite. Cross-sections (1 mm long) through symptomatic vascular tissue were plated onto potato dextrose agar. After 10 days at 22°C, 90% of the stems tested positive for Verticillium. Irregular, dark microsclerotia, 22 to 128 × 13 to 66 (average 51 × 29) μm, developed in hyaline hyphae after 10 days of growth at 22 ± 1°C (12-h photoperiod). Hyaline, elliptical, single-celled conidia, 2.9 to 4.8 × 1.3 to 2.4 (average 4.1 × 1.7) μm, developed on verticillate conidiophores. On the basis of these morphological characteristics, the fungus was identified as V. dahliae (3). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 (2) and sequenced. BLASTn analysis (1) of the 476-bp segment showed a 100% homology with the sequence of V. dahliae. The nucleotide sequence has been assigned GenBank Accession No. GQ 149479. Pathogenicity tests were performed twice using five 40-day-old plants of a pink-flower cultivar of a Lampranthus sp. grown in 1-liter pots containing a 50:20:20:10 mix of peat moss/pumice/pine bark cortex/clay. The substrate was infested with a conidial suspension (1.0 × 10/ml) of one isolate of V. dahliae recovered from infected plants. Inoculum (50 ml) were added to each pot, drenching the top of the soil. Noninoculated plants served as controls. Plants (five per treatment) were maintained in a glasshouse at daily average temperatures between 20 and 26°C and at 50 to 70% relative humidity. The first wilt symptoms and a vascular discoloration in the roots, crown, and veins developed 30 days after inoculation. V. dahliae was consistently reisolated. Noninoculated plants remained healthy. In a second test, the susceptibility of purple-, white-, yellow-, red-, and orange-flowered cultivars was tested. Ten rooted cuttings of each cultivar were inoculated as described above. The severity of Verticillium wilt was evaluated and each cultivar was classified as resistant, partially resistant, average susceptible, susceptible, or highly susceptible. All tested cultivars were susceptible or highly susceptible to Verticillium. Only the purple cultivar showed an average susceptibility. To our knowledge, this is the first report of Verticillium wilt on Lampranthus spp. in Italy as well as worldwide. Today, the economic importance of Verticillium wilt on figmarigold in Italy is still limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. A. Innis et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990. (3) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002.
松叶菊属植物(松叶菊)N.B. 布朗属于番杏科,作为地被植物种植于花园中或作为盆栽植物。2009年1月,在利古里亚(意大利北部)的一个商业农场观察到一种此前未知的枯萎病严重爆发,那里7个月大的盆栽植物种植在pH值为6.5的泥炭/黏土/浮石混合基质中并置于户外。在开粉色花的品种中,12%的植株受到影响,而只有少数开红色花的品种染病。最初症状是外部叶片发黄以及根、茎基部和叶片维管组织出现褐色或黑色条纹。随后,受感染组织枯萎并停止生长,茎和叶出现干枯,受感染植株死亡。用刀切断10株开粉色花植株的茎,切口用蜡密封,表面用1%次氯酸钠消毒。将有症状的维管组织切成1毫米长的横切片接种到马铃薯葡萄糖琼脂培养基上。在22°C下培养10天后,90%的茎对大丽轮枝菌检测呈阳性。在22±1°C(12小时光周期)生长10天后,在透明菌丝中形成不规则的黑色微菌核,大小为22至128×13至66(平均51×29)微米。透明、椭圆形、单细胞分生孢子,大小为2.9至4.8×1.3至2.4(平均4.1×1.7)微米,着生于轮状分生孢子梗上。基于这些形态特征,该真菌被鉴定为大丽轮枝菌(3)。使用引物ITS4/ITS6(2)扩增rDNA的内部转录间隔区(ITS)并进行测序。对476碱基对片段的BLASTn分析(1)显示与大丽轮枝菌的序列有100%的同源性。该核苷酸序列已被赋予GenBank登录号GQ 149479。使用5株40日龄的开粉色花的松叶菊属品种植株进行了两次致病性试验,植株种植在1升花盆中,基质为泥炭藓/浮石/松树皮皮层/黏土按50:20:20:10混合而成。基质用从受感染植株分离得到的一株大丽轮枝菌的分生孢子悬浮液(1.0×10/毫升)侵染。向每个花盆中加入50毫升接种物,浇灌土壤表层。未接种的植株作为对照。植株(每个处理5株)置于温室中,日平均温度在20至26°C之间,相对湿度在50至70%。接种30天后出现首次枯萎症状以及根、茎基部和叶脉的维管变色。持续重新分离得到大丽轮枝菌。未接种的植株保持健康。在第二次试验中,测试了开紫色、白色、黄色、红色和橙色花的品种的易感性。每个品种取10个带根插条按上述方法接种。评估了黄萎病的严重程度,每个品种被分类为抗病、部分抗病、中度感病、感病或高度感病。所有测试品种对大丽轮枝菌均感病或高度感病。只有紫色品种表现出中度感病。据我们所知,这是意大利以及全球关于松叶菊属植物黄萎病的首次报道。如今,黄萎病在意大利对松叶菊的经济重要性仍然有限。参考文献:(1)S.F. 阿尔茨舒尔等人,《核酸研究》25:3389,1997年。(2)M.A. 英尼斯等人,见《PCR协议:方法与应用指南》第315页。学术出版社,加利福尼亚州圣地亚哥,1990年。(3)G.F. 佩格和B.L. 布雷迪,《黄萎病》。CABI出版社,英国沃灵福德,2002年。