First Report of Leaf Spot of Spinach (Spinacia oleracea) Caused by Myrothecium verrucaria in Italy

Abstract

HomePlant DiseaseVol. 100, No. 8First Report of Leaf Spot of Spinach (Spinacia oleracea) Caused by Myrothecium verrucaria in Italy PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Leaf Spot of Spinach (Spinacia oleracea) Caused by Myrothecium verrucaria in ItalyA. Garibaldi, G. Gilardi, S. Franco-Ortega, and M. L. GullinoA. GaribaldiSearch for more papers by this author, G. GilardiSearch for more papers by this author, S. Franco-OrtegaSearch for more papers by this author, and M. L. GullinoSearch for more papers by this authorAffiliationsAuthors and Affiliations A. Garibaldi G. Gilardi S. Franco-Ortega M. L. Gullino , Centre of Competence for the Agro-Environmental Sector (AGROINNOVA) and DISAFA, Largo P. Braccini 2, 10095 Grugliasco, Italy. Published Online:24 May 2016https://doi.org/10.1094/PDIS-01-16-0012-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat During the spring of 2015, plants of spinach (Spinacia oleracea L.) ‘Donkey’ were grown using conventional control strategies in plastic houses in the Lumbardy Region near Brescia (Northern Italy). The plants showed symptoms of a previously unknown foliar disease on 15- to 20-day-old plants that consisted of small, circular, sunken, gray-brown spots (1 to 2 mm in diameter) with a well-defined border. As the lesions expanded (up to 30 mm in diameter) they developed concentric rings, coalesced, and then the affected tissues became water-soaked. Creamy to black sporodochia appeared on the affected leaves. Roots were asymptomatic. Five hectares of plastic houses irrigated with well water by overhead sprinklers were affected, with 5 to 10% of plants exhibiting symptoms. Affected plants showed 30 to 60% incidence of diseased leaves. Isolations were carried out on symptomatic leaf-tissue sections (each 1 mm2) dipped in 1% sodium hypochlorite for 1 min, rinsed in sterilized water, dried on sterilized filter paper, and then placed on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. A white slow-growing fungus was obtained that developed black sporodochia 7 to 10 days after plating, similar to those present on the affected leaves. Conidia were cylindrical, nonseptate, without appendages, measuring 5.8 to 9.2 and 1.9 to 2.8 (avg. of 30 conidia, 7.6 ± 1.14 × 2.4 ± 0.22) µm. The morphological characteristics of the isolated pathogen corresponded to those of Myrothecium spp. (Domsch et al. 1980). The Internal Transcribed Spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis (Altschul et al. 1997) of the 516-bp segment showed a 100% similarity with M. verrucaria KR708633.1. A GenBank Accession No. KT354922 was assigned. To confirm pathogenicity, 15-day-old leaf spinach plants cv. Donkey were transplanted in 2-liter pots, filled with a steamed peat, perlite, and sand substrate in a ratio of 60:20:20 vol/vol and maintained in a growth chamber between 20 and 24°C. Five pots per treatment were used with five plants per pot. The artificial inoculation was carried out by spraying leaves with a spore suspension (1 × 105 conidia/ml) prepared from 15-day-old PDA cultures of one representative isolate of the pathogen grown under 12 h of photoperiod at 20°C. Control plants were inoculated with distilled water. Plants were kept covered with plastic bags for 5 days. Spots similar to those of the original plants developed 7 days after the inoculation, with a disease incidence ranging from 60 to 70% of the plants affected. All the noninoculated plants remained asymptomatic. A fungus morphologically identified as M. verrucaria was consistently isolated from all the symptomatic plants. The pathogenicity test was conducted twice, showing the same results. This is the first report of M. verrucaria on S. oleracea in Italy as well as worldwide. The cultivation of spinach for the fresh market is increasing in recent years in northern Italy. Currently, this disease has spread to several farms in the Lumbardy Region, causing limited crop losses.