First report of Cladosporium cladosporioides as causal agent of leaf blight on Borassus flabellifer in India
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First report of Cladosporium cladosporioides as causal agent of leaf blight on Borassus flabellifer in India Arghya Banerjee 1 & Pradip Sarkar 1 & Birendranath Panja 1 & Partha Sarathi Nath 1 Received: 3 August 2019 / Accepted: 30 January 2020 # Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2020
Keywords Cladosporium cladosporioides species complex . EF1-α . ITS barcode . Morphology . Pathogenicity . Phylogenetic analysis
Borassus flabellifer L. (Arecaceae), commonly known as toddy palm, is a popular robust fruit tree native to Indian subcontinent and Southeast Asia. Its leaves, fruits, sprouts and trunks have immense economical values in Indian culture (Gummadi et al. 2016). During end of December 2018, a severe foliar blight was observed on road-site plantations of B. flabellifer located at Itachuna, Hooghly district of West Bengal. Disease incidence ranged from 20% to 50% in some areas. Initially small to medium sized streaks were found parallel to vein or marginal leaf regions, and finally coalesced and whole leaves looked like a blighted appearance. Dark brown to black dusty pustules were found on the surface of blighted leaf areas. Symptomatic leaves were superficially disinfected using 0.1% HgCl2 for one minute, placed on potato dextrose agar (PDA), and incubated at 25 °C for five days. Colonies on PDA were olive-green to brown and showed a velvety texture. Conidiophores were pale and smooth or sometimes verruculose (141.3 μm [60 to 250 μm] × 3.8 μm [3.2 to 4 μm]) (n = 30). Ramoconidia were 0- to 1-septate and were smooth or sometimes minutely verruculose (16 μm [12 to 30 μm] × 2.8 μm [2 to 4 μm]) (n = 30). Conidia formed in long and branched chains that were readily disarticulate, single celled, and elliptical to limoniform (4.6 μm [2 to 9 μm] × 2.2 μm [2 to 3 μm]) (n = 50). Conidia were pale- to olive-brown and Arghya Banerjee and Partha Sarathi Nath contributed equally to this work. * Arghya Banerjee [email protected] 1
Department of Plant Pathology, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal 741252, India
smooth to verruculose. Based on these morphological features the fungus was identified as Cladosporium cladosporioides (Fresen.) G.A. de Vries. For molecular identification, the internal transcribed spacer (ITS) region and a portion of translation elongation factor 1-α (EF1-α) gene were amplified by PCR and sequenced using the primer sets ITS1/ITS2 (GenBank Accession No. MK492259) and EF1-728F/EF1-986R (MK495431), respectively. BLAST analysis showed in both cases 100% identity with existing sequences of C. cladosporioides (MN503474; MG775038). The phylogenetic analysis resulted in a well-supported clade grouped with the type species of C. cladosporioides. Based on the morphological characteristics and the molecular data, the causal agent of observed leaf blight symptoms was identified as C. cladosporioides (Bensch et al. 2012). Pathogenicity of the fungus was tested on superficially disinfected six leaves of one-year-old B. flabellifer plants by sp
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