Symptomatic sweet potato cv Arapey INIA samples were collected from a commercial production field in Colonia Molina, Guaymallén department, Mendoza province, Argentina. They showed dark rounded lesions, sometimes coalescing with white granular mycelium. Fungus was obtained from symptomatic sweet potatoes, which represented the generalized infection that affected the crop. They were seeded in PDA with streptomycin sulfate and incubated for seven days at 21°C, alternating white/black (UV400nm) light. Observations with an optical microscope revealed the presence of hyaline, not septated, cylindrical endoconidia with rounded ends. They were 8-16 μm length and 4–6 μm width. Phialides were 43-46 μm length, rounded bases (7-9 μm width) and tapering to the neck´s tip (4-6 μm width). Brown chlamydospores (aleuriospores), 9-13 μm length and 8-12 μm width, in chains of 2-8 spores were observed. For molecular identification, total genomic DNA was extracted. ITS fragment of 565 pb was amplified using ITS5/ITS4 primers and sequenced. The sequence indicated 99% identity with Berkeleyomyces basicola (synonymous: Thielaviopsis basicola). This was deposited in GenBank as (KX580957) (CBS: C430.74, Gen Bank accession number AF275482.1). This is the first report of B. basicola in sweet potato in Argentina, a potential threat to storage root yields. Highlights: Sweet potato black root rot, new disease in Argentina. First report of Berkeleyomyces basicola causing black root rot on sweet potato in Mendoza, Argentina.
13The soil-borne fungal plant pathogen Verticillium dahliae can infect more than 300 plant 14 species including important economic crops, causing great economic loses. V. dahliae 15 can persist and survive more than 14 years in the soil by resistance structures, known as 16 microsclerotia, which constitute the primary inoculum in the field. In vitro mass 17 production of microsclerotia is essential for performing many pathological assays. 18Nevertheless to harvest the microsclerotia is not an easy task and several protocols have 19 been described although none of them is completely satisfying for different reasons. here 20 we present a new protocol that is reproducible, robust, simple and fast allows to overcome 21 the difficulties for obtaining massive amounts of microsclerotia. In summary, we 22 developed a new culture medium that we called Pluronic Potato Medium (PPM) because 23 it is essentially potato dextrose media with the hydrogel, Pluronic F127 as a solidifying 24 agent. The microsclerotia collected in form PPM were infectious in tomato plants were 25 2 they were able to reproduce the disease and we recovered and quantitated V. dahliae in 26 infected plants. 27Keywords Verticillum dahliae, microsclerotia, inoculum, mass production, pluronic 28 29 This difficulty was absolutely overcomed by using a thermoreversible hydrogel as 175 Pluronic F-127 as a solidifying agent in PPM medium. 176 Poloxamer or Pluronics hydrogels are triblock copolymers (nonionic, polyoxyethylene-177 polyoxypropylene-polyoxyethylene (PEO-PPO-PEO)) that have multiple pharmaceutical 178 applications (Bohorquez et al., 1999). A widely explored member of this family is 179 Pluronic F127, which has 1200 kDa PPO units and 70% PEO content (Wang et al., 2019).180The most striking feature of this hydrogel is that, due to its composition, it shows 181 transformations as a function of temperature changes. This characteristic of 182 thermoreversible gel is useful in concentrations higher than or equal to 20%. Cold 183 temperatures cause the gel to go from solid to liquid state, while remaining solid at room 184 temperature (Akash & Rehman, 2015). These characteristics of appropriate hydrophilic / 185 lipophilic balance, high molecular weight, thermos-reversibility and lower mechanical 186 strength allow for a high extraction capacity in aqueous phase (Lv et al. 2007) by TIPS 187 (Qiu et al., 2008). Hydrogels such as Pluronic F127 have been used mainly as detergents, 188 stabilizers, foaming agents and emulsifiers in the field of biomedicine, in tissue 189 engineering and pharmacology (Bercea et al., 2011; Gwon et al., 2018; Wu et al., 2018). 190 Recently, they have been used in studies of interactions between plant parasitic nematodes 191 and plants (Li et al., 2019). 192 The main advantages of a massive production of V. dalihae MS using the PPM media and 193 the methodology here described are the short time elapsed from inoculating the mycelium 194 to the moment of collecting the MS, the minimal physical space needed for producing 195 MS an...
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