Indian Phytopath. 70 (3) : 326-330 (2017) DOI 10.24838/ip.2017.v70.i3.72493 Histopathological and molecular characterization of Ustilaginoidea virens in rice POOJA KUMARI* and R.K. SHARMA Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi 110 012, India Received: 7 April 2017/ Accepted: 17 June 2017/ Published online: 24 July 2017 © Indian Phytopathological Society 2017 ABSTRACT: Histopathological studies of false smut balls on rice spikelet were conducted through SEM and microtomy. Detection and identification of the fungal culture of Ustilaginoidea virens from infected host tissue was done through polymerase chain reaction (PCR) using U. virens specific internal transcribed spacer (ITS) primer. The primer amplified 380bp product. Keywords: False smut balls, microtomy, scanning electron microscopy, Ustilaginoidea virens RESEARCH ARTICLE *Corresponding author: kumaripooja2989@gmail.com False smut of rice is caused by Ustilaginoidea virens, teleomorphic stage is Villosiclava virens. Earlier it was considered as a minor disease of rice (Padwick, 1950; Tanaka et al., 2008) and considered to be a sign of ‘bumper harvest’of the crop. False smut balls on rice panicles are the typical symptom of the disease (Pooja et al., 2015). False smut balls are intertwined with the mycelium of U. virens at early stage and then chlamydospores are formed. The chlamydospores are initially smooth and later become warty at maturity (Kim and Park, 2007; Mathew and Sharma, 2015). Under bright-field light microscopy, the conidia are elliptical and warty on the surface with diameters ranging from 2 to 6 μm. In scanning electron microscopic study the conidia appeared to be globose to irregularly rounded and ornamented with prominent spines. The spines are either pointed or slightly curved at the apex and approximately 200-550 nm long. In transmission electron microscopy, both the spined conidia and hyphae along with lipid globules and vacuoles were seen in the cytoplasm of host cell. The stigma, ovary, and even the young grains of the rice were assumed to be the site of primary infection (Padwick, 1950; Wang, 1992), but subsequently histological observation showed that the pathogen can attack the root of the rice plant at the seedling stage and may colonise symptomlessly in the entire plant (Schroud and TeBeest, 2005; TeBeest, 2010). The fungal life cycle of U. virens and primary infection site on the paddy crop in nature is still very unclear. SEM and TEM study of the ultrathin sections of false smut balls and colonies of U. virens on different media showed that the dense chlamydospores are piled around the inner hyphae. The fungal hyphae and endosperm were not distinguishable. Large stretch of well-developed endosperm was present in the center which was filled with large amount of starch grains. This suggests that the infection of pathogen may also occur in host plant after the flowering and grouting, and the fungal pathogen may be saprophytic in nature. The aim of the present study was to find out the nature of the disease in plants. MATERIALS AND METHODS Spore observation Chlamydospores of U. virens were assessed for their variation using compound and scanning electron microscope. Compound microscope Fungal spores were placed over the cleaned glass-slide having a drop of distilled water using a needle and covered with a cover slip and viewed under the compound microscope under 40X and oil immersion at 100X. The spores were observed for colour and shapes of conidia. The resulting image was then photographed. The cultures were also observed and characters of hyphae and septation were noted. Scanning electron microscopy (SEM) The spores of U. virens were dusted over the specimen stub and sputter coated with palladium (24 nm thick) under vacuum for an hour. The specimens were then observed under Zeiss EVOMA10 SEM in high vacuum. The surface morphology, size, hyphal width and spines of the conidia were measured and the photographs were taken under different magnifications ranging from 2.00-