Improved methodology for the isolation of false smut pathogen Ustilaginoidea virens of rice
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Improved methodology for the isolation of false smut pathogen Ustilaginoidea virens of rice B. M. Bashyal1 · Pooja Parmar1 · Najam Waris Zaidi2 · Sunil Kumar Sunani3 · G. Prakash1 · Rashmi Aggarwal1 Received: 28 May 2020 / Revised: 11 September 2020 / Accepted: 17 September 2020 © Indian Phytopathological Society 2020
Abstract Rice false smut is an emerging disease of rice caused by Ustilaginoidea virens (Cooke) Takashi. Along with yield loss, the pathogen affects the quality of the grains and produces mycotoxins also. Isolation of the pathogen is difficult due to its slow growing nature which often leads to the contamination. Hence, an improved methodology for the isolation was developed. For this, first the infected rice smut balls were taken and sterilized for 2 min in 1% (v/v) sodium hypochloride solution followed by washing three times (1 min each) with sterilized distilled water. Then, the smut balls were halved using a sterilized scalpel and spores from the inner surface of the ball were dusted onto the Petri plates containing ampicillin added potato sucrose agar (PSA). The plates were incubated at 27 °C ± 1 °C for 7 days after which white fungal colonies appeared were transferred in the PSA plates. The pathogen U. virens was further confirmed morphologically and through internal transcribed spacer (ITS) region amplification with 98–100% sequence homology. The developed methodology is comparatively faster with 80–85% success and will be helpful for the pathogen characterization, and understanding the biology of the pathogen. Keywords False smut · Ustilaginoidea virens · Rice · Isolation method Rice is a major staple food worldwide. However, the crop productivity is affected by various biotic and abiotic stresses. Rice false smut caused by Ustilaginoidea virens is an emerging disease of rice recorded with an incidence upto 50% in major rice growing countries worldwide (Nessa et al. 2015). The disease was initially thought to be an indicator of good harvest. However, increasing instances of false smut are attributed to modern agriculture practices such as use of chemical pesticides, high yielding varieties and increase in global temperature at booting stage (Teng 1990). Besides these the changing global and regional climatic conditions are aggravating the problem (Rush et al. 2000; Guo et al. 2012). The disease is characterized by replacement of rice grains into yellow false smut balls which later on converted to dark green. Additionally, the ustiloxins produced by the * B. M. Bashyal [email protected] 1
Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi 110012, India
2
International Rice Research Institute, New Delhi 110008, India
3
Mizoram Centre, ICAR Research Complex for NEH Region, Kolasib, Mizoram 796081, India
fungi are compromising the quality of rice grains and are inevitably added to the harvest (Lu et al. 2010; Zhou et al. 2012). In spite of increasing importance of the disease the studies related to the disease occur
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