Elimination of sugarcane grassy shoot disease through apical meristem culture A.K. Tiwari a *, S. Tripathi a , M. Lal a , M.L. Sharma a and P. Chiemsombat b a Division of Tissue Culture, UP Council of Sugarcane and Research, Shahjahanpur 242 001, UP, India; b Faculty of Agriculture, Department of Plant Pathology, Kasetsart University, Kamphaeng Campus, Nakhon Pathon 73140, Thailand (Received 9 October 2010; final version received 16 October 2010) Elimination of sugarcane grassy shoot disease (SGSD) through apical meristem culture technique for producing clean planting material of sugarcane has been attempted in the present study. The results showed that meristems length of 2 and 3 mm were free from the SGSD pathogen at higher frequency than larger meristem length of 4 mm. However, the frequency of survival of explants during initiation of shoot cultures was higher in larger meristems (60%) in comparison to smaller ones (40%). The micropropagated plantlets raised from meristem culture were confirmed for disease-free by nested polymerase chain reaction (PCR) analysis at monthly interval up to 6 months. This is the first report on the elimination of SGSD phytoplasma through meristem culture in India. Keywords: sugarcane; grassy shoot disease; elimination; meristem culture; phytoplasma Introduction Sugarcane (Saccharum officinarum), an important agricultural crop of India, is affected by several pathogens such as viruses, phytoplasma, fungi and bacteria that are responsible for reducing the yield and quality of the crop. The occurrence of sugarcane grassy shoot disease (SGSD) has reached to the alarming level in India (Vishwanathan 2000). The disease is associated with sugarcane grassy shoot phytoplamsa, which affects the phloem of infected plants and is classified as a member of the 16Sr XI-B group (Lee et al. 1998). The disease causes severe loss to millable canes in plant crops and the severity increases multifold in ratoon crops. The occurrence of SGSD has been reported from Bangladesh, India, Malaysia, Nepal and Pakistan (Vishwanathan 2001; Rao et al. 2008). According to heavy yield loss caused by SGSD, the supply of pathogen-free planting material is pivotal to sustainable crop production. Phytoplasmal diseases are very problematic specially in vegetatively propagated crops in which they are transmitted from generation to generation through the planting materials (Wang et al. 2008; Tiwari et al. 2010). Earlier efforts on chemotherapy and thermotherapy alone or in combination with in vitro tissue culture technique (Chatenet et al. 2001; Balamuralikrishnan et al. 2002; Parmessur et al. 2002; Tiwari et al. 2008; Mishra et al. *Corresponding author. Email: ajay_biotech2005@rediffmail.com Archives of Phytopathology and Plant Protection Vol. 44, No. 20, December 2011, 1942–1948 ISSN 0323-5408 print/ISSN 1477-2906 online Ó 2011 Taylor & Francis http://dx.doi.org/10.1080/03235408.2010.544446 http://www.tandfonline.com