Hindawi Publishing Corporation ISRN Bacteriology Volume 2013, Article ID 562014, 12 pages http://dx.doi.org/10.1155/2013/562014 Research Article Rapid Identification of Polyhydroxyalkanoate Accumulating Members of Bacillales Using Internal Primers for phaC Gene of Bacillus megaterium Pramoda Kumar Nayak, 1 Ajeet Kumar Mohanty, 2 Teja Gaonkar, 1 Ashwani Kumar, 2 Saroj N. Bhosle, 1 and Sandeep Garg 1 1 Department of Microbiology, Goa University, Taleigao Plateau, Goa 403206, India 2 National Institute of Malaria Research, Field Station, DHS Building, Campal, Panaji, Goa 403001, India Correspondence should be addressed to Sandeep Garg; sandeep garg68@yahoo.co.in Received 16 April 2013; Accepted 5 June 2013 Academic Editors: D. A. Bazylinski, G. Nychas, and A. Poli Copyright © 2013 Pramoda Kumar Nayak et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Bacillus megaterium is gaining recognition as an experimental model and biotechnologically important microorganism. Recently, descriptions of new strains of B. megaterium and closely related species isolated from diverse habitats have increased. erefore, its identification requires several tests in combination which is usually time consuming and difficult to do. We propose using the uniqueness of the polyhydroxyalkanoate synthase C gene of B. megaterium in designing primers that amplify the 0.9 kb region of the phaC for its identification. e PCR method was optimized to amplify 0.9kb region of phaC gene. Aſter optimization of the PCR reaction, two methods were investigated in detail. Method I gave an amplification of a single band of 0.9kb only in B. megaterium and was demonstrated by several strains of B. megaterium isolated from different habitats. e use of Method I did not result in the amplification of the phaC gene with other members of Bacillales. e specificity for identification of B. megaterium was confirmed using sequencing of amplicon and RT-PCR. Method II showed multiple banding patterns of nonspecific amplicons among polyhydroxyalkanoate accumulating members of Bacillales unique to the respective species. ese methods are rapid and specific for the identification of polyhydroxyalkanoate accumulating B. megaterium and members of Bacillales. 1. Introduction Bacillus megaterium is a Gram-positive, aerobic, spore form- ing bacterium present in diverse habitats from terrestrial to marine sediments. is “big beast” has been identified as an experimental organism for studies on various cell structures and functions [1, 2]. Its ecological and economical value has been established and reviewed [3–5]. It is only in the past two decades that various new strains of this species with immense potential in industries have been isolated from various ecosystems. e presence of B. megaterium in soil has been implicated in the degradation of herbicides and insecticides [6, 7]. Strains of B. megaterium have been recognised for biological control of root infections due to Rhizoctonia solani [8]. Strains producing biomolecules, such as phytohormone, 2-pentylfuran, pyruvate, oxetanocin, vitamin B12, and polyhydroxyalkanoate (PHA) which are of industrial importance have been investigated [9–12]. is bacterium produces proteins of economical importance, namely, agarase, chitosanase, penicillin G acylase, amylases, glucose dehydrogenase, neutral protease, and the family of P-450 cytochrome monooxygenases [13–15]. Bacillus mega- terium is also amenable for recombinant DNA technology for the improvement of specific enzymes and for protein production [5]. Genomes of biotechnologically important strains of B. megaterium have been sequenced in 2011 [16]. Recently, it has been recognised as a promising candidate among Gram-positive organisms for producing polyhydrox- yalkanoate on an industrial scale because few strains are capable of accumulating the homopolymer and copolymer in the presence of a single carbon source or inexpensive carbon substrates [17–19].