BEPLS Vol 2 [8] July 2013 78 | P age © AELS, India Bulletin of Environment, Pharmacology and Life Sciences Bull. Env. Pharmacol. Life Sci., Vol 2 (8) July 2013: 78-83 ©2013 Academy for Environment and Life Sciences, India Online ISSN 2277-1808 Journal’s URL:http://www.bepls.com ORIGINAL ARTICLE Evaluation of Genetic Diversity in Bamboo through DNA Marker and Study of Association with Morphological Traits A.Shalini* Rakesh K Meena, Shubhankar Tarafdar and Sanket Thakur Aditya Biotech Lab & Research Pvt. Ltd, Raipur, Chhattisgarh-492001 *Correspondence author: Email- shalini.tech65@gmail.com ABSTRACT The taxonomic studies of the bamboos are based on morphological and growth habit, which can cause problems in identification of genetic relatedness. Identification of genetic relationships in 10 species of bamboo was investigated using random amplified polymorphic DNA (RAPD) technique and their association with morphological characters was studied. Analysis started by using twenty five markers (Twenty one RAPD and four ISSR) that allowed us to distinguish 10 species. Out of twenty five, ten primers were used for identification and for establishing a profiling system to estimate genetic diversity. A total of three hundred ninety alleles were amplified out of this hundred distinct polymorphic DNA fragments (bands), ranging from 1.0–0.2 kb were amplified by 10 selected primers. On the basis of morphological traits, Bambusa tulda and Bambusa balcoa were very similar to each other for all four characters. While Guadia angustifolia showed very different types of character in all four traits but it showed highest number of new culms as compared to others. The genetic similarity analysis was conducted based on presence or absence of bands, which revealed a wide range of variability among the species. Cluster analysis clearly showed two major clusters belonging to 10 species of bamboo. Major Cluster1 was further subdivided into three minor clusters. The species of Bambusa tulda and Bambusa balcoa were the most closely related and formed the third minor cluster along with Bambusa nutans and Dendrocalamus strictus. The variety Guadia angustifolia was very distinct and showed as an out group in the dendrogram that was single species in cluster B. Thus, the morphological characterization was confirmed by the genetic diversity analysis. Key words: Bamboo, RAPD, ISSR, Morphological traits, Genetic diversity, Association Received 21/05/2013 Accepted 30/06/2013 © Academy for Environment and Life Sciences, India INTRODUCTION India has the second largest reserve of bamboo populations in the world. The fibres of bamboo are mainly used in the pulp, paper and charcoal industries, while the culms have several other uses as ‘poor man’s timber’. The total annual bamboo demand in India has been estimated to be approx. 5 million tonnes, of which about 3-5 million tonnes are required by the paper and pulp industry alone [1]. Bambusa balcoa is one of the strongest species and is thus preferred for construction purposes; it is also used for paper pulp production [2]. On the other hand, B. tulda is sturdy, tall and one of the five quick growing (70 cm d_1) species of bamboo [3]. The species is suitable for the production of quality paper due to its long fibres [4] and also for producing furniture. Among the anatomical characteristics of bamboo culm, fibre length is important for technical evaluation, especially in the pulp and fibre-based industries. Fibre wall thickness of the culm predetermines the pulping characteristics, paper quality, permeability and strength relationships [5]. Bambusa balcooa and B. tulda are two abundant tropical species that are recognized as priority bamboo species by the FAO (www.unep-wcmc.org, ww.inbar.int) amongst eighteen other bamboo species found globally. Basic knowledge of the biology and genetics of bamboo is severely lacking. This is a direct result of the unusual life cycle of bamboo. Among bamboo species, the vegetative growth phase varies from 1 year to as long as 120 years, and some species have never been known to flower [6]. Identification of sterile plants is therefore problematic as taxonomic studies of bamboos have traditionally depended heavily on inflorescence and floral morphology because: (1) vegetative characters are often environmentally influenced, which makes them less constant for systematic purposes [7]; (2) characters that delimit species may be more subtle and not available for study; and (3) bamboo clones found in Asia are selected for economic value and are widely distributed without proper identification at the species level. The application of modern molecular techniques is therefore of great assistance in species identification. PCR-based genetic markers are now well documented for species/cultivar identification.[8-10] A number B B B E E E P P P L L L S S S