Assessment of genetic diversity among Asparagus species using randomly amplified polymorphic DNA markers Bhuvaneshwaragouda R Patil • Mahantesh Talikoti • Sanjeev K Deshpande • CG Patil • Malagouda Patil Received: 3 March 2012 / Accepted: 30 April 2012 © CISI 2011 Bhuvaneshwaragouda R Patil ( ) ) • Mahantesh Talikoti • Sanjeev K Deshpande • Malagouda Patil Department of Genetics and Plant Breeding University of Agricultural Sciences, Dharwad, Karnataka CG Patil Karnataka College, Dharwad Abstract Thirty five random decamer primers used in the present study amplified the genomic DNA of seven Asparagus species. A total of 554 bands were observed. Highest pair-wise similarities (0.72) were observed between Asparagus umbellatus and Asparagus racemosus. The lowest similarity (0.31) was found between Asparagus densiflorus ‘Sprengeri’ and Asparagus umbellatus ; Asparagus Officinalis; Asparagus racemosus. The random decamers successfully discriminated seven Asparagus species. Keywords: RAPD, molecular markers, principal component analysis Introduction Asparagus is an important vegetable crop grown in a wide diversity of environments ranging from cool temperate zones (Germany, Netherlands) to deserts (Peru), Mediterranean climates (California, Spain) and tropical areas (Philippines, Taiwan). It has various uses; important ones apart from vegetable are medicinal and ornamental purposes. This “Royal Vegetable” belongs to lily family and consists of 300 species [Madhavan et al., 2010]. The plant exhibits bell shaped, greenish white to yellowish sexually dimorphic flowers occurring in different parts of the same plant.It is a perennial herb to shrub with an underground tuberous stem or root possessing woody stem and leaves reduced to cladodes, growing straight or as a climber. Asparagus seems to be native of Europe, Northern Africa and Western Asia; and now widely grown in both the hemispheres. The exact origin of this vegetable is not known because many wild species are found growing throughout Europe and Asia. However, the most likely location is around the Mediterranean sea where cultivation of this plant has been practiced for over 2000 years, first by the Greeks and then by the Romans (200 B.C.) . It has been considered a delicacy since ancient times; somehow the popularity waned during middle Ages and again became popular in the modern times. Today, Peru is the leading producer of asparagus followed by USA and Spain. Limited information with respect to the genetic diversity among the species of the genus Asparagus is available. Present study was undertaken to assess the genetic diversity of Asparagus species collected from the various geographical locations of Karnataka, India. Materials and methods The seven Asparagus plant species (Table 1) used in the present study were collected from various geographical locations of Karnataka and are maintained in Botanical garden.One set of identified and vouchered specimens was deposited in Herbarium, Department of Botany, Karnataka Science College, Dharwad. Genomic DNA of all the seven Asparagus species was isolated according to modified CTAB method (Doyle et al., 1990) with minor modifications. The yield of DNA per gram of tissue isolated was measured using an Ultra Violet spectrophotometer at 260/280 nm. The purity of DNA was determined by estimating the ratio of absorbance at 260/280 nm. DNA concentration and purity was also reconfirmed by running the samples on 1 per cent agarose gel. The pure DNA yielded a single crisp band. Amplifications were carried out using a DNA thermal cycler (Mastercycler gradient, Eppendorf). Each 20 μl reaction mixture contained about 50 ng of template DNA, 10X PCR Buffer (Tris with 15mM MgCl 2 ) [Bangalore Genei, India], Crop Improv (2012) 39(1): 65-69 Available online at www.indianjournals.com