Abstract
The PCR based RAPD analysis was used to study genetic relationship among roselle and kenaf accessions. Extraction of genomic DNA samples of nine roselle (Hibiscus sabdariffa L.) and seven kenaf (Hibiscus cannabinus L.) accessions were successfully performed by Cetyl Trimethyl Ammonium Bromide (CTAB) method. RAPD analysis using two chloroplast primers and one mitochondrial primer produced about 62 bands of different sizes. Most of the fragments were polymorphic where 2-3 fragments were monomorphic in each primer. Sixteen accessions were classified into two main groups A and B through dendogram generated using RAPD polymorphism. Five kenaf accessions were included in group A, whereas nine roselle accessions and two kenaf accessions were included in group B. A group was formed with a range of similarity coefficients of 0.5333 to 0.8780 having all roselle accessions. Bengkalis was different from Hibiscus sp but it was close to roselle accessions. Noonsoon was very close to roselle accessions having a range of jaccard similarity coefficients of 0.5000 to 0.6326. It was evident from jaccad similarity coefficient of 0.8780 that the roselle varieties UKMR 1 and UKMR 2 were genetically closer to each other. On the other hand, V36 and Thainung showed the highest similarity coefficient (0.7878) among kenaf accessions in group A. This study has successfully differentiated between two species by using chloroplast and mitochondrial markers which was due to high level of polymorphism. However, the chloroplast primers were more effective than M13 to study the genetic variation between roselle and kenaf.
Highlights
Hibiscus L. is the type genus of the tribeHibisceae of the family Malvaceae (BarssumWaalkes, 1966)
DNA extraction DNA extracted from young leaves of roselle and kenaf by using Cetyl Trimethyl Ammonium Bromide (CTAB) method (Murray and Thompson, 1980) which is a modified version of Doyle and Doyle’s (1990) method
The mixture was treated with 200 μl chloroform: isoamyl alcohol (CIA) (24:1) and shaken gently by inverting the tubes 8-10 times centrifuged at 13000 rpm for 15 min
Summary
Hibiscus L. is the type genus of the tribe. Hibisceae of the family Malvaceae (BarssumWaalkes, 1966). The genus contains about 300 species that grow in tropical and subtropical regions throughout the world (Anderson and Pharis, 2003). It includes both annual and perennial herbaceous plants. Received 11 May 2013; Revised 10 August 2013; Accepted 09 September 2013; Published Online 11 October 2013. M. Aminul Islam Department of Genetics and Plant Breeding, Faculty of Agriculture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh
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