RAPD marker assisted evaluation of chloroplast DNA variation in twelve hosta taxa

Hostas are major ornamental plants for perennial shady nurseries. They have striking foliage with attractive flowers, and their leaves are sometimes used as vegetables in Japan and Korea. The objective of this study was to estimate the genetic identity and genetic distance of twelve hosta taxa and to build a UPGMA cluster for them using Random Amplified Polymorphic DNA (RAPD). Rhizomes of hostas were collected from Shikoku Island, Japan. Newly emerged unfurled hosta leaves were disrupted with Nuclei Lysis solution and then purified the genomic cpDNA. The cpDNA was dehydrated as a final point for extraction. We amplified the cpDNA of twelve taxa by means of 20 RAPD 10mer markers (Kit A: OPA-01 to OPA-20). The amplified DNA was run in 0.8% agarose gel using 1000 kb DNA ladder. Out of 20 RAPD markers, the genomic cpDNA was successfully amplified for fourteen. Fourteen markers generated a total of 102 fragments. We found maximum genetic distance (0.655) and minimum genetic identity (0.520) between H. sieboldiana H. longissima taxa and between H. sieboldii H. longissima. Estimated minimum genetic distance (0.071) and maximum genetic identity (0.931) were found between H. kikutii var. polyneuron H. longipes var. caduca. The UPGMA dendrogram revealed the relationship among the 12 hostas. The results confirmed our hypothesis of large genetic variability among the taxa, which distinguishes the genotypes clearly. The RAPD 10mer markers were found useful in application to the evaluation of the genetic variability and diversity within taxa.