Studies of population genetics on four mangrove tree species in Okinawa, Japan

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In this studies, attempts have been made to analyze genetical variation of four mangrove tree species, Bruguiera gymnorrhiza (Rhizophoraceae). Kandelia candel (Rhizophoraceae), Rhizophora stylosa (Rhizophoraceae) and Sonneratia alba (Sonneratiaceac), distributed in Okinawa, Japan, and adding one species, Lumnitzera racemosa (Combretaceae) for DNA analysis. Leaves as samples were collected from trees of four species, and stocked in a deep-freezer before experiment. Polyacrylamide gel electrophoresis (PAGE) of a vertical slab-format method was used for analyzing isozyme loci of four mangroves. After the electrophoresis, 23 enzyme species were stained. Fine zymogrames were obtained five enzyme species for Rhizophora stylosa and five enzyme species for Bruguiera gymnorrhiza. Polymorphism was found only peroxidase (POD) for Bruguiera gymnorrhiza, but diaphorase (DIA), esterase (EST) and glucokinase GR) for Sonneratia alba.DNA was extracted from leaves of Rhizophora stylosa. Both CTAB (Cetyl Trimethyl Ammmonia Bromide) and Ice Cold Buffer methods were applied for DNA extraction. The Ice Cold Buffer method was better than the CTAB method for DNA extraction. To obtain mitochondrial DNA fragments, 18 restriction enzyme species were applied. For amplification of chloroplast DNA, 23 primers were used. Mitokondorial and genom DNA fragments were amplified and analyzed. Since genetical variations were small in studied mangrove species collected in Okinawa, primers applied were not so efficiently worked. However, we could obtain evidence that micro-satellites are one of efficient ways for gentical variation analysis of Rhizophora stylosa in Ryukyu islands.