In Vitro Regeneration of Lycium chinense Miller and Detection of Silent Somaclones with RAPD Polymorphisms

  • Ahn, In-Suk (Department of Forestry, North Carolina University) ;
  • Park, Young-Goo (Department of Forest, Kyungpook National University) ;
  • Shin, Dong-Ill (Department of plant genetic engineering, Daegu Catholic University) ;
  • Sul, Ill-Whan (Department of Biotechnology. Daegu University of Foreign Studies)
  • 발행 : 2004.09.01

초록

An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

키워드

참고문헌

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