Pengujian Stabilitas Genetik Plantlet Citrumelo Hasil tTCL dari Kultur In Vitro Dengan Menggunakan Teknik Sekuen Berulang

Farida Yulianti, Hidayatul Arisah, Dita Agisimanto


ABSTRACT. Assessment of genetic stability of long-term cultivation of plantlet derived tTCL citrumelo using repetitive sequence primers. (Pengujian Stabilitas Genetik Plantlet Citrumelo Hasil TTCL dari Kultur Jangka Panjang Menggunakan Sekuen Berulang). Farida Yulianti, Hidayatul Arisah, dan Dita Agisimanto. Regeneration of plantlet from organogenesis of stem transverse thin cell layer (tTCL) was achieved for citrumelo, a valuable rootstock. A 4 repetitive based sequences retrotransposon and inter simple sequence repeat (ISSR) marker assays were used to evaluate genetic stability of a group of  22 monthsold in vitro plantlets and to confirm the most reliable method for true-to-type propagation of citrumelo developed so far. Leaves of plantlets were selected and isolated in bulk. Groups of DNA in bulk segregant analysis (BSA) were amplified and separated using agarose gel. Vitroplants that morphologically similar have been effectively distinguished by a selected primer INT- retrotransposon that detect an deletion band at 550 bp on a line a group of sample. Retrotransposon is abundance through the genome and its activation induced by stress condition. Tissue culture condition was reported potential to induce retrotransposon activation. Thegenetic variation of 2.6% was acceptable for the culture that produced from long-term.  Plantlets used in this study were selected from population induced from early 2014, and employed for studying media as well as environment factors for efficiently organogenesis of citrumelo in period of 2014-2015. However, additional study is on going for evaluating genetic variability from a cycle plantlet production through tTCL of stems. This combination protocol of organogenesis and PCR based markers detection would be powerful tools for mass propagation of high quality seedling derived tissue culture for breeding or cultivation programs. 


Citrumelo, organogenesis, tTCL, genetic variability, retrotransposon, ISSR

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