배발생 캘러스를 이용한 아그로박테리움 매개형질전환 장미 식물체 획득

Acquirement of transgenic rose plants from embryogenic calluses via Agrobacterium tumefaciens

The process to acquire intron-GUS geneexpressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. `Sweet Yellow` using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D=0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with 250 mg·L-1 cefotaxim at 4℃ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim 250 mg·L-1 and ppt 2 mg·L-1. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim 250 mg·L-1 and ppt 2 mg·L-1, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim 250 mg·L-1 and ppt 2 mg·L-1 to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim 250 mg·L-1. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.