Induction of tumors and teratomous shoots and gene transformation of pinus thunbergii and populus koreana x populus nigra var italica by agrobacterium tumefaciens

Choi, W.Y.; Lee, K.J.

Seoul National University Journal of Agricultural Sciences 15(1): 1-6


Accession: 007451951

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Pinus thunbergii and Populus koreana .times. P. nigra var. italica were inoculated with three wild type strains of Agrobacterium tumefaciens (pTiBo542, pTiC58, and Ach5) to test the susceptibility, to induce transformed plants, and to confirm the transformation by opine analysis. P. thunbergii seedlings in vitro (2-month-old) and in vivo (6-month-old) formed tumors in responses to the inoculation with strain pTiBo542 with 10% frequency and pTiC58 and 9-10%. No tumors, however, were observed on seedlings or germinants inoculated with strain Ach5 octopine type. Suspension-cultured cells of Populus koreana .times. P. nigra var. italica were cocultivated with each strain of A. tumefaciens. Microcalli were obtained from plated cells inoculated with strain of pTiBo542 and Ach5. About 5 to 7% of cali continued to growth without phytohormone. Both in vitro leaf slices and intact plants of the poplar formed tumors in response to the strain pTiBo542 with frequency of 77-82%, pTiC58 with 64-79% and Ach5 with 67-71%. Morphological characteristics such as color, surface texture of the tumors were slightly different in each strain. The best combination of growth regulators for teratomous shoot induction was 1.0 mg/l BA and 0.05 mg/l NAA containing 500 mg/l cefotaxime in modified Murashige and Skoog's medium, with induction frequency of 95%. Agropine was detected by electrophoresis in Pinus thunbergii tumors inoculated with pTiBo542, while nopaline was detected in tumors and teratomous shoots of Poulus koreana .times. P. nigra var. italica inoculated with pTiC58.