Transgenic Campanula carpatica plants with reduced ethylene sensitivity

Fertile transgenic Campanula carpatica Jacq. plants with flowers, which had reduced sensitivity to ethylene were obtained by Agrobacterium tumefaciens that mediated transformation. The construct used for transformation contained the etr1-1 gene from Arabidopsis thaliana under control of the flower specific fbp1-promoter from petunia. More than 100 flowering T0 lines were tested for their ethylene sensitivity using 2 microl l(-1) ethylene. The tolerance level to ethylene varied among the lines. While control plants stopped flowering within 3 days of exposure to ethylene, one of the transformed lines flowered for up to 27 days. The presence and the expression pattern of the transgene in various tissues were studied by polymerase chain reaction (PCR) and reverse transcription (RT)-PCR techniques. The expression of etr1-1 was significant in flowers and buds. Transgenic lines did not differ morphologically from control plants. The selected transgenic T0 lines, which were re-established from in vitro cultures showed the same degree of tolerance to exogenous ethylene, confirming the stability of the transgene in in vitro cultures. The rooting ability of the transgenic plants was not affected by the presence of etr1-1. T1 progeny were produced by crossing the transgenic line, which showed the most significant reduction in ethylene sensitivity with a control plant, and the analysis of the T1 plants showed 1:1 segregation in terms of ethylene sensitivity and the presence of the transgene.