Dwarf genotypes of the economically important flowering potted plant Kalanchoe blossfeldiana were developed by molecular breeding. Root inducing (Ri)-lines were regenerated by applying CPPU to the hairy roots, which were produced by inoculating leaf explants with a wild-type Agrobacterium rhizogenes strain ATCC15834. Amplification by polymerase chain reaction (PCR) and Southern blot analysis confirmed the presence of T-DNA in the Ri-lines. Six Ri-lines were characterised in a greenhouse trial revealing that several morphological traits changed with respect to ornamental value such as plant height, number of lateral shoots, leaf size, leaf number, flower size and number of flowers. The Ri-lines differed in their degree of Ri-phenotype, and the internodes of the Ri-lines were clearly shorter, giving a compact growth habit compared to control plants. Time to anthesis was the same in Ri-line 331 as in control plants and delayed by only 3 days in Ri-line 306 as compared to control plants. A compact plant without delayed flowering can be assumed to be valuable for further breeding.
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.
Some factors influencing in vitro cultures of potted Hibiscus rosa-sinensis L. using nodal cuttings were investigated. A protocol using a modified MS medium helped to overcome chlorosis, shoot tip necrosis (STN) and leaf drop. These disorders have been caused by mineral imbalance associated with calcium and iron deficiency. STN and leaf drop were overcome by increasing calcium level from 3 mM (MS standard concentration) to 9 mM, and this increase, in addition, enhanced shoot dry weight and shoot extension. The chlorophyll content and leaf area increased by increasing the iron concentration 3-fold from 98 lM to 295 lM. Furthermore, substituting Fe-EDTA with Fe-EDDHA resulted in an increase in chlorophyll content, leaf area and shoot extension. The most suitable multiplication medium for H. rosa-sinensis L. was demonstrated to be a modified MS medium containing 2.2 lM BAP and increased concentrations of calcium at 9 mM and iron at 295 lM provided as Fe-EDDHA. The shoots were rooted in half-strength modified MS medium containing 2.7 lM NAA. Acclimatization was successful with all shoots with or without roots.
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