Abstract:The use of liquid instead of solid culture medium for the micropropagation of plants offers advantages such as better access to medium components and scalability through possible automation of the processes. The objective of this work was to compare a new temporary immersion bioreactor (TIB) to solid medium culture for the micropropagation of a selection of tree species micropropagated for commercial use: Nordmann fir (Abies nordmanniana (Steven) Spach), Eucalyptus (E. grandis x E. urophylla), Downy birch (Betula pubescens Ehrh), and Curly birch (Betula pendula var. carelica). Cultivation of explants in the TIB resulted in a significant increase of multiplication rate and fresh weight of Eucalyptus and B. pendula, but not Betula pubescens. In addition, the fresh weight of embryogenic tissue and the maturation frequency of somatic embryos increased significantly when an embryogenic cell line of A. nordmanniana was cultivated in the TIB compared to solid culture medium. These results demonstrate the potential for scaling up and automating micropropagation by shoot multiplication and somatic embryogenesis in commercial tree species using a temporary immersion bioreactor.
Acetohydroxyacid synthase (AHAS) is the target enzyme for a number of herbicides. A S653N mutation in the AHAS gene results in an increased tolerance to imidazolinone herbicides. We have investigated the use of the mutated gene as selection gene for potato transformation. This resulted in a transformation system with a very high transformation frequency and low rate of escapes. The mutated AHAS gene was introduced into transformed potato together with a beta-glucuronidase (GUS) gene. Selection on 0.5 microM Imazamox yielded GUS expression in 93-100% of regenerated shoots. Furthermore the mutated AHAS gene was used as selection gene for production of high-amylopectin potato lines. The high transformation frequency was verified and potato lines with the desirable starch quality were obtained.
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