Among several naturally occurring environmental factors, temperature is considered to play a predominant role in controlling proper growth and flowering in geophytes. Most of them require a “warm-cold-warm” sequence to complete their annual cycle. The temperature optima for flower meristem induction and the early stages of floral organogenesis vary between nine and 25 °C, followed, in the autumn, by a several-week period of lower temperature (4–9 °C), which enables stem elongation and anthesis. The absence of low temperature treatment leads to slow shoot growth in spring and severe flowering disorders. Numerous studies have shown that the effects of the temperature surrounding the underground organs during the autumn-winter period can lead to important physiological changes in plants, but the mechanism that underlies the relationship between cold treatment and growth is still unclear. In this mini-review, we describe experimental data concerning the temperature requirements for flower initiation and development, shoot elongation, aboveground growth and anthesis in bulbous plants. The physiological processes that occur during autumn-winter periods in bulbs (water status, hormonal balance, respiration, carbohydrate mobilization) and how these changes might provoke disorders in stem elongation and flowering are examined. A model describing the relationship between the cold requirement, auxin and gibberellin interactions and the growth response is proposed.
A fully contained and efficient heterologous protein production system was designed using Brassica rapa rapa (turnip) hairy roots. Two expression cassettes containing a cauliflower mosaic virus (CaMV) 35S promoter with a duplicated enhancer region, an Arabidopsis thaliana sequence encoding a signal peptide and the CaMV polyadenylation signal were constructed. One cassette was used to express the green fluorescent protein (GFP)-encoding gene in hairy roots grown in flasks. A stable and fast-growing hairy root line secreted GFP at >120 mg/l culture medium. GFP represented 60 % of the total soluble proteins in the culture medium. Turnip hairy roots retained sustainable growth and stable GFP production over 3 years. These results were superior to those obtained using tobacco hairy roots.
The Brassica rapa hairy root based expression platform, a turnip hairy root based expression system, is able to produce human complex glycoproteins such as the alpha-L-iduronidase (IDUA) with an activity similar to the one produced by Chinese Hamster Ovary (CHO) cells. In this article, a particular attention has been paid to the N- and O-glycosylation that characterize the alpha-L-iduronidase produced using this hairy root based system. This analysis showed that the recombinant protein is characterized by highly homogeneous post translational profiles enabling a strong batch to batch reproducibility. Indeed, on each of the 6 N-glycosylation sites of the IDUA, a single N-glycan composed of a core Man GlcNAc carrying one beta(1,2)-xylose and one alpha(1,3)-fucose epitope (M3XFGN2) was identified, highlighting the high homogeneity of the production system. Hydroxylation of proline residues and arabinosylation were identified during O-glycosylation analysis, still with a remarkable reproducibility. This platform is thus positioned as an effective and consistent expression system for the production of human complex therapeutic proteins.
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