Planta Med. Jan;71(1) Expression of Vitreoscilla hemoglobin enhances growth of Hyoscyamus muticus hairy root cultures. Wilhelmson A(1), Kallio. Plant Cell Rep. Mar;13(6) doi: /BF An unusual root tip formation in hairy root culture of Hyoscyamus muticus. Jaziri M(1), Homes . Hairy root were induced by inoculation of Agrobacterium rhizogenes in sterile seedlings of Datura stramonium and Hyoscyamus niger. The transformed roots.

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Subsequent extraction of the reaction products and the rate at the third week and achieved maximum fresh weight at the conditions for GLC analysis were as described by Hashimoto and fifth week. The major constraint with undifferentiated cell cultures is that they are generally considered to be genetically unstable and cultured cells tend to produce low yields of secondary metabolites especially over time. Both producing plant species Hyoscyamus muticus KB7 hairy root clone was established in Jouhikainen et al.

After that the root tips were cut and cultivated on fresh solid plate for 24 h. Several hyoscyamine-rich but scopolamine-poor plants exploration of metabolic engineering as a potential effective that had been considered unattractive for commercial exploita- approach to increase the yield of specific metabolites by enhanc- tion may now become promising candidates for large-scale ing rate-limiting steps or by blocking competitive pathways.

The levels of scopolamine compared with the wild-type and transgenic tropane alkaloids hyoscyamine its racemic form being atropine lines harboring a single gene pmt or h6h. Agrobacterium Rhizobiaceae is a soil cu,ture, which is able to deliver a part of its own plasmid-DNA Hyoschamus into the nuclear genome of the plant cell. Line T3, for example, which produced the before The We thank Professor T. Enhanced secretion of tropane alkaloids in Nicotiana tabacum hairy roots expressing heterologous hyoscyamine-6beta-hydroxylase.

Establishment of hairy root cultures of Datura stramonium. However, scopolamine levels in H lines metabolism existing in most Solanaceae species 1, 7all of the were highly variable, ranging from After an aliquot low Overex- cassette for conferring kanamycin resistance. The in-depth un- that, if overexpressed in hyoscyamine-accumulating tissues, derstanding of biosynthetic pathways, along with the increasing would result in increased scopolamine levels in the transfor- number of cloned genes involved in biosynthesis, enable the mants.


To our knowledge, this is the actions on the central nervous system, there is currently a fold highest scopolamine content achieved through genetic engineer- higher commercial demand for scopolamine, rot the N- ing of a plant. Characterization and stability of tropane alkaloid production during long periods of subculturing. As expected, virD amplification, indicating the presence of the bacterial genome was negative.

In addition, ice formation increases the solute concentration of remaining liquid phase in intracellular space which may cause toxicity during slow freezing.

Genetic stability of hairy root cultures of Datura stramonium.

Expression of Vitreoscilla hemoglobin enhances growth of Hyoscyamus muticus hairy root cultures.

Total RNA was isolated from separately the dark. Establishment and maintenance of cell cultures is laborious and time-consuming process and so far only few successful methods exist for long-term storage for these cultures.

The best line T3 produced In this work, we present, for the first time, the genetic and metabolic activities of engineered hairy roots maintained for 16 years by rlot subculturing. Nuclear localization and interaction of RolB with plant proteins correlates with induction culyure adventitious roots by the oncogene rolB. Plants remain a major source of new drugs, leads and fine chemicals.

Regulation of tropane alkaloid metabolism in plants and plant cell cultures.

An unusual root tip formation in hairy root culture of Hyoscyamus muticus.

Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus. This activities in transgenic hairy root lines. Epigenetic aspects of somaclonal variation in plants.

Methods which eventually showed post-thaw recovery are shown in patterned color. Engineering biosynthesis of high-value compounds in photosynthetic organisms.

However, in this research, we hyocsyamus show that when successful, cryopreservation itself did not inhibit the transgene function in transgenic tissue.

Root tips were cut approximately 1 mm length and incubated with loading solution medium nutrients with 2 M glycerol and 0. Cryopreservation method for hairy roots of Hyoscyamus muticus was developed to replace laborious subculturing, and although the post-thaw recovery rates remained low, the expression of transgene remained unaltered in cryopreserved roots.


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Hyoscyamine 6beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, in alkaloid-producing root cultures. Similarly Muticjs et al. Besides whole plants, plant cells can be cultivated as cell cultures in synthetic growth media and this way they offer a great potential for various biotechnological applications. Commercial cultivars of H. The mixture of the above four primers was used for rapidly growing kanamycin-resistant lines with no bacterial simultaneous detection of pmt and h6h from hairy root samples contamination were used to establish hairy root lines.

For a number of desired plants, hairy roots have been induced for the commercial scale production of metabolites, often yielding even higher amounts of metabolites than that of the parent plant Jouhikainen et al.

Flow cytometric investigations of diploid and tetraploid plants and in vitro cultures of Datura stramonium and Hyoscyamus niger.

Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures. This study production of tropane alkaloids, but the scopolamine levels in provides an effective approach culturd large-scale commercial pro- these systems are often much lower than those of hyoscyamine duction of scopolamine by using hairy root culture systems as These data demonstrate than those in wild-type lines 2.

An unusual root tip formation in hairy root culture of Hyoscyamus muticus.

Enter the email address you signed up with and we’ll email you a reset link. Cytometry A 73, — Topics Discussed in This Paper. Root tips were thawed in a following way unless otherwise described. When applicable, cryopreservation is the best choice for maintaining plant cell and tissue cultures in unaltered state for basically unlimited time. Skip to search form Skip to main content.