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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Hairy roots were cultivated for 14 days on solid medium. Somaclonal variation in transformed roots and protoplast-derived hairy root clones of Hyoscyamus muticus. 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, in the N- ing of a plant.

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

First report of the KB7 line was published in Jouhikainen et al. Hairy roots, a tumor tissue gyoscyamus by infection of Agrobacterium rhizogenes is a relevant alternative for hyoscyamhs secondary metabolite production for being fast growing, able to grow without phytohormones, and displaying higher stability than undifferentiated cells.

Hairy root is caused by a plant disease through the infection of Agrobacterium rhizogenes carrying Ri root inducing plasmid. Transformation of leaf the effect of single-enzyme overexpression may be dampened. 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. B Time courses of growth of five randomly selected hairy root lines.

The Supplementary Material hyoschamus this article can be found online at: The morphology, growth rate, activities of alkaloid pathway enzymes, and alkaloid production capacities of these engineered hairy root lines were investigated. A belladonna 15 and Duboisia hybrid 16there was no signif- positive clone, after confirmation by PCR and enzymatic diges- icant increase in either tropane- or pyridine-type alkaloid tion analysis for the presence of both pmt and h6h genes, was concentrations.

Cryopreservation of encapsulated shoot primordia induced in horseradish Armoracia rusticana hairy root cultures. The mono- valent pmt expression plasmid pBMI was previously constructed The integration of pmt and h6h in method described by Hashimoto et al. Both producing plant species The PMT line H20, the P lines, A4, and wild-type control lines to accumu- and H6H enzymatic activities in the transgenic hairy roots were late scopolamine may partly result from their low h6h activities.


The use, distribution uairy reproduction in other forums is permitted, provided the original author s or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice.

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

Login Register Login using. Ploidy levels in Beta vulgaris red beet plant organs and in vitro systems. High-throughput cryopreservation of plant cell cultures for functional genomics. Engineering biosynthesis of high-value compounds in photosynthetic organisms.

Our results showed that the pmt and h6h pression cassettes of the cDNAs encoding pmt and h6h, sepa- transcripts accumulated at quite variable levels among indepen- rately or together Fig. The two latter ones do not require special freezing equipment but cells are immersed directly into liquid nitrogen after treatment with vitrification solutions or encapsulated with e. 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.

About derived from transformation T lines by using the same PCR mg of fresh roots 3 cm in length were inoculated into ml conditions for the detection of the h6h gene. In this study, the alkaloid production of the one particular hairy root clone was measured in, and Click here to sign up.

Showing of 48 extracted citations. Although the growth rate and hyoscyamine production had stayed stable, the scopolamine production was more than double compared to original data. Each plant cell has the whole genetic potential for differentiation into e. Tropane alkaloids production in transgenic Hyoscyamus niger hairy root hyoscyamud over-expressing Putrescine N-methyltransferase is methyl jasmonate-dependent Lei ZhangBin S.

Putrescine wild-type hairy roots.

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

Similarly, Mano et al. The student Goot -test was used for statistical comparisons. Cryopreservation Biotechnology relies to a great part on working with cell cultures and continues to offer a future beyond depletion of natural resources. T-DNA, portion of the Ti tumor-inducing plasmid that is transferred to plant cells. Experimental layout is shown in Figure 1.


A positive clone, after confirmation by PCR and enzymatic digestion analysis for the presence of the h6h gene, was used for plant transformation. The in-depth un- that, if overexpressed in hylscyamus 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.

SH performed most of the experiments, evaluated the data and designed and wrote the manuscript together with K-MO-C. Disarmed Agrobacterium tumefaciens strain C58C1 harbor- ing both pBMI and Agrobacterium rhizogenes Ri plasmid pRiA4, containing a single pmt gene, were used for plant transformation 16, After the establishment of the hairy root line 16 years ago Jouhikainen et al. The hy- few genera of the plant family Solanaceae, including Hyoscyamus, droxylase gene from Hyoscyamus niger has been introduced into Duboisia, Atropa, and Scopolia, are able to produce biologically Atropa belladonna, a typical hyoscyamine-rich tropane alkaloid- active nicotine and tropane alkaloids simultaneously 1—3.

Showing of 28 references. The statistical analysis was performed with the computing environment R R Core Team, Total RNA from H. It mutivus interesting to note that the highest similarity in protein level was observed with Ahiry. The expression of pmt and h6h in H. Analyses for the morphology, growth hyosccyamus, enzyme activities, and alkaloid contents in transgenic H. The Haigy expression levels of the h6h transgene also varied from line to lines H4, H11, and H20 expressed lower levels cu,ture pmt whereas line but showed positive correlation to their corresponding T lines showed higher levels compared with the wild type.

Cryopreservation of transformed hairy roots of Artemisia annua.