Isoflavone Glycoside Formation in Transformed and Non-Transformed Suspension and Hairy Root Cultures of Lupinus polyphyllus and Lupinus hartwegii

Berlin, Jochen; Fecker, Lothar F.; Rügenhagen, C.; Sator, Christine; Strack, Dieter; Witte, Ludger; Wray, Victor

doi:10.1515/znc-1991-9-1003

Cited by 24 publications

(9 citation statements)

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“…A problem experienced with the inoculation of leaf explants, using either method, was that the bacteria grew very rapidly and frequently engulfed the entire explant in as little as 2 days. This problem of excessive bacterial growth has been noted by other authors [ 1,141. Under such circumstances, it is difficult to imagine any root initiation occurring.…”

Section: Resultssupporting

confidence: 59%

Initiation of and solasodine production in hairy root cultures of Solanum mauritianum Scop.

Drewes¹,

Staden²

1995

Plant Growth Regul

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Initiationand establishment of hairy root cultures from leaf or seedling hypocotyl explants of Solarium mauritianum Stop., using six strains OfAgrobacterium rhizogenes was attempted. Success was only achieved following hypocotyl inoculation with strain LBA 9402. Transformation frequency was very low, with only one instance out of a possible 90 being recorded. Resultant hairy root cultures grew rapidly and could be maintained using a Murashige and Skoog (1962) medium supplemented with 0.1 g L-' myo-inositol and 3% sucrose, either as a solid or liquid culture. Under these conditions, the roots had a solasodine content of 126 pg g-' DW. Lower levels of solasodine and decreased root growth rates were recorded when the medium strength was reduced by half or 3% glucose substituted for the 3% sucrose.Abbreviations: MS -Murashige and Skoog's (1962) medium.

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Section: Resultssupporting

confidence: 59%

Initiation of and solasodine production in hairy root cultures of Solanum mauritianum Scop.

Drewes¹,

Staden²

1995

Plant Growth Regul

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“…Plant DNA for Southern tests was isolated as previously described [2]. For preparation of total RNA for northern blot hybridizations 1-2 g of fresh root tissue, which had been previously blotted dry, were homogenized in a solution containing 4 M guanidinium thiocyanate, 0.5 ~o (w/v) sodium lauroyl sarcosine, 25 mM sodium citrate (pH 7), 0.33% antifoam A (Sigma) and 100 mM 2-mercaptoethanol.…”

Section: Isolation and Analyses Of Dna And Rnamentioning

confidence: 99%

“…Electrophoresis of total RNA (2.5-10/~g) after denaturation with glyoxal and transfer of the RNA to Pall nylon filters were performed as described previously [7]. For hybridizations the Hind III fragment with the ldc expression cassette from pLX154.3 was labelled to a specific activity of 108-109 cpm/#g DNA with c~-[ 32p]-dCTP as described previously [2]. After hybridizations the filters were washed with 0.1°/O SSC, 0.2~o SDS at 56 °C and exposed to Kodak XAR films at -70 °C.…”

Section: Isolation and Analyses Of Dna And Rnamentioning

confidence: 99%

Increased production of cadaverine and anabasine in hairy root cultures of Nicotiana tabacum expressing a bacterial lysine decarboxylase gene

Fecker¹,

Rügenhagen

Berlin

1993

Plant Mol Biol

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Several hairy root cultures of Nicotiana tabacum varieties, carrying two direct repeats of a bacterial lysine decarboxylase (ldc) gene controlled by the cauliflower mosaic virus (CaMV) 35S promoter expressed LDC activity up to 1 pkat/mg protein. Such activity was, for example, sufficient to increase cadaverine levels of the best line SR3/1-K1,2 from ca. 50 micrograms (control cultures) to about 700 micrograms/g dry mass. Some of the overproduced cadaverine of this line was used for the formation of anabasine, as shown by a 3-fold increase of this alkaloid. In transgenic lines with lower LDC activity the changes of cadaverine and anabasine levels were correspondingly lower and sometimes hardly distinguishable from controls. Feeding of lysine to root cultures, even to those with low LDC activity, greatly enhanced cadaverine and anabasine levels, while the amino acid had no or very little effect on controls and LDC-negative lines.

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“…Furthermore, of the limited variety of natural products that have been produced by cultured cells, flavonoid compounds are the least represented (4,9). Among the few examples that have been reported in the literature are isoflavonoids formed in response to biotic or abiotic elicitors (1,3,11) or those produced by cell suspension and hairy root cultures transformed with Agrobacterium tumefaciens and A. rhizogenes, respectively (2).…”

mentioning

confidence: 99%

Biosynthesis of White Lupin Isoflavonoids from [U-¹⁴C]l-Phenylalanine and Their Release into the Culture Medium

Gagnon

Séguin²,

Bleichert³

et al. 1992

Plant Physiol.

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Pulse-labeling experiments of white lupin (Lupinus albus L.) cell cultures with [U-14C1L-phenylalanine for 72 h resulted in the incorporation of the radioactivity into the isoflavone aglucones, glucosides, and prenylated derivatives. Both the aglucones genistein and 2'-hydroxygenistein and their 7-0-glucosides accounted for 85% of the total isoflavonoids identified in the cultured cells and contained 35% of the radioactivity, whereas the prenylated derivatives comprised 15 and 65%, respectively. Almost 20% of the labeled isoflavones of the cellular pool was recovered from the culture medium, 90% of which were monoprenylated and diprenylated derivatives containing 80% of the radioactivity. These results clearly demonstrate the release into the culture medium of a substantial amount of the endogenously synthesized isoflavonoids, especially the prenylated derivatives.With few exceptions, most plant cell cultures fail to produce the secondary metabolites/natural products characteristic of the intact plants from which they were derived (13,17). Furthermore, of the limited variety of natural products that have been produced by cultured cells, flavonoid compounds are the least represented (4, 9). Among the few examples that have been reported in the literature are isoflavonoids formed in response to biotic or abiotic elicitors (1,3,11) or those produced by cell suspension and hairy root cultures transformed with Agrobacterium tumefaciens and A. rhizogenes, respectively (2).Cell cultures of white lupin (Lupinus albus L.) accumulate a complex pattern of isoflavones ( Fig. 1) [3-5, 3a-5a], and 6,3'-diprenyl [6, 6a] The isoflavonoid pattern is quite similar to that of the intact lupin roots (8,10,18,19). However, it was observed that lupin cell cultures release a portion of their metabolites into the culture medium, the latter eventually turning reddishbrown in color. Microscopic examination revealed that this phenomenon was not due to lysis of the cultured cells because the latter remained viable until the end of the culture period. It was considered important, therefore, to investigate the biosynthesis of lupin isoflavones, using "4C-labeled L-phenylalanine as a precursor, and their release into the culture medium. MATERIALS AND METHODS Cell CultureA cell-suspension culture derived from the radicles of white lupin (Lupinus albus L. cv Kievskij Mutant) was maintained on B5 nutrient medium (6) containing 2% sucrose and supplemented with 2,4-D (1 ppm) and kinetin (0.1 ppm). Preparation of Cell and Medium ExtractsAt the end of the metabolic period, the cultured cells were separated from the media by centrifugation at 2000g and washed twice with distilled water. The cells were extracted twice by homogenization with 80% aqueous MeOH4 (5:1, w/ v) at room temperature. The combined extracts were evaporated to an aqueous residue in a rotary evaporator at 300C, and the latter extracted twice with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate before evaporation, and the residue dissolved in 8...

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Isoflavone Glycoside Formation in Transformed and Non-Transformed Suspension and Hairy Root Cultures of Lupinus polyphyllus and Lupinus hartwegii

Cited by 24 publications

References 0 publications

Initiation of and solasodine production in hairy root cultures of Solanum mauritianum Scop.

Initiation of and solasodine production in hairy root cultures of Solanum mauritianum Scop.

Increased production of cadaverine and anabasine in hairy root cultures of Nicotiana tabacum expressing a bacterial lysine decarboxylase gene

Biosynthesis of White Lupin Isoflavonoids from [U-¹⁴C]l-Phenylalanine and Their Release into the Culture Medium

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Isoflavone Glycoside Formation in Transformed and Non-Transformed Suspension and Hairy Root Cultures of Lupinus polyphyllus and Lupinus hartwegii

Cited by 24 publications

References 0 publications

Initiation of and solasodine production in hairy root cultures of Solanum mauritianum Scop.

Initiation of and solasodine production in hairy root cultures of Solanum mauritianum Scop.

Increased production of cadaverine and anabasine in hairy root cultures of Nicotiana tabacum expressing a bacterial lysine decarboxylase gene

Biosynthesis of White Lupin Isoflavonoids from [U-14C]l-Phenylalanine and Their Release into the Culture Medium

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Product

Resources

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Biosynthesis of White Lupin Isoflavonoids from [U-¹⁴C]l-Phenylalanine and Their Release into the Culture Medium