Hairy Root Cultures of Peganum harmala II. Characterization of Cell Lines and Effect of Culture Conditions on the Accumulation of ß-Carboline Alkaloids and Serotonin

1992 ◽  
Vol 47 (3-4) ◽  
pp. 222-230 ◽  
Author(s):  
J. Berlin ◽  
I. N. Kuzovkina ◽  
C. Rügenhagen ◽  
L. Fecker ◽  
U. Commandeur ◽  
...  
Keyword(s):  
Hairy Root ◽  
Dry Mass ◽  
Culture Conditions ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Peganum Harmala ◽  
Ammonium Ions ◽  
Negative Effect ◽  
Type Strains

Abstract Hairy root cultures of Peganum harmala established by genetic transformation with Agrobacterium rhizogenes wild type strains A4, 15834 or carrying the binary vector pLTCgus 1, respectively, were analyzed with respect to their tryptophan-derived metabolites. Beside the previously detected β-carboline alkaloids harmine, harmol, harmaline and harmalol all hairy root cultures were found to contain the β-carboline glucoside ruine and serotonin. The appearance of the root cultures and the levels of β-carboline alkaloids were greatly affected by the nitrogen source of the medium. In the presence of higher levels of ammonium ions (as in MS medium) the roots were short and thick, while in B50 medium (low ammonium) they were thin and long. Ammonium ions had a negative effect on the levels of β-carbolines. The best root cultures accumulated on B50 1.5-2% β-carbolines on a dry mass basis and in the presence of 6% sucrose ca. 10 mg β-carbolines/L x d were formed. In the presence of phytohorm ones root cultures were converted into low producing suspension cultures, from which the highly productive roots were reinduced on phytohormone-free medium.

Production of β-Carboline Alkaloids in Transformed Root Cultures of Peganum harmala L.

1990 ◽  
Vol 45 (6) ◽  
pp. 727-728 ◽  
Author(s):  
I. N. Kuzovkina ◽  
A. Gohar ◽  
I. E. Alterman
Keyword(s):  
Genetic Transformation ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Dry Mass ◽  
Suspension Cultures ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Peganum Harmala ◽  
Transformed Root Cultures

Abstract Peganum harmala, Hairy Root Cultures, β-Carboline Alkaloids Hairy root cultures of Peganum harmala were established by genetic transformation with Agrobacterium rhizogenes strain A4. In contrast to suspension cultures the root cultures contained high levels of β-carboline alka­ loids (1 -1.5% of dry mass).

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

1991 ◽  
Vol 46 (9-10) ◽  
pp. 725-734 ◽  
Author(s):  
J. Berlin ◽  
L. Fecker ◽  
C. Rügenhagen ◽  
C. Sator ◽  
D. Strack ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Cell Suspension ◽  
Hairy Root ◽  
Southern Analysis ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Wild Type ◽  
Lupinus Polyphyllus ◽  
Isoflavone Glycoside ◽  
Type Strains

Abstract Transformed cell suspension and hairy root cultures were established by infecting seedlings of Lupinus polyphyllus and L. hartwegii with various wild type strains of Agrobacterium tume-faciens and A. rhizogenes. Transformation of the cultures was confirmed either by their phyto­hormone autotrophy, detection of opines or southern analysis. Glueosides of genistein and 2′-hydroxygenistein, were found to be the main secondary metabolites in normal and trans­formed suspension cultures as well as in hairy root cultures. Although some of the isoflavone glycosides of the cultures were apparently new constituents of Lupinus, they were afterwards also found in young seedlings.

Agrobacterium rhizogenes-Mediated Transformation of Hypericum tomentosum L. and Hypericum tetrapterum Fries.

2009 ◽  
Vol 64 (11-12) ◽  
pp. 864-868 ◽  
Author(s):  
Hedviga Komarovská ◽  
Annalisa Giovannini ◽  
Ján Košuth ◽  
Eva Čellárová
Keyword(s):  
Genetic Transformation ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Wild Type ◽  
Root Segments ◽  
Transgenic Lines ◽  
Type Strains ◽  
Hypericum Species

This is the fi rst evidence on successful Agrobacterium rhizogenes-mediated genetic transformation of two species from the genus Hypericum, H. tomentosum L. and H. tetrapterum Fries. Hairy root cultures were induced from root segments of both Hypericum species by two agropine wild-type strains of A. rhizogenes, ATCC 15834 and A4. The transgenic character of the hairy root cultures was proved by PCR amplifi cation of the rolABCD genes. In some H. tetrapterum transgenic lines aux genes were detected as well.

Biosynthesis of serotonin and β-carboline alkaloids in hairy root cultures of Peganum harmala

1993 ◽  
Vol 33 (3) ◽  
pp. 593-597 ◽  
Author(s):  
Jochen Berlin ◽  
Christiane Rügenhagen ◽  
Norbert Greidziak ◽  
Inna N. Kuzovkina ◽  
Ludger Witte ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Peganum Harmala

MALDI-TOF characterization of hGH1 produced by hairy root cultures ofBrassica oleraceavar.italicagrown in an airlift with mesh bioreactor

2013 ◽  
Vol 30 (1) ◽  
pp. 161-171 ◽  
Author(s):  
Edgar García López ◽  
Emma Gloria Ramos Ramírez ◽  
Octavio Gómez Gúzman ◽  
Graciano Calva Calva ◽  
Armando Ariza-Castolo ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Maldi Tof

Effects of Culture Conditions on Isoflavonoid Levels of Transformed and Non-Transformed Cultures of Lupinus -a Comparison of Suspension and Hairy Root Cultures

1991 ◽  
Vol 46 (9-10) ◽  
pp. 735-742 ◽  
Author(s):  
J. Berlin ◽  
C. Rügenhagen ◽  
M. Rippert ◽  
S. Erdogan
Keyword(s):  
Suspension Culture ◽  
Hairy Root ◽  
Root Culture ◽  
Culture Conditions ◽  
Suspension Cultures ◽  
Stress Factors ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Free Medium ◽  
Lupinus Polyphyllus

Abstract Some highly productive suspension and hairy root cultures were found among several trans­formed cultures of Lupinus polyphyllus and L. hartwegii. A transformed suspension culture Lupo 30150 and a root culture Luha 15834 containing the highest specific isoflavone glucoside content were characterized and compared with normal phytohormone-dependent lines with respect to product stability as well as to their responsiveness to external triggers, e.g. response to changes in the medium. While phytohormone-dependent suspension cultures lost their ini­tial ability to form increased levels of isoflavonoids on phytohormone-free medium, the trans­formed phytohorm one-independent suspension Lupo 30150 remained a highly productive line, despite the fact that its specific levels decreased to 60% of the initial values during several years in liquid medium. Highest stability of product patterns and levels were noted for the transformed root culture. Phytohorm ones had little effect on growth and isoflavonoid levels in suspension cultures, while they reduced both strongly in root cultures. In the presence of 2,4-D the root culture changed into an aggregated low producing suspension culture from which the root state was recovered on phytohorm one-free medium. As long as the root state was main­ tained, isoflavonoid levels could not be distinctly improved by media variation while specific isoflavonoid levels of suspensions were increased by stress factors such as phosphate depletion. When suspensions were transferred to fresh medium phenylalanine ammonia-lyase was greatly induced within 24 h, while the activity remained nearly unchanged in root cultures.

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