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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 38 (1994), S. 189-198 
    ISSN: 1573-5044
    Keywords: Anthranilate synthase ; cell culture ; chorismate mutase ; elicitor induction ; Ruta graveolens ; shikimic acid pathway
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro cultures of Ruta graveolens L. respond with rapid accumulation of acridone epoxides, furoquinolines and furanocoumarins, when challenged with autoclaved homogenate of the yeast Rhodotorula rubra. A transient increase of several enzymes of the respective biosynthetic pathways was measured but we still look for the key regulatory enzymes. We investigated whether the branch point enzymes of the shikimic acid pathway anthranilate synthase (AS) and chorismate mutase (CM) possibly play such a role. The two enzymes compete for chorismate. AS forms anthranilate, the precursor amino acid of acridone and furoquinoline alkaloids. CM channels chorismate into phenylalanine, tyrosine and phenylpropanoid biosynthesis. Elicitation resulted in a transient increase of the activity of both enzymes. Relative induction rates were 2–4 fold for AS and about 1.5 fold for CM. Constitutive CM activity, however, is about 1000 fold higher than AS activity. As in other plants 2 isoforms of CM are expected to be present in R. graveolens. A differential determination of the activity of the isoforms via the tryptophan activation rate proved to be ambiguous. Some evidence for the specific induction of a plastidic form of CM was obtained by inhibition of translation. The time courses of CM induction show CM not to be a key enzyme in elicitor induction of furanocoumarin accumulation. In comparison to other enzyme activities induction of anthranilate synthase activity corresponds closest to inducible acridone epoxide accumulation indicating a key role in its regulation. Induction of AS and CM was inhibited by actinomycin D and chloramphenicol while cycloheximid inhibited AS induction only.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5044
    Keywords: acridone epoxides ; elicitor induction ; N-methyltransferase ; root culture ; Ruta graveolens ; shoot culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Suspension-cultured cells of Ruta graveolens respond with rapid accumulation of acridone epoxides when treated with autoclaved culture homogenate of Rhodotorula rubra. Here the effect of elicitation on S-adenosyl-L-methionine:anthranilic acid N-methyltransferase (NMT), the first specific step in acridone alkaloid biosynthesis, was investigated. In dark-grown and in light-grown cultures, NMT activity remained at low levels (0.1–1 μkat kg-1 protein) over the entire culturing period of 35 days. Elicitation caused a rapid transient increase of NMT activity up to levels of 9–15 μkat kg-1 for light-grown cultures and 16–22 μkat kg-1 for dark-grown ones. Correlation of elicitor dose and induced NMT activity showed saturation rather than an optimum. Culture age and elicitor concentration strongly affected the return of NMT activity to the constitutive level. Addition of inhibitors of transcription or translation suppressed induction of both NMT activity and acridone epoxide accumulation by 50% (actinomycin) and 90–100% (cycloheximide), respectively. Of all the different suspension and organ cultures tested, roots, the site of high constitutive acridone epoxide accumulation, possessed the highest constitutive NMT activities. Different suspension cultures lines, shoot teratomas, hairy root cultures, and roots and shoots of hydroponically and sterile grown plants all showed increased NMT activity upon elicitation. Roots, however, showed the weakest response and, in contrast to all the other systems, induction was not followed by an increase in acridone epoxide content. Tissue-specific acridone epoxides may function as constitutive metabolites or as phytoalexins.
    Type of Medium: Electronic Resource
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