Quantification of indole-3-acetic acid in untransformed and Agrobacterium rhizogenes-transformed pea roots using gas chromatography mass spectrometry

Schaerer, Santiago; Pilet, Paul‐Emile

doi:10.1007/bf00201343

Cited by 9 publications

(3 citation statements)

References 29 publications

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“…Our results were consistent with data showing that hairy roots from bittersweet nightshade (Gartland et al, 1991) and pea (Schaerer and Pilet, 1993) and shoots from TL-DNAtransformed rape seed (Julliard et al, 1992) display normal levels of endogenous IAA. More specifically, our results agreed with the recent evidence that the expression of the 35s rolB gene in tobacco plants has no significant effect on either the endogenous levels of IAA (Nilsson et al, 1993;Schmiilling et al, 1993) or the rate of IAA metabolism or the in vivo hydrolysis of exogenously supplied IAA Glc ester (Nilsson et al, 1993).…”

Section: Dlscusslonsupporting

confidence: 92%

The rolB Gene of Agrobacterium rhizogenes Does Not Increase the Auxin Sensitivity of Tobacco Protoplasts by Modifying the Intracellular Auxin Concentration

et al. 1994

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Phenotypical alterations observed in rolB-transformed plants have been proposed to result from a rise in intracellular free auxin due to a RolB-catalyzed hydrolysis of auxin conjugates (J.J. Estruch, J. Schell, A. Spena [1991] EMBO J 10: 3125-3128). We have investigated this hypothesis in detail using tobacco (Nicofiana tabacum) mesophyll protoplasts isolated from plants transformed with the rolB gene under the control of its own promoter (BBCUS 6 clone) or the cauliflower mosaic virus 35s promoter (CaMVBT 3 clone). Protoplasts expressing rolB showed an increased sensitivity to the auxin-induced hyperpolarization of the plasma membrane when triggered with exogenous auxin. Because this phenotypical trait was homogeneously displayed over the entire population, protoplasts were judged to be a more reliable test system than the tissue fragments used in previous studies to monitor rolB gene effects on cellular auxin levels. Accumulation of free 1-[3H]-naphthaleneacetic acid (NAA) was equivalent in CaMVBT 3, BBCUS 6, and wild-type protoplasts. Naphthyl-B-glucose ester, the major NAA metabolite in protoplasts, reached similar levels in CaMVBT 3 and normal protoplasts and was hydrolyzed at the same rate in BBCUS 6 and normal protoplasts. Furthermore, NAA accumulation and metabolism in BBCUS 6 protoplasts were independent of the rolB gene expression level. Essentially similar results were obtained with indoleacetic acid. Thus, i t was concluded that the rolB-dependent behavior of transgenic tobacco protoplasts is not a consequence of modifying the intracellular auxin concentration but likely results from changes in the auxin perception pathway.

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Section: Dlscusslonsupporting

confidence: 92%

The rolB Gene of Agrobacterium rhizogenes Does Not Increase the Auxin Sensitivity of Tobacco Protoplasts by Modifying the Intracellular Auxin Concentration

et al. 1994

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“…The enhanced content of endogenous IAA in transgenic plants may be due to the expression of aux and rol genes of A. rhizogenes. Similar results were reported in Pisum sativum, where an agropine-type hairy root induced by A. rhizogenes strain 15834 contains significantly higher concentration of IAA than other transformed root lines and non-transformed roots (Schaerer and Pilet, 1992). They further reported that, the IAA content was very small in untransformed and transformed root lines, compared with the levels in untransformed intact primary roots.…”

Section: Endogenous Iaa Concentrationsupporting

confidence: 73%

Production of transgenic Aralia elata regenerated from Agrobacterium rhizogenes-mediated transformed roots

Kang

Anbazhagan

You

et al. 2006

Plant Cell Tiss Organ Cult

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Transgenic hairy roots were induced from petiole and root segments of in vitro plant Aralia elata, a medicinal woody shrub, after co-cultivation with A. rhizogenes ATCC 15834. The percentage of putative hairy root induction from root segments was higher (26.7%) than petiole explants (10.0%). Hairy roots showed active production of lateral roots with vigorous elongation. Transgenic plants were regenerated from hairy roots via somatic embryogenesis. These plants had wrinkled leaves, short petioles and numerous lateral hairy roots. The RT-PCR analysis showed the expression of rol A, B, C, D, aux 1 and 2 genes differed between the transgenic lines. Endogenous IAA level was higher in transgenic than non-transgenic plants. Conclusively, transgenic hairy roots were developed for first time in A. elata and the transgenic hairy root lines showed distinct morphological growth pattern and gene expression.

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“…The levels of free IAA in N and T roots of H. muticus are comparatively low. They are, however, in line with data published by Schaerer and Pilet (1993) where long term root cultures of Pisum sativum, rather than newly initiated ones, were studied. In spite of their growth rate and prolific branching, all hairy root lines except one had lower IAA levels than intact primary roots.…”

Section: Treatmentsupporting

confidence: 71%

Hormonal Effects on Growth and Morphology of Normal and Hairy Roots of Hyoscyamus muticus

Biondi

Lenzi

Baraldi

et al. 1997

J Plant Growth Regul

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Treatment of normal and Agrobacterium rhizogenes-transformed root cultures of Hyoscyamus muticus with three different auxins, indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and naphthaleneacetic acid (NAA), revealed that the response varied considerably among auxins, between transformed and normal roots, and depending on the parameter. In normal roots all three auxins provoked abundant branching, with IBA and NAA being the most effective at 2.5 and 0.5 M, respectively, whereas IAA was most effective at low concentrations (0.05 and 0.1 M). In transformed roots exogenously supplied auxins were generally inhibitory or, at best, without effect on growth and branching. Only 0.01 M IAA significantly enhanced lateral root number, whereas at the higher concentrations IBA, although inhibitory, was the least effective auxin. In both root types IBA had little effect on primary root growth, but normal roots were more sensitive to IAA and NAA. These results suggest a different sensitivity to auxins of normal and transformed roots since there was no significant difference in endogenous free and conjugated IAA content nor in IAA uptake capacity. Ethylene production and biosynthesis were approximately threefold higher in hairy roots, but production could be stimulated up to tenfold that of control levels in normal roots by supplying NAA or 1-aminocyclopropane-1-carboxylic acid (ACC). Treatment with 2.5 M NAA, but not IAA or IBA, also enhanced ethylene biosynthesis in normal roots but not in transformed ones. ACC and malonyl-1-aminocyclopropane-1-carboxylic acid accumulated to detectable levels only after treatment with an auxin (NAA).

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Quantification of indole-3-acetic acid in untransformed and Agrobacterium rhizogenes-transformed pea roots using gas chromatography mass spectrometry

Cited by 9 publications

References 29 publications

The rolB Gene of Agrobacterium rhizogenes Does Not Increase the Auxin Sensitivity of Tobacco Protoplasts by Modifying the Intracellular Auxin Concentration

The rolB Gene of Agrobacterium rhizogenes Does Not Increase the Auxin Sensitivity of Tobacco Protoplasts by Modifying the Intracellular Auxin Concentration

Production of transgenic Aralia elata regenerated from Agrobacterium rhizogenes-mediated transformed roots

Hormonal Effects on Growth and Morphology of Normal and Hairy Roots of Hyoscyamus muticus

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