Auxin Conjugation by Tobacco Mesophyll Protoplasts

Caboche, Michel; Aranda, G.; Poll, Annie Marion; Huet, Jean‐Claude; Leguay, Jean-Jacques

doi:10.1104/pp.75.1.54

Cited by 29 publications

(14 citation statements)

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“…Previous work in our laboratory on auxin metabolism described the synthesis of NAA-L-aspartic acid by tobacco protoplasts (10 There is probably no general explanation for the basis ofamino acid toxicity under low density growth conditions. Previous work in our laboratory on valine toxicity has shown that this amino acid was mainly interfering with the biosynthesis of a related amino acid, isoleucine (5).…”

Section: Discussionmentioning

confidence: 99%

“…These experiments were rather tedious to perform at high densities since they required serial countings of colony development under the microscope, and since they had to be performed during short periods of time due to the rapid decrease of NAA concentration in the culture medium promoted by the conjugation processes taking place in the cells (10). These observations allowed us, however, to perform biochemical studies under high density growth conditions, which generally required large amounts of cells.…”

Section: Discussionmentioning

confidence: 99%

“…8). Previous observations have shown that, in contrast to NAA, picloram is poorly conjugated by tobacco mesophyll protoplasts (10). Auxin-Resistant Mutant Clone.…”

Section: Resultsmentioning

confidence: 99%

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Relationship between Auxin and Amino Acid Metabolism of Tobacco Protoplast-Derived Cells

Marion‐Poll

Caboche²

1984

Plant Physiol.

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ABSTRACrSingle amno acids were found to be highly toxic to protoplast-derived cells of tobacco (Nicotiana tabacum cv Xanthi) cultured at low density in a culture medium containing a low naphthaleneacetic acid concentration (0.05 micromolar). The cytotoxicities of alanine, aspartic acid, asparagine, glutanic acid, glutamine, glycine, lysine, proline, and valine were reduced when the naphthaleneacetic acid concentration of the culture medium was increased to I micromolar. This selective modification of amino acid toxicity by naphthaleneacetic acid could not be correlated with modifications of uptake rates or incorporation of these amino acids into protein or amino acid-auxin conjugates. A mutant clone resistant to high naphthaleneacetic acid concentrations and affected in root morphogenesis did not display, at the cellular level, the naphthaleneacetic acidmediated modification of amino acid cytotoxicity.Amino acid toxicity is a well-known phenomenon, studied in whole plants, isolated organs, or cell suspension cultures (3,14,15,20,23,24). Plant cell culture offers the opportunity to study this phenomenon at the cellular level under controlled conditions. Previous studies of amino acid toxicity in tobacco cells gave controversial results. Amino acid toxicity in tobacco cell suspension cultures has been thought to result from the inhibition by amino acids of the first steps of nitrogen metabolism, especially of NR' activity (3,14). However, toxicities of valine and of the mixture lysine plus threonine, in tobacco P-cells, have been shown to be due to specific inhibition of the biosynthesis of related amino acids-respectively, isoleucine and methionine (5, 6)-and not to an inhibition of NR activity (19).One ofthe most important differences between cell suspension and P-cell cultures is the hormone concentration of their culture media. Suspension culture media contain high auxin/cytokinin ratios to prevent clumping, whereas P-cells are generally grown in the presence of low auxin/cytokinin ratios which favor low density growth. In this study, we describe the relationship between auxin concentration of the culture medium and amino acid metabolism of P-cells, cultured at low density. MATERIALS AND METHODSPlant Material, Media, and Culture Conditions. Mesophyll protoplasts were prepared from diploid plants of Nicotiana tabacum (cv Xanthi) and cultured in medium To at a cell density of 7 x I04 protoplasts/ml as previously described (1 1). The same procedure was used to obtain protoplasts from the leaves of the auxin-resistant tobacco clone 36. This mutant clone was obtained after UV-irradiation of haploid protoplasts and selection on a medium containing high NAA concentration. Because mutant plantlets were unable to root, they were grafted on tobacco plants (N. tabacum cv Wisconsin 38) (21; manuscript in preparation).After 4 d ofculture in the dark at 26C, the P-cells were washed in nitrogen-and auxin-free medium C (22). Cells were then diluted to low density (102 divided cells/ml) in glutamine-free medium C containing m...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Relationship between Auxin and Amino Acid Metabolism of Tobacco Protoplast-Derived Cells

Marion‐Poll

Caboche²

1984

Plant Physiol.

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“…one-half of the esters are the conjugates of IAA and cellulosic glucans (Piskornik and Bandurski 1972). The ester conjugates seem to predominate among monocotyledon plants (maize, rice), though their synthesis has also been found in Arabidopsis (Tam et al 2000), immature bean seeds (Bialek and Cohen 1989;Jakubowska and Kowalczyk 2004), Scots pine seeds (Ljung et al 2001), tomato pericarp (Catalá et al 1992), henbane cells (Oetiker and Aeschbacher 1997) or tobacco leaf protoplasts (Caboche et al 1984;Delbarre et al 1994).…”

Section: Introductionmentioning

confidence: 99%

Novel 1-O-indole-3-acetyl-β-d-glucose-dependent acyltransferase transferring indoleacetyl moiety to some mono- di- and oligosaccharides

Starzyńska

Kowalczyk

2011

Acta Physiol Plant

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“…studies (7). A genetic approach to the study ofauxin action could help in understanding this process ofauxin-mediated cell killing.…”

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confidence: 99%

Comparison of the Growth Promoting Activities and Toxicities of Various Auxin Analogs on Cells Derived from Wild Type and a Nonrooting Mutant of Tobacco

Caboche¹,

Müller²,

Chanut³

et al. 1987

Plant Physiol.

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The aim of the present work was to further study the characteristics of stimulation of growth of wild type and mutant cells by various auxins. The ability of the mutant to form auxinconjugates was investigated as a possible basis for increased auxin tolerance. An auxin analog specifically triggering cell death was identified by differential screening on wild-type and mutant cells. MATERIALS AND METHODSAn auxin requirement for the stimulation of cell proliferation has been demonstrated for many types of tissues grown in vitro. Recent work on the transformation of tissues by Agrobacterium tumefaciens have shown that the oncogenous transferred bacterial DNA encodes for two genes involved in a pathway of auxin biosynthesis, thus abolishing the requirement of exogenously supplied auxin for the growth of transformed cells (13,22). The mechanism by which auxin triggers cell proliferation and differentiation is still far from being understood. Specific auxin receptors have been postulated but no proof for their function in mediating auxin actions has been given (1 1).Cells derived from mesophyll protoplasts of tobacco and grown at low densities were found to be killed by relatively low auxin concentrations (5 FAM NAA2 or IAA) (6). A correlation between such toxicity and auxin conjugation was also observed in these 'Supported in part by a grant from the Ministere de l'Industrie et de la Recherche, "GEn&tique et physiologie des veg6taux sup6rieurs." 2Abbreviations: NAA, naphthaleneacetic acid; DAST, diethylaminosulfur trifluoride; P-cell, protoplast-derived cell; PE, plating efficiency; picloram, 3,5,6-trichloropicolinic acid; RPE, relative plating efficiency.

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Auxin Conjugation by Tobacco Mesophyll Protoplasts

Cited by 29 publications

References 14 publications

Relationship between Auxin and Amino Acid Metabolism of Tobacco Protoplast-Derived Cells

Relationship between Auxin and Amino Acid Metabolism of Tobacco Protoplast-Derived Cells

Novel 1-O-indole-3-acetyl-β-d-glucose-dependent acyltransferase transferring indoleacetyl moiety to some mono- di- and oligosaccharides

Comparison of the Growth Promoting Activities and Toxicities of Various Auxin Analogs on Cells Derived from Wild Type and a Nonrooting Mutant of Tobacco

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