Cytochrome P450 CYP79B2 from Arabidopsis catalyzes the conversion of tryptophan to indole-3-acetaldoxime, a precursor of indole glucosinolates and indole-3-acetic acid.
(73/2030)
Glucosinolates are natural plant products known as flavor compounds, cancer-preventing agents, and biopesticides. We report cloning and characterization of the cytochrome P450 CYP79B2 from Arabidopsis. Heterologous expression of CYP79B2 in Escherichia coli shows that CYP79B2 catalyzes the conversion of tryptophan to indole-3-acetaldoxime. Recombinant CYP79B2 has a K(m) of 21 microm and a V(max) of 7.78 nmol/h/ml culture. Inhibitor studies show that CYP79B2 is different from a previously described enzyme activity that converts tryptophan to indole-3-acetaldoxime (Ludwig-Muller, J. , and Hilgenberg, W. (1990) Phytochemistry, 29, 1397-1400). CYP79B2 is wound-inducible and expressed in leaves, stem, flowers, and roots, with the highest expression in roots. Arabidopsis overexpressing CYP79B2 has increased levels of indole glucosinolates, which strongly indicates that CYP79B2 is involved in indole glucosinolate biosynthesis. Our data show that oxime production by CYP79s is not restricted to those amino acids that are precursors for cyanogenic glucosides. Our data are consistent with the hypothesis that indole glucosinolates have evolved from cyanogenesis. Indole-3-acetaldoxime is a precursor of the plant hormone indole-3-acetic acid, which suggests that CYP79B2 might function in biosynthesis of indole-3-acetic acid. Identification of CYP79B2 provides an important tool for modification of the indole glucosinolate content to improve nutritional value and pest resistance. (+info)
A role for the rice homeobox gene Oshox1 in provascular cell fate commitment.
(74/2030)
The vascular tissues of plants form a network of interconnected cell files throughout the plant body. The transition from a genetically totipotent meristematic precursor to different stages of a committed procambial cell, and its subsequent differentiation into a mature vascular element, involves developmental events whose molecular nature is still mostly unknown. The rice protein Oshox1 is a member of the homeodomain leucine zipper family of transcription factors. Here we show that the strikingly precise onset of Oshox1 gene expression marks critical, early stages of provascular ontogenesis in which the developmental fate of procambial cells is specified but not yet stably determined. This suggests that the Oshox1 gene may be involved in the establishment of the conditions required to restrict the developmental potential of procambial cells. In support of this hypothesis, ectopic expression of Oshox1 in provascular cells that normally do not yet express this gene results in anticipation of procambial cell fate commitment, eventually culminating in premature vascular differentiation. Oshox1 represents the first example of a transcription factor whose function can be linked to specification events mediating provascular cell fate commitment. (+info)
Early physiological and cytological events induced by wounding in potato tuber.
(75/2030)
The response of potato tuber (Solanum tuberosum L. cv. Kennebec) to mechanical wounding was investigated at different times. Changes in the levels of indole-3-acetic acid (IAA), polyunsaturated fatty acids (PUFAs) and lipid hydroperoxides (LOOHs) were monitored up to 120 min after wounding and related to the cytological events occurring up to 24 h. Twenty minutes after injury, an increase in IAA and LOOH levels and a decrease in the levels of PUFAs was observed. Wounding induced mitoses in differentiated (parenchyma) cells starting at 120 min, and promoted an increase of mitotic activity in the meristematic cells (procambium and bud dome), after 360 min. The inhibition of the increase in LOOHs and IAA by lipoxygenase (LOX) inhibitors, as well as the ability of in vitro peroxidated linoleic acid to enhance IAA production, suggest a close relationship among lipoperoxidation, IAA and mitotic activity in the response of potato tuber cells to injury, resulting in a specific growth response, i.e. bud growth and periderm formation. (+info)
Limited correlation between expansin gene expression and elongation growth rate.
(76/2030)
The aim of this work was to study the role of the cell wall protein expansin in elongation growth. Expansins increase cell wall extensibility in vitro and are thought to be involved in cell elongation. Here, we studied the regulation of two tomato (Lycopersicon esculentum cv Moneymaker) expansin genes, LeExp2 and LeExp18, in rapidly expanding tissues. LeExp2 was strongly expressed in the elongation zone of hypocotyls and in the faster growing stem part during gravitropic stimulation. LeExp18 expression did not correlate with elongation growth. Exogenous application of hormones showed a substantial auxin-stimulation of LeExp2 mRNA in etiolated hypocotyls and a weaker auxin-stimulation of LeExp18 mRNA in stem tissue. Analysis of transcript accumulation revealed higher levels of LeExp2 and LeExp18 in light-treated, slow-growing tissue than in dark-treated, rapidly elongating tissue. Expansin protein levels and cell wall extension activities were similar in light- and dark-grown hypocotyl extracts. The results show a strong correlation between expansin gene expression and growth rate, but this correlation is not absolute. We conclude that elongation growth is likely to be controlled by expansin acting in concert with other factors that may limit growth under some physiological conditions. (+info)
Regulation of Arabidopsis root development by nitrate availability.
(77/2030)
When the root systems of many plant species are exposed to a localized source of nitrate (NO3- they respond by proliferating their lateral roots to colonize the nutrient-rich zone. This study reviews recent work with Arabidopsis thaliana in which molecular genetic approaches are being used to try to understand the physiological and genetic basis for this response. These studies have led to the conclusion that there are two distinct pathways by which NO3- modulates root branching in Arabidopsis. On the one hand, meristematic activity in lateral root tips is stimulated by direct contact with an enriched source of NO3- (the localized stimulatory effect). On the other, a critical stage in the development of the lateral root (just after its emergence from the primary root) is highly susceptible to inhibition by a systemic signal that is related to the amount of NO3- absorbed by the plant (the systemic inhibitory effect). Evidence has been obtained that the localized stimulatory effect is a direct effect of the NO3- ion itself rather than a nutritional effect. A NO3(-)-inducible MADS-box gene (ANR1) has been identified which encodes a component of the signal transduction pathway linking the external NO3- supply to the increased rate of lateral root elongation. Experiments using auxin-resistant mutants have provided evidence for an overlap between the auxin and NO3- response pathways in the control of lateral root elongation. The systemic inhibitory effect, which does not affect lateral root initiation but delays the activation of the lateral root meristem, appears to be positively correlated with the N status of the plant and is postulated to involve a phloem-mediated signal from the shoot. (+info)
Auxin-dependent cell wall depositions in the epidermal periplasmic space of graviresponding nodes of Tradescantia fluminensis.
(78/2030)
Differential growth of the nodal regions of graviresponding Tradescantia fluminensis (Wandering Jew) was analysed with special respect to the extension-restricting epidermal cells of the opposite growing and growth-inhibited organ flanks. Gravicurvature of horizontally gravistimulated isolated nodes depends on auxin (indolyl-3-acetic acid, IAA) and shows a node-specific profile in which the third node below the tip showed the greatest response. Exogenously supplied gibberellic acid induced no gravitropic growth. Vertically oriented isolated nodes supplied with exogenous IAA showed, on an electron microscopical level, conspicuous membrane invaginations with adjacent wall depositions restricted to the outer tangential epidermal cell walls. Their number was more than doubled by exogenously supplied Ca2+, which inhibited IAA-induced growth. No such changes could be detected in water-incubated segments or inner tissues of IAA-supplied segments. Gravistimulated differential growth of nodes of intact shoots and of nodal segments was characterized by changes similar to the ones induced by exogenous IAA, with greatly increased numbers of wall depositions within the epidermal cells of the growth-inhibited upper organ flank. Similar to the gravistimulated wall depositions, an asymmetric distribution pattern of Ca2+ was detected in the epidermal cell walls employing x-ray energy spectrum analysis (EDX). The results indicate that growth of nodes of Tradescantia fluminensis is regulated via IAA-induced secretion and subsequent infiltration of wall components enabling wall extension. The data support the hypothesis that temporary differential growth during gravicurvature of Tradescantia fluminensis is mediated by the antagonistic effect of Ca(2+)-ions on the infiltration of IAA-induced wall-loosening components into the outer, extension-restricting epidermal walls thereby inhibiting growth. (+info)
Low temperature sensing in tulip (Tulipa gesneriana L.) is mediated through an increased response to auxin.
(79/2030)
Tulip (Tulipa gesneriana L.) is a bulbous plant species that requires a period of low temperature for proper growth and flowering. The mechanism of sensing the low temperature period is unknown. The study presented in this paper shows that the essential developmental change in tulip bulbs during cold treatment is an increase in sensitivity to the phytohormone auxin. This is demonstrated using a model system consisting of isolated internodes grown on tissue culture medium containing different combinations of the phytohormones auxin and gibberellin. Using mathematical modelling, equations taken from the field of enzyme kinetics were fitted through the data. By doing so it became apparent that longer periods of low temperature resulted in an increased maximum response at a lower auxin concentration. Besides the cold treatment, gibberellin also enhances the response to auxin in the internodes in this in vitro system. A working model describing the relationship between the cold requirement, gibberellin action and auxin sensitivity is put forward. Possible analogies with other cold-requiring processes such as vernalization and stratification, and the interaction of auxin and gibberellin in the stalk elongation process in other plant species are discussed. (+info)
Isolation and characterization of low-indole-3-acetic acid-producing mutants from Bradyrhizobium elkanii.
(80/2030)
We isolated 11 low-indole-3-acetic acid (IAA)-producing mutants of Bradyrhizobium elkanii by Tn5 mutagenesis. The amount of IAA produced by each mutant was 2.2-13.6% of that of the wild-type. It was found by resting cell reactions that the biosynthetic step to convert indole-3-pyruvic acid to indole-3-acetaldehyde was blocked in all the mutants. (+info)