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Molecular approaches to understanding auxin action.

The plant hormone auxin is thought to be involved in diverse growth and developmental processes. The mechanisms by which the hormone exerts its effects are unknown. Recent evidence suggests that auxin can rapidly and specifically alter the expression of a select set of mRNAs. Auxin-responsive mRNAs from several plant systems have been isolated and characterized and, in a few cases, the corresponding gene sequence and organization have been elucidated. The rapid accumulation of auxin-responsive mRNAs appears to be regulated at both the transcriptional and post-transcriptional levels. Research efforts are currently focused on elucidating the cis-acting elements and trans-acting factors that mediate the regulation of genes by auxin. Additionally, the function of the auxin-responsive gene products and their relationship, if any, to auxin-regulated growth processes remain to be determined.

Gene Expression Regulation↗

Cell-to-cell communication via plasmodesmata during Arabidopsis embryogenesis.

In Arabidopsis embryogenesis, positional information establishes the overall body plan and lineage-dependent cell fate specifies local patterning. Position-dependent gene expression and responses to the plant hormone auxin are also crucial. Recently, another mechanism that delivers positional information has been uncovered. This pathway utilizes cell-to-cell communication via plasmodesmata. Plasmodesmata span the walls between neighboring plant cells. Groups of cells that allow intercellular transport of biotic and abiotic tracers form symplastic domains of shared communication. Initially, cells of the embryo form one symplast. As development proceeds, symplastic sub-domains that correspond to the major morphological regions of the plant (i.e. shoot apex, cotyledons, hypocotyl, and root) are formed. These sub-domains further resolve into tissue-specific domains of communication (such as protodermal and vascular regions). Cell-to-cell communication via plasmodesmata between embryonic and maternal tissues ceases as development proceeds.

Arabidopsis↗

Control of auxin-regulated root development by the Arabidopsis thaliana SHY2/IAA3 gene.

The plant hormone auxin controls many aspects of development and acts in part by inducing expression of various genes. Arabidopsis thaliana semidominant shy2 (short hypocotyl) mutations cause leaf formation in dark-grown plants, suggesting that SHY2 has an important role in regulating development. Here we show that the SHY2 gene encodes IAA3, a previously known member of the Aux/IAA family of auxin-induced genes. Dominant shy2 mutations cause amino acid changes in domain II, conserved among all members of this family. We isolated loss-of-function shy2 alleles including a putative null mutation. Gain-of-function and loss-of-function shy2 mutations affect auxin-dependent root growth, lateral root formation, and timing of gravitropism, indicating that SHY2/IAA3 regulates multiple auxin responses in roots. The phenotypes suggest that SHY2/IAA3 may activate some auxin responses and repress others. Models invoking tissue-specificity, feedback regulation, or control of auxin transport may explain these results.

Amino Acid Sequence↗

Regulation of plant ferritin synthesis: how and why.

Plant ferritins are key iron-storage proteins that share important structural and functional similarities with animal ferritins. However, specific features characterize plant ferritins, among which are plastid cellular localization and transcriptional regulation by iron. Ferritin synthesis is developmentally and environmentally controlled, in part through the differential expression of the various members of a small gene family. Furthermore, a strict requirement for plant ferritin synthesis regulation is attested to by alterations of the photosynthetic apparatus and of iron homeostasis in transgenic tobaccos overexpressing these proteins. Plant ferritin gene regulation appears to consist of a complex interplay of transcriptional and posttranscriptional mechanisms, involving cellular relays such as plant hormones, oxidative steps and Ser/Thr phosphatase.

Animals↗

Dispatch. Gibberellin signaling: GRASs growing roots dispatch.

Gibberellins are important endogenous plant growth regulators. A new study has revealed an unexpected role for gibberellin-regulated transcription factors in promoting root growth; this depends on signaling from another plant hormone, auxin, suggesting that auxin promotes root growth by modulating the cellular response to gibberellins.

Gibberellins↗

Effects of erythromycin, tetracycline and ibuprofen on the growth of Synechocystis sp. and Lemna minor.

Pharmaceutically active substances have recently been recognised as an emerging environmental problem. Human and veterinarian therapeutic agents can contaminate aquatic ecosystems via sewage discharges (human and animal excretion), improper disposal or industrial waste. Very little is known on the effects of pharmaceutical pollutants on aquatic photosynthetic organisms. In this study the effects of erythromycin, tetracycline and ibuprofen on the growth of the cyanobacterium Synechocystis sp. PCC6803 and the duckweed Lemna minor FBR006 were studied at concentrations of 1-1000 microg l(-1). At dosage of 1 mg l(-1), erythromycin affected the growth of both Synechocystis and Lemna with a maximum inhibition of 70 and 20%, respectively. Tetracycline had inhibitory effects (20-22% reduction in growth) on Synechocystis at intermediate dosages. The same aminoglycoside antibiotic promoted growth in Lemna by 26% at 10 microg l(-1), while frond development was reduced at 1 mg l(-1) (tetracycline). The anti-inflammatory ibuprofen strongly stimulated the growth of Synechocystis at all concentrations tested (72% increase at 10 microg l(-1)) although inhibited Lemna in a linear dose-dependent manner with a 25% reduction over control levels at a dosage of 1 mg l(-1). The 7 days effective concentration (EC(50)) calculated for Lemna were 5.6, 1 and 4 g l(-1), respectively, for erythromycin, tetracycline and ibuprofen. Moreover, exposure to the three pharmaceuticals resulted in the production of the stress hormone, abscisic acid (ABA), in Lemna. Erythromycin and tetracycline were more effective in promoting ABA synthesis compared to ibuprofen. The effects shown by the three therapeutic drugs on Synechocystis and Lemna growth may have potential implications in the assessments of residual environmental risks associated with the presence of pharmaceuticals in freshwater ecosystems. Promotion of ABA synthesis in Lemna by the two antibiotics and by copper suggests that the plant hormone could be a suitable (additional) indicator for future evaluation of phytotoxicity that results in plant senescence.

Abscisic Acid↗

Hormone interactions and regulation of Unifoliata, PsPK2, PsPIN1 and LE gene expression in pea (Pisum sativum) shoot tips.

The Unifoliata (Uni) gene plays a major role in development of the compound leaf in pea, but its regulation is unknown. In this study, we examined the effects of plant hormones on the expression of Uni, PsPK2 (the gene for a pea homolog of Arabidopsis PID, a regulator of PIN1 targeting), PsPIN1 (the major gene for a putative auxin efflux carrier) and LE (a gibberellin biosynthesis gene, GA3ox), and also examined mutual hormonal regulation of these genes, in pea shoot tips, including a number of mutants. The Uni promoter possessed putative auxin and gibberellin response elements. The PsPIN1 mRNA levels were increased in afila, which replaces leaflets with branched tendrils; and reduced in tendrilless, which replaces tendrils with leaflets, compared with the wild type (WT). In contrast, mRNA levels of LE were increased in uni and tendrilless and decreased in afila compared with the WT. Uni, PsPK2 and PsPIN1 are positively regulated by gibberellin and auxin, and were induced to higher levels by simultaneous application of auxin and gibberellin. Auxin induction of Uni, PsPK2 and PsPIN1 did not require de novo protein synthesis. LE was positively regulated by auxin and cytokinin. In conclusion, these results support the hypothesis that auxin and gibberellin positively regulate Uni, which controls pea compound leaf development. Also, Uni, PsPIN1, PsPK2 and LE are expressed differentially in the leaf mutants, suggesting that mutual regulation by auxin and gibberellin promotes compound leaf development.

Cytokinins↗

The in vitro effects of opines and other compounds on DNAs originating from bacteria, and from healthy and tumorous plant tissues.

Purified total DNAs were isolated from oncogenic or nononcogenic Agrobacterium tumefaciens cells as well as from normal and crown gall tissues. Opines (octopine, nopaline, lysopine), plant hormone (auxin IAA) and some carcinogenic compounds were used in order to correlate their effects on in vitro strand separation and synthesis of DNAs with in vivo tumorous cell multiplication. Octopine (or nopaline) induced chain opening of DNAs originating from octopine (or nopaline)-metabolizing bacteria and from same bacteria strain-induced tumorous cells. This phenomenon was measured by the increase in DNA hyperchromicity which is concentration dependent. The tested compounds stimulated the in vitro synthesis of the same DNAs. Under the same conditions, in vitro strand separation and synthesis of healthy plant DNA was not (or only slightly) enhanced, except in the case of particular hormone-connected healthy cell DNA. IAA and carcinogens stimulated in vitro synthesis and induced in vitro strand separation (dose-dependent effect) of DNAs isolated from crown gall cells and inducing bacteria. Compared to healthy cell DNAs, these DNAs were thus susceptible to structurally very diversified molecules and in this way behave as do mammalian tissue DNAs. The opine and IAA actions observed here were specific for plant tissue DNA; cancerous human or animal tissue DNAs were insensitive. By their presence in the crown gall cells, opines possibly maintain destabilized areas (required for rapid growth and division) on tumor cell DNA. The cooperative actions of IAA and opines as well as small RNA and RNA fragments on gene activation, might explain the autonomy of plant tumor cells.

9,10-Dimethyl-1,2-benzanthracene↗

Reactive oxygen species as signals that modulate plant stress responses and programmed cell death.

Reactive oxygen species (ROS) are known as toxic metabolic products in plants and other aerobic organisms. An elaborate and highly redundant plant ROS network, composed of antioxidant enzymes, antioxidants and ROS-producing enzymes, is responsible for maintaining ROS levels under tight control. This allows ROS to serve as signaling molecules that coordinate an astonishing range of diverse plant processes. The specificity of the biological response to ROS depends on the chemical identity of ROS, intensity of the signal, sites of production, plant developmental stage, previous stresses encountered and interactions with other signaling molecules such as nitric oxide, lipid messengers and plant hormones. Although many components of the ROS signaling network have recently been identified, the challenge remains to understand how ROS-derived signals are integrated to eventually regulate such biological processes as plant growth, development, stress adaptation and programmed cell death.

Apoptosis↗

Abscisic acid is involved in the iron-induced synthesis of maize ferritin.

The ubiquitous iron storage protein ferritin has a highly conserved structure in plants and animals, but a distinct cytological location and a different level of control in response to iron excess. Plant ferritins are plastid-localized and transcriptionally regulated in response to iron, while animal ferritins are found in the cytoplasm and have their expression mainly controlled at the translational level. In order to understand the basis of these differences, we developed hydroponic cultures of maize plantlets which allowed an increase in the intracellular iron concentration, leading to a transient accumulation of ferritin mRNA and protein (Lobréaux,S., Massenet,O. and Briat,J.F., 1992, Plant Mol. Biol., 19, 563-575). Here, it is shown that iron induces ferritin and RAB (Responsive to Abscisic Acid) mRNA accumulation relatively with abscisic acid (ABA) accumulation. Ferritin mRNA also accumulates in response to exogenous ABA. Synergistic experiments demonstrate that the ABA and iron responses are linked, although full expression of the ferritin genes cannot be entirely explained by an increase in ABA concentration. Inducibility of ferritin mRNA accumulation by iron is dramatically decreased in the maize ABA-deficient mutant vp2 and can be rescued by addition of exogenous ABA, confirming the involvement of ABA in the iron response in plants. Therefore, it is concluded that a major part of the iron-induced biosynthesis of ferritin is achieved through a pathway involving an increase in the level of the plant hormone ABA. The general conclusion of this work is that the synthesis of the same protein in response to the same environmental signal can be controlled by separate and distinct mechanisms in plants and animals.

Abscisic Acid↗

Characterization of a novel carotenoid cleavage dioxygenase from plants.

The plant hormone abscisic acid is derived from the oxidative cleavage of a carotenoid precursor. Enzymes that catalyze this carotenoid cleavage reaction, nine-cis epoxy-carotenoid dioxygenases, have been identified in several plant species. Similar proteins, whose functions are not yet known, are present in diverse organisms. A putative cleavage enzyme from Arabidopsis thaliana contains several highly conserved motifs found in other carotenoid cleavage enzymes. However, the overall homology with known abscisic acid biosynthetic enzymes is low. To determine the biochemical function of this protein, it was expressed in Escherichia coli and used for in vitro assays. The recombinant protein was able to cleave a variety of carotenoids at the 9-10 and 9'-10' positions. In most instances, the enzyme cleaves the substrate symmetrically to produce a C(14) dialdehyde and two C(13) products, which vary depending on the carotenoid substrate. Based upon sequence similarity, orthologs of this gene are present throughout the plant kingdom. A similar protein in beans catalyzes the same reaction in vitro. The characterization of these activities offers the potential to synthesize a variety of interesting, natural products and is the first step in determining the function of this gene family in plants.

Abscisic Acid↗

Compact shoot and leafy head 1, a mutation affects leaf initiation and developmental transition in rice (Oryza sativa L).

The shoot apical meristem (SAM) produces lateral organs in a regular spacing (phyllotaxy) and at a regular interval (phyllochron) during the vegetative phase. In a Dissociation (Ds) insertion rice population, we identified a mutant, compact shoot and leafy head 1 (csl1), which produced massive number of leaves (~70) during the vegetative phase. In csl1, the transition from the vegetative to the reproductive phase was delayed by about 2 months under long-day conditions. With a reduced leaf size and severe dwarfism, csl1 failed to produce a normal panicle after the transition to reproductive growth. Instead, it produced a leafy panicle, in which all primary rachis-branches were converted to vegetative shoots. Phenotypically csl1 resembled pla mutants in short plastochron but was more severe in the conversion of the reproductive organs to vegetative organs. In addition, neither the expression nor the coding region of PLA1 or PLA2 was affected in csl1. csl1 is most likely a dominant mutation because no mutant segregant was observed in progeny of 67 siblings of the csl1 mutant. CSL1 may represent a novel gene, which functions downstream of PLA1 and/or PLA2, or alternatively functions in a separate pathway, involved in the regulation of leaf initiation and developmental transition via plant hormones or other mobile signals.

Gene Expression Regulation, Developmental↗

Discoveries and Dilemmas Concerning Cytokinin Metabolism.

Natural cytokinin compounds that function as plant hormones are adenine molecules connected to an N6 side chain. Metabolic pathways produce various structural modifications to the adenine moiety and/or the side chain that affect cytokinin function, activity, stability, and transport in plants. This review focuses on recent research that is elucidating the genes and biochemical pathways involved in cytokinin conjugation, deconjugation, and interconversion. One area of new discovery concerns the process of O-glucoside conjugate formation. Genes have been identified for O-glucosyltransferase enzymes in Phaseolus (ZOG1 and ZOX1) and maize (cisZOG1) and have been used to study enzyme activity, structure, and function in plant development. New information about the genetic basis of the adenine salvage pathway and its relationship to cytokinin interconversion and regulation has also become available. However, various dilemmas about cytokinin metabolism remain and need to be resolved using a combination of research approaches.

Journal Article↗

How canalization can make loops: a new model of reticulated leaf vascular pattern formation.

Formation of the vascular system in plant leaves can be explained by the canalization hypothesis which states that veins are formed in an initially homogeneous field by a self-organizing process between the plant hormone auxin and auxin carrier proteins. Previous models of canalization can generate vein patterns with branching but fail to generate vein patterns with closed loops. However, closed vein loops are commonly observed in plant leaves and are important in making them robust to herbivore attacks and physical damage. Here we propose a new model which generates a vein system with closed loops. We postulate that the "flux bifurcator" level is enhanced in cells with a high auxin flux and that it causes reallocation of auxin carriers toward neighbouring cells also having a high bifurcator level. This causes the auxin flux to bifurcate, allowing vein tips to attach to other veins creating vein loops. We explore several alternative functional forms for the flux bifurcator affecting the reallocation of efflux carriers and examine parameter dependence of the resulting vein pattern.

Indoleacetic Acids↗

Environmental stress response in plants: the role of mitogen-activated protein kinases.

Mitogen-activated protein kinase (MAPK) cascades have essential roles in diverse intracellular signaling processes in plants, animals and yeasts. In plants, transcription of genes encoding protein kinases involved in MAPK cascades is upregulated by environmental stresses and plant hormones; in addition, MAPK-like kinase activities are transiently activated in response to environmental stresses. Consequently, MAPK cascades are now thought to have important roles in stress signal transduction pathways in higher plants.

Phosphorylation↗

Characterization of two cDNAs (ERD10 and ERD14) corresponding to genes that respond rapidly to dehydration stress in Arabidopsis thaliana.

Two cDNA clones, designated ERD10 and EDR14, isolated from a cDNA library from one-hour-dehydrated plants of Arabidopsis thaliana L. were sequenced and characterized. The predicted ER10 and ERD14 polypeptides have a compositional bias towards Glu (19.62% and 21.08%, respectively) and Lys (16.15% and 18.38%, respectively) and both lack Trp and Cys residues. The amino acid sequences deduced from of both cDNAs are very similar to those of Group II LEA (late embryogenesis abundant) proteins, a class of proteins that accumulate late in embryogenesis and whose gene expression is induced by the plant hormone abscisic acid (ABA). Genomic Southern blot analysis suggested the existence of an ERD10-related gene. The expression of the genes that correspond to ERD10 and ERD14 was strongly induced in rosette plants of Arabidopsis within 1 h of dehydration and elevated levels of transcripts were detected during 24 h of dehydration. Northern blot analysis of the cold-induced expression of both the erd10 and erd14 genes revealed a two-step induction process. Early induction occurred within 1 h and secondary induction occurred 5 h after the beginning of cold stress. Application of ABA induced the expression of both genes, but application of 2,4-D, BA, ABA and GA3 did not affect the expression of these genes. In bolting plants, both ERD10 and ERD14 transcripts were detected in stems, cauline leaves, roots and flowers, but their levels decreased during seed formation and only low levels of the transcripts were detected in seeds at the late-mature stage.

Amino Acid Sequence↗

The CRK1 receptor-like kinase gene of tobacco is negatively regulated by cytokinin.

This report describes a novel receptor-like kinase gene of tobacco (Nicotiana tabacum L.) that, in cell culture, is rapidly regulated by very low concentrations of cytokinin. The steady-state transcript level of the CYTOKONIN-REGULATED KINASE 1 gene (CRK1) was strongly reduced 30 min after cytokinin treatment. At higher concentrations abscisic acid and auxin induced a similar response. None of the other plant hormones tested elicited this response. Further analyses of the cytokinin-dependentregulation showed that the reduction of transcript was transient, and the duration of the recovery phase was dependent on the hormone concentration. CRKI is not a primary response gene as the simultaneous addition of cycloheximide inhibits its regulation by cytokinin. Inhibitor studies revealed that a protein phosphatase is likely involved in signalling processes upstream of CRK1. CRKI is expressed at low levels in the leaves, stem and roots of tobacco. It is predicted that the CRK1 protein is located in the plasma membrane. It has in its N-terminal putative receptor sequence a signal peptide, a serine- and a proline-rich region, a six repeat motif similar to the CRINKLY4 protein of Zea mays and several regions homologous to purine-binding motifs. A single transmembrane domain is followed by a highly conserved intracellular Ser/Thr kinase domain. Therefore, CRKI is a novel type of class I plant receptor kinase. We hypothesize that CRKI is involved in an early step of hormone signalling and that transcript down-regulation reflects a desensitization step in reaction to the signalling molecule.

Base Sequence↗

Increased gibberellin biosynthesis in transgenic trees promotes growth, biomass production and xylem fiber length.

In most tree-breeding programs worldwide, increasing the trees' growth rates and stem volumes and shortening their rotation times are important aims. Such trees would yield more biomass per unit area. Here we show that overexpressing a key regulatory gene in the biosynthesis of the plant hormone gibberellin (GA) in hybrid aspen (Populus tremula x P. tremuloides) improves growth rate and biomass. In addition, these transgenic trees have more numerous and longer xylem fibers than unmodified wild-type (wt) plants. Long fibers are desirable in the production of strong paper, but it has not as yet proved possible to influence this trait by traditional breeding techniques. We also show that GA has an antagonistic effect on root initiation, as the transgenic lines showed poorer rooting than the control plants when potted in soil. However, the negative effect on rooting efficiencies in the initial establishment of young plantlets in the growth chamber did not significantly affect root growth at later stages.

Blotting, Northern↗