Showing papers in "Journal of Plant Growth Regulation in 2007"

Induction of Abiotic Stress Tolerance by Salicylic Acid Signaling

Abstract: The role of salicylic acid (SA) as a key molecule in the signal transduction pathway of biotic stress responses has already been well described. Recent studies indicate that it also participates in the signaling of abiotic stresses. The application of exogenous SA could provide protection against several types of stresses such as high or low temperature, heavy metals, and so on. Although SA may also cause oxidative stress to plants, partially through the accumulation of hydrogen peroxide, the results published so far show that the preliminary treatment of plants with low concentrations of SA might have an acclimation-like effect, causing enhanced tolerance toward most kinds of abiotic stresses due primarily to enhanced antioxidative capacity. The effect of exogenous SA depends on numerous factors such as the species and developmental stage of the plant, the mode of application, and the concentration of SA and its endogenous level in the given plant. Recent results show that not only does exogenous SA application moderate stress effects, but abiotic stress factors may also alter the endogenous SA levels in the plant cells. This review compares the roles of SA during different abiotic stresses.

Ethylene and Fruit Ripening

Abstract: The ripening of fleshy fruits represents the unique coordination of developmental and biochemical pathways leading to changes in color, texture, aroma, and nutritional quality of mature seed-bearing plant organs. The gaseous plant hormone ethylene plays a key regulatory role in ripening of many fruits, including some representing important contributors of nutrition and fiber to the diets of humans. Examples include banana, apple, pear, most stone fruits, melons, squash, and tomato. Molecular exploration of the role of ethylene in fruit ripening has led to the affirmation that mechanisms of ethylene perception and response defined in the model system Arabidopsis thaliana are largely conserved in fruit crop species, although sometimes with modifications in gene family size and regulation. Positional cloning of genes defined by ripening defect mutations in the model fruit system tomato have recently led to the identification of both novel components of ethylene signal transduction and unique transcription factor functions influencing ripening-related ethylene production. Here we summarize recent developments in the regulation of fruit ripening with an emphasis on the regulation of ethylene synthesis, perception, and response.

Regulation of Ethylene Biosynthesis

Abstract: The biosynthesis of the gaseous phytohormone ethylene is a highly regulated process A major point of regulation occurs at the generally rate-limiting step in biosynthesis, catalyzed by the enzyme ACC synthase (ACS) ACS is encoded by a multigene family, and different members show distinct patterns of expression during growth and development, and in response to various external cues In addition to this transcriptional control, the stability of the ACS protein is also highly regulated Here we review these two distinct regulatory inputs that control the spatial and temporal patterns of ethylene biosynthesis

Deciphering the role of ethylene in plant-herbivore interactions

Abstract: Most plants emit ethylene in response to herbivory by insects from many different feeding guilds. The elicitors of these ethylene emissions are thought to be microorganisms or oral secretion-specific compounds that are transferred when the attacking insect feeds. To find the receptors for these elicitors and describe the signaling cascades that are subsequently activated will be the challenge of future research. Past experiments on the function of herbivore-induced ethylene, which were biased toward the use of chemical treatments to manipulate ethylene, identified seven ethylene-dependent defense responses. In contrast, a genetic toolbox that consists of several mutants has rarely been used and to date, mutants have helped to identify only one additional ethylene-dependent defense response. Ethylene-dependent responses include the emission of specific volatile organic compounds as indirect defense, the accumulation of phenolic compounds, and proteinase inhibitor activity. Besides being ethylene regulated, these defenses depend strongly on the wound-hormone jasmonic acid (JA). That ethylene requires the concomitant induction of JA, or other signals, appears to be decisive. Rather than being the principal elicitor of defense responses, ethylene modulates the sensitivity to a second signal and its downstream responses. Given this modulator role, and the artifacts associated with the use of chemical treatments to manipulate ethylene production and perception, future advances in the study of ethylene’s function in plant–herbivore interactions will likely come from the use of signaling mutants or transgenic plants. It will be exciting to see if adaptive phenotypic plasticity is largely an ethylene-mediated response.

Effect of Gibberellin and Auxin on Parthenocarpic Fruit Growth Induction in the cv Micro-Tom of Tomato

Abstract: The effect of applied gibberellin (GA) and auxin on fruit-set and growth has been investigated in tomato (Solanum lycopersicum L.) cv Micro-Tom. It was found that to prevent competition between developing fruits only one fruit per truss should be left on the plant. Unpollinated ovaries responded to GA3 and to different auxins [indol-3-acetic acid, naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid (2,4-D)], 2,4-D being the most efficient. GA3- and 2,4-D-induced fruits had different internal morphology, with poor locular tissue development in the case of GA, and pseudoembryos development in the case of 2,4-D. Also, GA3 produced larger cells in the internal region of the mesocarp (IM) associated with higher mean C values, whereas 2,4-D produced more cell layers in the pericarp than pollinated fruits. The smaller size of GA3- compared with 2,4-D-induced fruits was due to them having fewer cells, only partially compensated by the larger size of IM cells. Simultaneous application of GA3 and 2,4-D produced parthenocarpic fruits similar to pollinated fruits, but for the absence of seeds, suggesting that both kinds of hormones are involved in the induction of fruit development upon pollination. It is concluded that Micro-Tom constitutes a convenient model system, compared to tall cultivars, to investigate the hormonal regulation of fruit development in tomato.

Biochemical Characterization of Cytokinin Oxidases/Dehydrogenases from Arabidopsis thaliana Expressed in Nicotiana tabacum L.

Abstract: Transgenic tobacco plants overexpressing single Arabidopsis thaliana cytokinin dehydrogenase (CKX, EC 1.5.99.12) genes AtCKX1, AtCKX2, AtCKX3, AtCKX4, AtCKX5, AtCKX6, and AtCKX7 under the control of a constitutive 35S promoter were tested for CKX-enzymatic activity with varying pH, electron acceptors, and substrates. This comparative analysis showed that out of these, only AtCKX2 and AtCKX4 were highly active enzymes in reaction with isoprenoid cytokinins (N 6 -(2-isopentenyl)adenine (iP), zeatin (Z)) and their ribosides using the artificial electron acceptors 2,6-dichlorophenol indophenol (DCPIP) or 2,3-dimethoxy-5-methyl-1,4-benzoquinone (Q0). Turnover rates of these cytokinins by four other AtCKX isoforms (AtCKX1, AtCKX3, AtCKX5, and AtCKX7) were substantially lower, whereas activity of AtCKX6 was almost undetectable. The isoenzymes AtCKX1 and AtCKX7 showed significant preference for cytokinin glycosides, especially N 6 -(2-isopentenyl)adenine 9-glucoside, under weakly acidic conditions. All enzymes preferentially cleave isoprenoid cytokinins in the presence of an electron acceptor, but aromatic cytokinins are not resistant and are degraded with lower reaction rates as well. Cytokinin nucleotides, considered as resistant to CKX attack until now, were found to be potent substrates for some of the CKX isoforms. Substrate specificity of AtCKXs is discussed in this study with respect to the structure of the CKX active site. Further biochemical characterization of the AtCKX1, AtCKX2, AtCKX4 and AtCKX7 enzymes showed pH-dependent activity profiles.

Modulation of Plant Defenses by Ethylene

Abstract: Ethylene (ET) plays a critical role in the activation of plant defenses against different biotic stresses through its participation in a complex signaling network that includes jasmonic acid (JA), salicylic acid (SA), and abscisic acid (ABA). Pathogen attack, wounding, and herbivory trigger asymmetric activation of this defense signaling network, thereby affecting the final balance of interactions between its components and establishing a targeted response to the initial threat. Ethylene’s contribution to the modulation of this defense network relies on the complexity of the regulation of multigene families involved in ET biosynthesis, signal transduction, and crosstalk and enables the plant to fine-tune its response. The function of the members of these multigene families is tightly regulated at transcriptional, post-transcriptional, and post-translational levels. It is generally accepted that ET cooperates with JA in the activation of defenses against necrotrophic pathogens and antagonizes SA-dependent resistance against biotrophic pathogens. However, this is likely an oversimplified view, because cooperative interactions between ET and SA pathways have been reported and ET has been implicated in the activation of defenses against some biotrophic and hemibiotrophic pathogens. Therefore, deciphering ET’s place in this hormonal network is essential to understanding how the cell orchestrates an optimal response to a specific biotic stress.

Growth Control by Ethylene: Adjusting Phenotypes to the Environment

Abstract: Plants phenotypically adjust to environmental challenges, and the gaseous plant hormone ethylene modulates many of these growth adjustments. Ethylene can be involved in environmentally induced growth inhibition as well as growth stimulation. Still, ethylene has long been considered a growth inhibitory hormone. There is, however, accumulating evidence indicating that growth promotion is a common feature in ethylene responses. This is evident in environmental challenges, such as flooding and competition, where the resulting avoidance responses can help plants avoid adversity. To show how ethylene-mediated growth enhancement can facilitate plant performance under adverse conditions, we explored a number of these examples. To escape adversity, plants can optimize growth and thereby tolerate abiotic stresses such as drought, and this response can also involve ethylene. In this article we indicate how opposing effects of ethylene on plant growth can be brought about, by discussing a unifying, biphasic ethylene response model. To understand the mechanistic basis for this multitude of ethylene-mediated growth responses, the involvement of ethylene in processes that control cell expansion is also reviewed.

Exogenous Abscisic Acid Increases Carbohydrate Accumulation and Redistribution to the Grains in Wheat Grown Under Field Conditions of Soil Water Restriction

Abstract: This work investigates the effects of abscisic acid (ABA) on physiologic parameters related to yield in wheat (Triticum aestivum) grown under field conditions with water restriction ranging between 457% and 495% of field capacity during anthesis and postanthesis ABA (300 mg L−1) was sprayed onto the plants at the beginning of shoot lengthening which significantly promoted leaf area and higher concentrations of chlorophylls and carotenoids in flag leaf at anthesis ABA also increased soluble carbohydrates in shoots at anthesis, which were then re-exported to the grains at maturity This correlated with a yield increase that was achieved by a higher number and weight of grains per spike, but protein content was not significantly affected

Ethylene Receptors: Ethylene Perception and Signal Transduction

Abstract: Ethylene is sensed by a family of receptors that can be divided into two subfamilies based on phylogenetic analysis and some shared structural features. In this review we focus on the mechanistic aspects of how the receptors function in plants to transduce the ethylene signal. Recent work has led to new insights into how ethylene binds to the receptors and how this binding may induce a conformational change to regulate signaling. Additional studies point to several possible mechanisms for signal output by the receptors, which may involve changes in enzymatic activity and/or conformational changes. Other studies indicate the importance of interactions, both physical and genetic, between the receptors and early components of the signaling pathway, in particular, the Raf-like kinase CTR1, which functions as an integral component of the ethylene receptor signaling complex. The current model for signaling in Arabidopsis supports differing contributions from the receptors, with subfamily-1 receptors playing a more significant role than the subfamily-2 receptors in transmitting the ethylene signal.

New Techniques for the Estimation of Naturally Occurring Brassinosteroids

Abstract: We have developed enzyme-linked immunosorbent assays (ELISAs) for measuring 24-epicastasterone and related brassinolide analogs, with detection ranges of 0.005 to 50 pmoles. Polyclonal antibodies used in these assays were raised against 24-epicastasterone carboxymethyloxime-bovine serum albumin conjugates and were found to have high specificity for 24-epibrassinosteroids. Natural brassinosteroids (BRs), such as brassinolide and 24-epibrassinolide, exhibited relatively high cross-reactivities with the generated antibodies, whereas other BR analogs with β-oriented hydroxyl groups at C-2, C-3, C-22, and C23 lacked immunoreactivity. Through the use of internal standardization, dilution assays, recovery of authentic [3H]24-epicastasterone, and immunohistograms, the ELISAs have been shown to be applicable for estimating 24-epibrassinosteroid levels in crude plant extracts. To analyze brassinosteroids in tissues from young bean (Phaseolus vulgaris L., cv. Pinto), Daucus carota ssp.sativus plants and Arabidopsis thaliana L. Heynh. seedlings, and rape (Brassica napus L.) pollen, the extracts were fractionated by high performance liquid chromatography (HPLC) and the resulting fractions were analyzed by the ELISA method. Immunohistogram ELISA analysis of HPLC fractions indicated that major peaks of immunoreactivity co-chromatographed with the labeled and unlabeled 24-epibrassinolide. A highly sensitive electrospray ionization mass spectrometry (MS) technique (LOD: 50 fmol) was also developed and the results obtained by the HPLC-ELISA and HPLC-MS approaches were compared.

Cloning and Biochemical Characterization of ToFZY, a Tomato Gene Encoding a Flavin Monooxygenase Involved in a Tryptophan-dependent Auxin Biosynthesis Pathway

Abstract: Indole-3-acetic acid (IAA), the main endogenous auxin, has been known for decades to be a key regulator for plant growth and development. Multiple routes have been proposed for IAA biosynthesis but physiologic roles or relevance of the different routes are still unclear. Recently, four members of the Arabidopsis thaliana YUC gene family have been implicated in an additional requirement of IAA involved in floral organ and vascular tissue formation. The loss-of-function yuc1yuc4 double mutants in Arabidopsis displayed phenotypes similar to the previously described loss-of-function floozy mutants in petunia (fzy). Moreover, it has been demonstrated that YUC1 encodes a flavin monooxygenase (FMO) that catalyzes a rate-limiting step of a tryptophan-dependent auxin biosynthesis pathway: the conversion of tryptamine to N-hydroxyl-tryptamine. Here we report on the genetic study of ToFZY, the putative tomato ortholog of YUC4 and FZY, including gene and cDNA sequence comparison and a preliminary expression analysis. In addition, we describe a novel conserved amino acid motif that may be considered a hallmark potentially useful for the identification of new YUC-like FMOs. We also demonstrate that ToFZY encodes a protein with the same enzymatic activity as YUC1. Finally, we provide evidence suggesting that the ToFZY gene belongs to a multigenic family whose members may exhibit a temporal and spatial specialization similar to that described in A. thaliana.

Abscisic Acid and Ethylene Interact in Rice Spikelets in Response to Water Stress During Meiosis

Abstract: This study investigated whether and how the interaction between abscisic acid (ABA) and ethylene is involved in the regulation of rice (Oryza sativa L.) spikelet sterility when subjected to water stress during meiosis. Two rice cultivars, HA-3 (drought-resistant) and WY-7 (drought-susceptible), were used and subjected to well-watered and water-stressed (WS) treatments during meiosis (15–2 days before heading). Leaf water potentials of both cultivars markedly decreased during the day as a result of the WS treatment, but panicle water potentials remained constant. The percentage of sterile spikelets in WS plants was increased by 49.7% for WJ-7 but only 12.7% for HA-3. ABA, ethylene, and 1-aminocyclopropane-1-carboxylic acid were all enhanced in spikelets by the water stress, but ethylene was enhanced more than ABA in WY-7 when compared with that in HA-3. Spikelet sterility was significantly reduced when ABA or amino-ethoxyvinylglycine, an inhibitor of ethylene synthesis, was applied to the panicles of WS plants at the early meiosis stage. Application of ethephon, an ethylene-releasing agent, or fluridone, an inhibitor of ABA synthesis, had the opposite effect, and sterility was increased. The results suggest that antagonistic interactions between ABA and ethylene may be involved in mediating the effect of water stress on spikelet fertility. A higher ratio of ABA to ethylene would be a physiologic trait of rice adaptation to water stress.

Molecular Basis of the Ethylene Signaling and Response Pathway in Arabidopsis

Abstract: The gaseous phytohormone ethylene is a key regulator in plant growth and developmental process as well as biotic and abiotic stress response. This review focuses on the recent advances in the ethylene-signaling pathway in Arabidopsis, with particular emphasis on the latest information about the downstream events of the ethylene-response pathway. Notable new findings include identification of a specific regulator of the ethylene receptor ETR1, discovery of protein degradation and RNA turnover processes in modulating EIN3-dependent transcriptional regulation, demonstration of the involvement of auxin biosynthesis in ethylene-mediated inhibition of root growth, and determination of possible integration points between ethylene and other hormonal and environmental signals (gibberellin, jasmonic acid, light, and sugar) in various plant processes. The elucidation of the molecular mechanisms of the ethylene-signaling and ethylene- response pathway in Arabidopsis might provide a framework for understanding how other plant species sense and respond to ethylene.

The Effect of Plant Growth Regulators and Sucrose on the Micropropagation and Microtuberization of Dioscorea nipponica Makino

Abstract: The effects of sucrose, plant growth regulators, MS (Murashige and Skoog), and ½MS salt media formulations were investigated for the development of shoot cultures, microtuber induction, and plantlet regeneration in Dioscorea nipponica. The cytokinin N-benzyladenine (BA) in the range of 0.5–2.0 mg/l showed strong enhancing effects on microtuber induction only when used in conjunction with the auxin alpha-naphthalene acetic acid (NAA), with the effect that NAA increased from 0.5 to 2.0 mg/l. Murashige and Skoog salt media supplemented with sucrose at 3% (w/v) gave the highest frequencies of shoot induction (86%) when BA was present at 2.0 mg/l and NAA at 1.0 mg/l. Sucrose at 7% (w/v) was the single most significant medium constituent for microtuber growth. The heaviest microtubers were formed on media containing 1.0 mg/l BA and 2.0 mg/l (0.073 g), especially with 7% sucrose (3.46 g). With media containing ½MS, 2% sucrose, and 0.1% (w/v) activated charcoal, the percentage of rooting was maximal when supplemented with 1.0 mg/l BA and 0.5 mg/l NAA for the in vitro produced shoots (95%) and BA and NAA both at 0.5 mg/l for the microtubers (100%). When removed from culture flasks and transferred into sterilized soil in a greenhouse, most of the hardened plantlets survived (over 91% after 1 week), and they were suitable for field planting after 1 month.

Small GTP-binding Proteins and their Functions in Plants

Abstract: Small GTP-binding proteins exist in eukaryotes from yeast to animals to plants and constitute a superfamily whose members function as molecular switches that cycle between “active” and “inactive” states. They regulate a wide variety of cell functions such as signal transduction, cell proliferation, cytoskeletal organization, intracellular membrane trafficking, and gene expression. In yeast and animals, this superfamily is structurally classified into at least five families: the Ras, Rho, Rab, Arf/Sar1, and Ran families. However, plants contain Rab, Rho, Arf, and Ran homologs, but no Ras. Small GTP-binding proteins have become an intensively studied group of regulators not only in yeast and animals but also in plants in recent years. In this article we briefly review the class and structure of small GTP-binding proteins. Their working modes and functions in animals and yeast are listed, and the functions of individual members of these families in plants are discussed, with the emphasis on the recently revealed plant-specific roles of these proteins, including their cross-talk with plant hormones and other signals, regulation of organogenesis (leaf, root, and embryo), polar growth, cell division, and involvement in various stress and defense responses.

Disrupting Abscisic Acid Homeostasis in Western White Pine (Pinus monticola Dougl. Ex D. Don) Seeds Induces Dormancy Termination and Changes in Abscisic Acid Catabolites

Abstract: To investigate the role of abscisic acid (ABA) biosynthesis and catabolism in dormant imbibed seeds of western white pine (Pinus monticola), ABA and selected catabolites were measured during a combined treatment of the ABA biosynthesis inhibitor fluridone, and gibberellic acid (GA). Fluridone in combination with GA effectively disrupted ABA homeostasis and replaced the approximately 90-day moist chilling period normally required to break dormancy in this species. Individually, both fluridone and GA treatments decreased ABA levels in the embryos and megagametophytes of white pine seeds compared to a water control; however, combined fluridone/GA treatment, the only treatment to terminate dormancy effectively, led to the greatest decline in ABA content. Fluridone treatments revealed that a high degree of ABA turnover/transport occurred in western white pine seeds during the initial stages of dormancy maintenance; at this time, ABA levels decreased by approximately two-thirds in both embryo and megagametophyte tissues. Gibberellic acid treatments, both alone and in combination with fluridone, suggested that GA acted transiently to disrupt ABA homeostasis by shifting the ratio between biosynthesis and catabolism to favor ABA catabolism or transport. Increases in phaseic acid (PA) and dihydrophaseic acid (DPA) were observed during fluridone/GA treatments; however, increases in ABA metabolites did not account for the reduction in ABA observed; additional catabolism and/or transport of ABA and selected metabolites in all probability accounts for this discrepancy. Finally, levels of 7′ hydroxy-ABA (7′OH-ABA) were higher in dormant-imbibed seeds, suggesting that metabolism through this pathway is increased in seeds that maintain higher levels of ABA, perhaps as a means to further regulate ABA homeostasis.

Plant Growth Regulators and Induction of Leaf Senescence in Nitrogen-Deprived Wheat Plants

Abstract: The sequence of events and the signals that regulate the remobilization of nitrogen (N) reserves during senescence induced by N starvation were studied in leaf 3, the last fully expanded leaf, in 17-day-old wheat (Triticum aestivum L.) plants. The first event observed was a rapid decrease in the isopentenyl adenosine (iPA) concentration during the first 24 h of N starvation. No differences in t-zeatin riboside and dihydrozeatin riboside concentrations were observed until the end of the assay. During the following 6 days, a decrease in soluble amino acids, chlorophyll, and protein, as well as an increase in soluble sugar concentration and endoproteolytic activity, could be observed. At day 3 of the experiment, the abscisic acid (ABA) concentration in the leaves of N-deprived plants started to increase. After 6 days of N deprivation there was a rise in oxidative stress, as indicated by the increase in malondialdehyde concentration, as well as a decrease in the activities of antioxidant enzymes catalase and ascorbate peroxidase. To analyze interactions with leaf development, the first, second, third, and fourth leaves were studied. iPA concentration decreased in all the leaf stages, including leaf 4, which was not fully expanded. A linear correlation between iPA and protein concentration was determined. These results suggest that the sharp fall in iPA could be the earliest event that induces protein degradation during the development of senescence induced by N deficiency, and that only later is ABA accumulated and oxidative stress developed.

One for All and All for One: Cross-Talk of Multiple Signals Controlling the Plant Phenotype

Abstract: The plant hormone ethylene plays a pivotal role in steering various processes by regulating the biosynthesis, distribution, or signal transduction of other hormones. Ethylene also mediates the effects of other hormones. Similarly, hormones control the ethylene synthesis and signalling pathway. Eventually, integration of this network of signals leads to an appropriate morphological or biochemical response. Consequently, this cross-talk results in the characteristic plasticity associated with plant development. Here, the interplay of ethylene with other hormones is described for germination and seedling growth, stomatal control, and tissue elongation. The mechanisms by which this occurs are discussed in more detail.

Hormone Profiling by LC-QToF-MS/MS in Dormant Macadamia integrifolia : Correlations with Abnormal Vertical Growth

Abstract: A method for analyzing multiple plant hormone groups in small samples with a complex matrix was developed to initiate a study of the physiology of abnormal vertical growth (AVG) in Macadamia integrifolia (cv. HAES344). Cytokinins (CKs), gibberellins (GAs), abscisic acid (ABA), and auxins were detected in xylem sap and apical and lateral buds using high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QToF-MS/MS). The extraction method separated compounds with high sensitivity in positive (CKs) and negative (ABA, auxins, GAs) modes of QToF-MS/MS. CK profiles differed in xylem sap and apical and lateral buds irrespective of AVG symptoms. Trans-zeatin riboside (t-ZR) was dominant in sap of normal and AVG trees (∼4 and 6 pmol g−1 FW, respectively). In apical buds isopentenyl adenine (iP) (∼30 pmol g−1 FW) was the most abundant CK, and in lateral buds trans-zeatin (t-Z) (22–24 pmol g−1 FW) and iP (24–30 pmol g−1 FW) were the most abundant. t-Z levels of AVG trees were higher in apical buds (13.88 vs. 6.6 pmol g−1 FW, p < 0.05) and lower in sap (0.16 vs. 0.51 pmol ml−1, p < 0.005) compared to normal trees. ABA in lateral buds was 1.9 times higher (p < 0.001) in AVG. IAA was below quantification, whereas indole-3-butyric acid (IBA) was consistently present. GA7 was the dominant GA in apical and lateral buds of all trees (100–150 pmol g−1 FW). GA3, 4, & 9 were consistently present at low concentrations (<12 pmol g−1 FW) in buds. GAs1, 3, & 9 were detected in xylem sap at low concentrations (<0.5 pmol g−1 FW). Differences in sap amino acids (AA) were also assessed. In sap from AVG trees, asparagine and glutamine increased significantly (p < 0.05) in their contribution to total AA. Potential AVG hormone correlations are discussed.

A Crucial Role for Cytokinins in Pea ABR17-mediated Enhanced Germination and Early Seedling Growth of Arabidopsis thaliana under Saline and Low-temperature Stresses

Abstract: The role of cytokinins (CKs) in mediating the previously observed ABR17-mediated enhancement of germination of Arabidopsis thaliana under salinity and low-temperature stresses has been evaluated. We determined the endogenous concentrations of CK in the three transgenic and wild-type seedlings, which indicated that the transgenic seedlings had higher endogenous concentrations of CK. Furthermore, the relative levels of expression of ABR17 cDNA and the primary CK response gene, ARR5, were evaluated in the transgenic and wild-type seedlings by quantitative real-time polymerase chain reaction (RT-PCR). Our results indicated that two of the three independently derived transgenic plants possessed higher levels of ABR17 transcripts, which correlated well with increased ARR5 expression, further supporting a possible role for CK in mediating the observed phenomenon. In addition, the exogenous application of various CKs on the germination of wild-type A. thaliana under these abiotic stress conditions enhanced its germination. Finally, the ribonuclease (RNase) activity of the pea ABR17 protein was also demonstrated after expression of its cDNA in Escherichia coli, purification of the recombinant protein, and in vitro RNase assays. Our findings are discussed within the context of ABR17-mediated enhancement of endogenous CK concentrations, the involvement of CKs in germination under abiotic stress, as well as the role of the RNase activity of ABR17 protein in mediating the observed effects.

Responses of Chrysanthemum Cells to Mechanical Stimulation Require Intact Microtubules and Plasma Membrane–Cell Wall Adhesion

Abstract: Plant cells are highly susceptible and receptive to physical factors, both in nature and under experimental conditions. Exposure to mechanical forces dramatically results in morphological and microstructural alterations in their growth. In the present study, cells from chrysanthemum (Dendranthema morifolium) were subjected to constant pressure from an agarose matrix, which surrounded and immobilized the cells to form a cell-gel block. Cells in the mechanically loaded blocks elongated and divided, with an axis preferentially perpendicular to the direction of principal stress vectors. After a sucrose-induced plasmolysis, application of peptides containing an RGD motif, which interferes with plasma membrane-cell wall adhesion, reduced the oriented growth under stress conditions. Moreover, colchicines, but not cytochalasin B, abolished the effects of mechanical stress on cell morphology. Cellulose staining revealed that mechanical force reinforces the architecture of cell walls and application of mechanical force, and RGD peptides caused aggregative staining on the surface of plasmolyzed protoplasts. These results provide evidence that the oriented cell growth in response to compressive stress requires the maintenance of plasmalemma-cell wall adhesion and intact microtubules. Stress-triggered wall development in individual plant cells was also demonstrated.

Factors Affecting Somatic Embryogenesis Induction and Conversion in “Paradise Tree” (Melia azedarach L.)

Abstract: Factors affecting somatic embryogenesis induction and conversion in paradise tree (Melia azedarach) were evaluated. Somatic embryogenesis was influenced by plant growth regulators, explant stage, carbohydrate source and concentration, gelling agents, light, and induction times. MS medium with 4.54 μM thidiazuron (TDZ) was optimal for the induction of embryogenic tissue. Zygotic embryos that were 1-1.5 mm long (torpedo and early cotyledonal stage) had a greater embryogenic response than smaller or larger embryos and better conversion of somatic embryos into plants. In general, embryos that formed in medium containing 1% or 5% carbohydrate were hyperhydrics or fused, respectively, whereas those that formed in medium with a carbohydrate concentration of 3% had better morphology. Raffinose at 3% yielded satisfactory somatic embryo induction with good morphology and the best values of conversion into plants. Induction and conversion of somatic embryos were superior on medium solidified with agar A-1296. The explants maintained under 160 μmol m−2 s−1 or 1 week in darkness and later 160 μmol m−2 s−1 produced a significantly higher embryogenic index. Only 4 days of treatment on induction medium, with either raffinose or sucrose at 3% as a carbohydrate source, were required to induce somatic embryogenesis, but longer exposure, until 18 days, increased the yield and improved the morphology of somatic embryos.

Role of cAMP in Gibberellin Promotion of Seed Germination in Orobanche minor Smith

Abstract: Adenosine 3′,5′-cyclic monophosphate (cAMP) is known as a key second messenger in many living organisms, regulating a wide range of cellular responses. In higher plants the function of cAMP is poorly understood. In this study, we examined the role of cAMP in seed germination of the root parasitic plant Orobanche minor whose seeds require preincubation in warm moist environments for several days, termed conditioning, prior to exposure to germination stimulants released from roots of host plants. Accumulation of endogenous cAMP was observed in the conditioned O. minor seeds. When the seeds were exposed to light or supraoptimal temperature during the conditioning period, cAMP did not accumulate and the seeds showed low germination rates after stimulation with strigol, a germination stimulant. Addition of membrane-permeable cAMP to the medium restored the germination rates of the seeds treated with light or supraoptimal temperature during the conditioning period, suggesting that cAMP functions during the conditioning period. The endogenous cAMP levels of the seeds conditioned in the light or at a supraoptimal temperature were elevated by treatment of the seeds with gibberellin (GA) during the conditioning period. Uniconazole, a potent inhibitor of GA biosynthesis, blocked elevation of the cAMP level. Furthermore, a correlation between the endogenous cAMP level and GA level was observed during the conditioning period. These results suggest that GAs elevate the cAMP level, which is required for the germination of O. minor seeds.

A Bacterial Indole-3-acetyl-L-aspartic Acid Hydrolase Inhibits Mung Bean ( Vigna radiata L.) Seed Germination Through Arginine-rich Intracellular Delivery

Abstract: Indole-3-acetyl-L-aspartic acid (IAA-Asp) is a natural product in many plant species and plays many important roles in auxin metabolism and plant physiology. IAA-Asp hydrolysis activity is, therefore, believed to affect plant physiology through changes in IAA metabolism in plants. We applied a newly discovered technique, arginine-rich intracellular delivery (AID), to deliver a bacterial IAA-Asp hydrolase into cells of mung bean (Vigna radiata) seeds and measured its effects on mung bean seed germination. IAA-Asp hydrolase inhibited seed germination about 12 h after the enzyme was delivered into cells of mung bean seeds both covalently and noncovalently. Mung bean seed germination was delayed by 36 h when the enzyme protein was noncovalently attached to the AID peptide and longer than 60 h when the enzyme protein was covalently attached to the AID peptide. Root elongation of mung bean plants was inhibited as much as 90% or 80%, respectively, when the IAA-Asp hydrolase was delivered with the AID peptide by covalent or noncovalent association. Further thin-layer chromatography analysis of plant extracts indicated that the levels of IAA increased about 12 h after treatment and reached their peak at 24 h. This result suggests that IAA-Asp hydrolase may increase IAA levels and inhibit seed germination of mung bean plants and that the AID peptide is a new, rapid, and efficient experimental tool to study the in vivo activity of enzymes of interest in plant cells.

Differential Effect of Aluminum on DNA Synthesis and CDKA Activity in Two Coffea arabica Cell Lines

Abstract: Aluminum (Al) is widely known to inhibit root growth: however, the exact mechanism of root growth inhibition is uncertain. To address the effect of Al on some components of the culture cycle, we used two Coffea arabica cell lines, L2 (Al-sensitive) and LAMt (Al-tolerant), and evaluated [3H]-thymidine incorporation into DNA and cyclin dependent kinase type A (CDKA) activity. We used cell cultures of the 7th day that were incubated with AlCl3 (100 μM) during different periods of time and observed a decrease of 47% in the rate of [3H]-thymidine incorporation into DNA when the cells were incubated in the presence of Al in the L2 cell line as compared with the LAMt cell line, which showed an increase of 150% over the control. A 1,132 bp cDNA for CDKA was amplified from C. arabica cells using real-time polymerase chain reaction (RT-PCR) and 3′-RACE, which encoded a 294 amino acid polypeptide. Incubation of the cells with AlCl3 (100 μM) causes inhibition of CDKA activity up to 72% in the L2 cell line but it was stimulated up to 70% in the LAMt cell line. The Al effect on expression of CDKA transcripts estimated by RT-PCR showed no effect upon AlCl3 incubation between the two cell lines. Our results suggest that Al affects cell cycle elements differentially in the two tested cell lines.

Plant Hormones Isolated from ''Katahdin'' Potato Plant Tissues and the Influence of Photoperiod and Temperature on Their Levels in Relation to Tuber Induction

Abstract: Qualitative and quantitative analyses were carried out on vegetative tissues of potato (Solanum tuberosum cv. “Katahdin”) in search of natural products thought to play a role in tuber induction. Tissues were obtained from plants initially grown in a growth chamber under noninducing conditions (30°C day and 28°C night with an 18-h photoperiod), and then half of the plants were moved to inducing chambers (28°C day and 13°C night with a 10-h photoperiod) for 10 days prior to tissue harvest. Plants from each chamber were then harvested at 2-day intervals for 10 days, separated into above- and belowground portions, and the lyophilized tissues were extracted and subjected to rigorous purification and separation using high-performance liquid chromatography. This was followed by identification and quantification using combined gas chromatography-mass spectrometry. Compounds isolated and identified included gibberellic acid; cytokinins cis-zeatin riboside, trans-zeatin, trans-zeatin riboside, and isopentenyladenine; and jasmonates jasmonic acid, tuberonic acid and its methyl ester, methyl 7-isocucurbate, and 9,10-dihydromethyljasmonate. Methyl 7-isocucurbate and 9,10-dihydromethyljasmonate were detected for the first time in potato tissue as endogenous compounds. Cytokinin and jasmonate levels generally increased under inducing conditions, whereas gibberellic acid levels declined progressively during the 10-day sampling period. Only gibberellic acid, jasmonic acid, and cis-zeatin riboside levels were significantly influenced by induction.

Effect of Temperature on Growth and Phototropism of Arabidopsis thaliana Seedlings

Abstract: Seedlings of Arabidopsis thaliana grown at 25°C responded to a change in growth temperature by changing their elongation rate within the next 150 min. Regardless of whether the new temperature was higher or lower than 25°C, the seedlings grew slower after the transfer at all tested temperatures. When the seedlings were grown for 2 days at 11.5°C, 17.9°C, and 23.5°C and then transferred to the range of temperatures between 4°C and 38°C they exhibited maximum elongation in the temperature range between 18°C and 23°C. The kinetics of first positive phototropism in seedlings transferred from 25°C to 15°C differed from the kinetics exhibited by seedlings transferred from 25°C to 28°C. At 15°C, measurable curvature began 40–50 min after the blue light (BL) pulse and no straightening was evident within 150 min after the BL pulse. Seedlings transferred to 28°C exhibited kinetics of phototropism similar to the phototropic response of plants maintained at 25°C except that straightening began slightly faster in the seedlings at 28°C. Based on these results, it is concluded that changes in temperature conditions affect both the elongation rate of seedlings and a first positive phototropism and that phototropic curvature and subsequent straightening are independently controlled.

Temperature and the Growth of Plant Cells

Abstract: In cell elongation, the juvenile cell vacuolates, takes up water, and expands by irreversible extension of the growth-limiting primary walls. This process was elaborated analytically by Lockhart in the mid-1960s. His growth equation does not, however, include the influence of the environmental temperature at which cell growth takes place. In this article we consider a phenomenological model including temperature in the equation of growth. Also, by introducing the possible influence of growth regulators treated here as external perturbations, linear and nonlinear solutions are found. A comparison of experimental and theoretical results permits qualitative and quantitative conclusions concerning change in the magnitude of the cell wall yielding coefficient Φ as a function of both time and temperature (with or without external perturbations), which has acquired reasonable values throughout.

Cytokinin Regulation of Gene Expression in the AHP Gene Family in Arabidopsis thaliana

Abstract: In higher plants histidine-aspartate phosphorelays are involved in hormone and stress signaling via a two-component system of signal transduction. In this system a histidine-containing phosphotransmitter (HPt) mediates signal transmission from a sensory histidine kinase to a response regulator, providing integration and/or branching of several different signaling pathways. Five genes encoding HPts, AHP1-5, have been identified in Arabidopsis. Histidine-aspartate phosphorelays involving HPts have been at least partly implicated in cytokinin signaling. We analyzed the regulation by cytokinins of AHP gene expression. We compared the effects on steady–state levels of AHP transcripts of a short-term treatment with an aromatic cytokinin and increase in endogenous isoprenoid cytokinin levels using an activable ipt system in 8-day-old Arabidopsis seedlings. Following ipt activation, a rapid and highly preferential increase in trans-zeatin-type cytokinins was observed, whereas other isoprenoid-type cytokinins showed no or only marginal increases. The levels of cytokinin metabolites under long-term ipt activation suggest that the seedlings may have difficulties in efficiently downregulating active forms of the hormone. Using real-time RT-PCR, transient increases in steady-state levels of AHP1-4 transcripts in response to both the short-term N 6-benzyladenine treatment and the increase in endogenous trans-zeatin-type cytokinin levels were observed. In contrast, both the full and the alternatively spliced AHP5 transcripts remained unaltered. On the other hand, increases in steady-state levels of AHP1-4 transcripts observed in seedlings cultivated continuously in the presence of exogenous N 6-benzyladenine were not paralleled in seedlings with constitutively increased endogenous trans-zeatin-type cytokinins, providing further indirect evidence for distinct functions of aromatic and isoprenoid cytokinins.