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Dual promoter of Agrobacterium tumefaciens mannopine synthase genes is regulated by plant growth hormones.

Temporal and spacial distribution of mannopine synthase (mas) promoter activity was determined throughout the development of transgenic tobacco plants using bacterial luciferase luxA and luxB as reporter genes. Luciferase activity was determined by luminometry in vitro and visualized by computer-enhanced single-photon video imaging in vivo. The activity of the mas dual promoters increased basipetally in developing plants and was wound-inducible in leaf and stem tissue. Hormone bioassays with isolated plant tissues and tumors deficient in the transferred DNA (T-DNA)-encoded genes iaaM, iaaH, and ipt indicated that activity of the mas dual promoters is regulated by auxin and enhanced by cytokinin in both differentiated and tumorous plant cells.

Journal Article↗

Comparative study of plant growth hormone (herbicide) toxicity in various biological subjects.

Due to their widespread distribution and toxic nature, herbicides may have a serious impact on the environment and exert adverse effects on associated organisms. The present study was conducted to determine the acute toxicological effects of some plant growth hormones used as herbicides on four biological subjects and compare the subjects' sensitivity to individual testing substances. The herbicides 4-(indol-3-yl)acetic acid C10H9O2N (IAA), N6-(beta 2-isopentenyl)adenosine (pi-indolylpropionic acid) C11H11O2N (IPA), 2,4-dichlorophenoxyacetic acid C8H6O3Cl2 (2,4-D), 4-chloro-2-methylphenoxyacetic acid C9H9O3Cl (MCPA), and 1,napthylacetic acid C12H10O2 (NAA) were tested and the following biological subjects were used: Daphnia magna, Tubifex tubifex, Scenedesmus quadricauda, and seeds of Sinapis alba. For S. alba, the influence of herbicides on seed germination (G) and root growth inhibition (I) was observed. For T. tubifex, the tests lasted 96 hr, for D. magna 48 hr, for S. quadricauda 20 days, and for S. alba 72 hr. The rank order of toxicity of herbicides used for T. tubifex was NAA > IAA > IPA > 2,4-D > MCPA; for D. magna. NAA > IAA > IPA > MCPA > 2,4-D; for S. quadricauda, IAA > IPA > NAA > MCPA > 2,4-D; for S. alba seed germination, NAA > IPA > 2,4-D > MCPA > IAA; and for root growth inhibition. NAA > 2,4-D > MCPA > IAA > IPA.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Further analogues of plant peptide hormone phytosulfokine-alpha (PSK-alpha) and their biological evaluation.

Phytosulfokine-alpha (PSK-alpha), a sulfated growth factor of structure H-Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-Gln-OH universally found in both monocotyledons and dicotyledons, strongly promotes proliferation of plant cells in culture. In studies on the structure/activity relationship of PSK-alpha the synthesis was performed of a series of a further 23 analogues modified in position 1, 3 or 4 as well as simultaneously in positions 1 and 3 of the peptide chain. Peptides were synthesized by the solid phase method according to the Fmoc procedure on a Wang-resin. Free peptides were released from the resin by 95% TFA in the presence of EDT. All peptides were tested by competitive binding assay to the carrot membrane using 3H-labelled PSK-alpha according to the test of Matsubayashi et al. Among these peptide analogues, [H-Phe(4-Cl)1]-PSK-alpha (IV), [H-Phe(4-I)1]-PSK-alpha (VII), and [Phe(4-Cl)3]-PSK-alpha (XI) retained 30% PSK-alpha activity. Analogue [Tyr(PO3H2)3]-PSK-alpha (IX) showed 10% of PSK-alpha activity.

Binding, Competitive↗

Plant peptide hormone phytosulfokine (PSK-alpha): synthesis of new analogues and their biological evaluation.

Phytosulfokine-alpha (PSK-alpha), a sulfated growth factor (H-Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-Gln-OH) universally found in both monocotyledons and dicotyledons, strongly promotes proliferation of plant cells in culture. In our studies on structure/activity relationship in PSK-alpha the synthesis of a series of analogues was performed: [H-D-Tyr(SO3H)1]- (9), [H-Phe(4-SO3H)1]- (10), [H-D-Phe(4-SO3H)1]- (11), [H-Phg(4-SO3H)1]- (12), [H-D-Phg(4-SO3H)1]- (13), H-Phe(4-NHSO2CH3)1]- (14), [H-D-Phe(4-NHSO2CH3)1]- (15), [H-Phe(4-NO2)1]- (16), [H-D-Phe(4-NO2)1]- (17), [H-Phg(4-NO2)1]- (18), [H-D-Phg(4-NO2)1]- (19), [H-Hph(4-NO2)1]- (20), [H-Phg(4-OSO3H)1]- (21), [Phe(4-NO2)3]- (22), [Phg(4-NO2)3]- (23), [Hph(4-NO2)3]- (24), [H-Phe(4-SO3H)1, Phe(4-SO3H)3]- (25) [H-Phe(4-NO2)1, Phe(4-NO2)3]- (26), [H-Phg(4-NO2)1, Phg(4-NO2)3]- (27), [H-Hph(4-NO2)1, Hph(4-NO2)3]- (28) and [Val3]- PSK-alpha (29). For modification of the PSK-alpha peptide chain the novel amino acids and their derivatives were synthesized, such as: H-L-Phg(4-SO3H)-OH (1), H-D-Phg(4-SO3H)-OH (2), Fmoc-Phg(4-SO3H)-OH (3), Fmoc-D-Phg(4-SO3H)-OH (4), Boc-Phg(4-NHSO2CH3)-OH (5), Boc-D-Phg(4-NHSO2CH3)-OH (6) Boc-Phe(4-NHSO2CH3)-OH (7), and Boc-D-Phe(4-NHSO2CH3)-OH (8). Peptides were synthesized by a solid phase method according to the Fmoc procedure on a Wang-resin. Free peptides were released from the resin by 95% TFA in the presence of EDT. All peptides were tested by competitive binding assay to the carrot membrane using 3H-labelled PSK according to the Matsubayashi et al. test.

Daucus carota↗

The plant wound hormone systemin binds with the N-terminal part to its receptor but needs the C-terminal part to activate it.

Suspension-cultured cells of Lycopersicon peruvianum respond with rapid medium alkalinization and a strong increase of a MAP kinase-like activity when treated with subnanomolar concentrations of the plant wound hormone systemin. Systemin fragments comprising the N-terminal 14 amino acids (syst1-14) or the C-terminal four amino acids (syst15-18), added singly or in combination, were inactive as inducers of these responses. Syst1-14 but not syst15-18 antagonized activity of intact systemin in a competitive manner. Likewise, intact systemin showed stimulatory, syst1-14 antagonistic activity, and syst15-18 showed no activity in leaf pieces of tomato (L. esculentum) plants assayed for the induction of ethylene biosynthesis. To study the molecular basis of perception, we extended the C-terminal end of systemin by a tyrosine residue and radioiodinated it to yield systemin-125I-iodotyrosine. In membrane preparations of L. peruvianum, this radioligand exhibited rapid, saturable, and reversible binding to a single class of binding sites. Binding showed a dissociation constant of approximately 1 nM, and binding of radioligand was efficiently competed by unlabeled systemin but not by syst15-18 or structurally unrelated peptides. Binding was also competed by the systemin antagonists syst1-14 and syst-Ala-17 (IC50 of 500 and 1000 nM, respectively). Thus, this binding site exhibits the characteristics expected for a functional systemin receptor. Based on these results, we propose a two-step mechanism for systemin action, with binding of the N-terminal part to the receptor as the first step and activation of responses with the C-terminal part as the second step.

Amino Acid Sequence↗

Marked changes in volume of mesophyll protoplasts of pea (Pisum sativum) on exposure to growth hormones.

The present study reports quick and significant changes induced by plant hormones in the volume of mesophyll protoplasts of pea (Pisum sativum). Four plant hormones: gibberellic acid (GA3), indole 3-acetic acid (IAA), abscisic acid (ABA)(+/-) and methyl jasmonate (MJ), caused marked changes in the volume of mesophyll protoplasts. GA3 and IAA increased the volume of the protoplasts (up to 90%) whereas the ABA and MJ decreased (by about 40%) the volume. Aquaporins or water channels appear to play an important role in swelling/shrinkage of the protoplasts as indicated by the suppression of volume changes by HgCl2 and reversal by mercaptoethanol. The possible role of secondary messengers in volume changes induced by GA3 was investigated by using selected pharmacological reagents. The GA3 induced swelling was restricted by GDP-beta-S (G-protein antagonist), U73122 (phospholipase C inhibitor), and TFP (calmodulin antagonist), but was not affected by 1-butanol (phospholipase D inhibitor), GTP-gamma-S (G-protein agonist), or verapamil (calcium channel blocker). The results suggest that the mesophyll protoplasts can be a simple and useful system for further studies on volume changes in plant tissues.

Abscisic Acid↗

The diversity of chemical substances controlling the nyctinastic leaf-movement in plants.

Leaf-movement in nyctinastic plants has long been believed to be controlled by plant hormones that are common among all nyctinastic plants. We have identified several bioactive substances for nyctinasty, whose bioactivities were highly specific to the original plant, based on the bioassay using the original plant leaf, and have shown that nyctinastic leaf-movement is not regulated by plant hormones. Our present results are in accordance with Umrath et al. physiologically significant opinion.

Biological Assay↗

Ethylene signal transduction in Arabidopsis.

It has been more than a centaury since the identification of ethylene as a gaseous plant hormone, and the effects of ethylene on plants have enabled broad applications in agriculture. Efforts had been made to understand ethylene signal transduction in plants, but it had not been successful until the identification and cloning of the first plant hormone receptor gene, ETR1, in Arabidopsis a decade ago. Comprehensive genetic studies allow the dissecting of the signaling pathway to reveal the components involved, which makes the studies of ethylene signal transduction become one the most-understood signal transduction pathways in plants. The identification of the signaling components and the genetic model of ethylene signal transduction pathway are described in this review.

Arabidopsis↗

Elevated CO2 induces physiological, biochemical and structural changes in leaves of Arabidopsis thaliana.

Leaves of Arabidopsis thaliana grown under elevated or ambient CO2 (700 or 370 micromol mol(-1), respectively) were examined for physiological, biochemical and structural changes. Stomatal characters, carbohydrate and mineral nutrient concentrations, leaf ultrastructure and plant hormone content were investigated using atomic absorption spectrophotometry, transmission electron microscopy and enzyme-linked immunosorbent assay (ELISA). Elevated CO2 reduced the stomatal density and stomatal index of leaves, and also reduced stomatal conductance and transpiration rate. Elevated CO2 increased chloroplast number, width and profile area, and starch grain size and number, but reduced the number of grana thylakoid membranes. Under elevated CO2, the concentrations of carbohydrates and plant hormones, with the exception of abscisic acid, increased whereas mineral nutrient concentrations declined. These results suggest that the changes in chloroplast ultrastructure may primarily be a consequence of increased starch accumulation. Accelerated A. thaliana growth and development in elevated CO2 could in part be attributed to increased foliar concentrations of plant hormones. The reductions in mineral nutrient concentrations may be a result of dilution by increased concentrations of carbohydrates and also of decreases in stomatal conductance and transpiration rate.

Arabidopsis↗

Transgenic studies on the involvement of cytokinin and gibberellin in male development.

Numerous plant hormones interact during plant growth and development. Elucidating the role of these various hormones on particular tissue types or developmental stages has been difficult with exogenous applications or constitutive expression studies. Therefore, we used tissue-specific promoters expressing CKX1 and gai, genes involved in oxidative cytokinin degradation and gibberellin (GA) signal transduction, respectively, to study the roles of cytokinin and GA in male organ development. Accumulation of CKX1 in reproductive tissues of transgenic maize (Zea mays) resulted in male-sterile plants. The male development of these plants was restored by applications of kinetin and thidiazuron. Similarly, expression of gai specifically in anthers and pollen of tobacco (Nicotiana tabacum) and Arabidopsis resulted in the abortion of these respective tissues. The gai-induced male-sterile phenotype exhibited by the transgenic plants was reversible by exogenous applications of kinetin. Our results provide molecular evidence of the involvement of cytokinin and GA in male development and support the hypothesis that the male development is controlled in concert by multiple hormones. These studies also suggest a potential method for generating maintainable male sterility in plants by using existing agrochemicals that would reduce the expense of seed production for existing hybrid crops and provide a method to produce hybrid varieties of traditionally non-hybrid crops.

Adenine↗

Upregulation of an Arabidopsis RING-H2 gene, XERICO, confers drought tolerance through increased abscisic acid biosynthesis.

RING (really interesting new gene) zinc-finger proteins have important regulatory roles in the development of a variety of organisms. The XERICO gene encodes a small protein (162 amino acids) with an N-terminal trans-membrane domain and a RING-H2 zinc-finger motif located at the C-terminus. In silico gene-expression analysis indicated that XERICO is induced by salt and osmotic stress. Compared with wild-type (WT) Arabidopsis plants, transgenic plants overexpressing XERICO (35S::XERICO) exhibited hypersensitivity to salt and osmotic stress and exogenous abscisic acid (ABA) during germination and early seedling growth. When subjected to a drought treatment, transcriptional upregulation of a key ABA-biosynthesis gene, AtNCED3, was much faster and stronger in 35S::XERICO plants compared with WT plants. Further, upregulation of XERICO substantially increased cellular ABA levels. The adult 35S::XERICO plants, in contrast to early seedling growth, showed a marked increase in their tolerance to drought stress. Yeast two-hybrid screening indicated that XERICO interacts with an E2 ubiquitin-conjugating enzyme (AtUBC8) and ASK1-interacting F-box protein (AtTLP9), which is involved in the ABA-signaling pathway. Affymetrix GeneChip array analysis showed that the expressions of many of the genes involved in the biosynthesis of plant hormones (e.g. ethylene, brassinosteroid, gibberellic acid) were significantly changed in the 35S::XERICO plants. These results suggest that the homeostasis of various plant hormones might be altered in 35S::XERICO plants, possibly by overaccumulation of ABA.

Abscisic Acid↗

Focus issue: plant communication.

This week's issues of Science and Science's STKE focus on plant signaling with an emphasis on volatile organic compounds. Science emphasizes the chemistry of and information encoded by plant volatiles and how plants, plant pathogens, or humans utilize this aromatic information. Articles and resources at STKE highlight the evolution of olfactory signal transduction, describe the biosynthetic and signaling pathways of the plant hormone jasmonate, and provide an animated overview of how the plant hormone auxin regulates gene expression.

Arabidopsis Proteins↗

Diversity and unity in the nuclear hormone receptors: a terpenoid receptor superfamily.

The remarkable structural unity among the different members of the nuclear hormone receptor superfamily stands in striking contrast to the diversity of the chemical structures of their ligands. Of the three currently known classes of ligands, steroids, retinoids, and thyroid hormones, the first two share a common biosynthetic pathway. Both are terpenes, which are derived by assembly of isoprene units. This biosynthetic link suggests that the receptors for three other classes of terpenoid hormones, the insect juvenile hormones and the plant hormones gibberellic acid and abscissic acid, may also be members of the superfamily. A number of putative nuclear hormone receptors that do not have known ligands have been isolated. At least some of the ligands for these orphan members of the receptor superfamily may be found on the list of biologically active terpenes. Finally, the terpenoid connection raises interesting issues for the evolution of the receptor superfamily.

Animals↗

Small molecule approaches in plants.

Small molecules offer exciting opportunities for plant science. So far, bioactive small molecules have been identified as plant hormones, herbicides, growth regulators, or taken from animal research. Recently, plant scientists have started to explore further the chemical space for novel modulators of plant hormone signalling, and have followed up this work with exciting discoveries illustrating the potential of small molecules such as brassinazole and sirtinol. New chemical genetic screens have been designed to generate chemical tools for the investigation of membrane trafficking, gravitropism and plant immunity. Further novel 'chemetic' tools to identify targets and modes of action are currently generated through an intimate interdisciplinary collaboration between biologists and small molecule chemists.

Benzamides↗

Hybrid hybridoma cell lines which express anti-zeatin riboside, anti-abscisic Acid, and hybrid antibodies.

Hybrid hybridoma cell lines that secreted antibodies which reacted with two distinct plant hormones, abscisic acid (ABA) and zeatin riboside (ZR) were prepared and cloned. These cell lines were derived by polyethylene glycol-mediated fusion of HAT-sensitive anti-ZR hybridoma cells with splenocytes harvested from a BALB/c mouse previously immunized with an ABA-bovine serum albumin conjugate. Chromatographic analyses indicated that these lines expressed two different isotypes, each associated with a specific immunologic reactivity, and that the populations of immunoglobulins secreted by these hybridomas included antibodies directed against each individual hapten as well as hybrid molecules which reacted simultaneously with both. Hybrid hybridomas such as these should provide antibody populations useful for simultaneous isolation of multiple plant hormones from individual plant samples.

Journal Article↗