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PubMed1999

[Deletion analysis and functional studies of the promoter from commelina yellow mottle virus].

Commelina Yellow Mottle Virus(CoYMV) is a double-stranded, circular DNA virus and its promoter could direct GUS gene specifically expressing in phloem tissue of transgenic tobacco plants. To determine the optimal promoter sequence for pholem-specific gene expression, CoYMV promoter was deleted from its 5 end to form promoter fragments with 5 different lengths. Chimeric GUS genes were constructed using the promoter deletion based on the binary vector pBI121. Transgenic tobacco plants evidenced by PCR analysis were obtained with each kind of chimeric GUS gene structure by Agrobacterium mediated transformation. The results of GUS activity assay and histo-chemical staining showed that most of the chimeric GUS genes were expressed in transgenic plants. The GUS activity with the promoter deleted to -870 bp was about 78% higher than that of the full length promoter(1040 bp) and was a little higher than that of the promoter deleted to -585 bp, but the difference is not significant. The GUS activity reduced significantly when the promoter was deleted to -447 bp or -232 bp, whereas the property of phloem-specific expression pattern was still retained. When the promoter was deleted to -44 bp, just upstream adjacent to the TATA box, its tissue-specificity was lost and the activity was reduced to undetectable level. These results suggest that the region between -870 bp-232 bp and downstream of -232 bp of CoYMV promoter could be responsible for promoter activity and tissue specific expression, respectively. A negative regulation sequence might exist upstream of -870 bp of the CoYMV promoter. Therefore, we recommend that the optional CoYMV promoter sequence for phloem specific expression could be downstream from -870 bp or -585 bp. In comparison with CaMV 35S promoter, the GUS activity when driven by -870 bp CoYMV promoter was about 70% of that when driven by the 35S promoter. Considering the fact that 35S promoter-GUS gene is constitutively expressed, while the CoYMV promoter-GUS gene is expressed only in phloem tissues, the activity of the latter in phloem may be the same with or even higher than that of the 35S promoter.

Badnavirus↗
PubMed1994

[Active constituents of Commelina communis L].

According to the pharmacological results five compounds were isolated from the herb of Commelina communis. Based on physico-chemical constants and spectral data, four of them were identified as n-triacontanol, p-hydroxycinnamic acid, daucosteril and D-mannitol. p-hydroxycinnamic acid shows antibacterial activity and D-mannitol shows antitussive effect.

Animals↗
PubMed1988

Enhancement of the Stomatal Response to Blue Light by Red Light, Reduced Intercellular Concentrations of CO(2), and Low Vapor Pressure Differences.

The effects of environmental parameters on the blue light response of stomata were studied by quantifying transient increases in stomatal conductance in Commelina communis following 15 seconds by 0.100 millimole per square meter per second pulses of blue light. Because conductance increases were not observed following red light pulses of the same or greater (30 seconds by 0.200 millimole per square meter per second) fluences, the responses observed could be reliably attributed to the specific blue light response of the guard cells, rather than to guard cell chlorophyll. In both Paphiopedilum harrisianum, which lacks guard cell chloroplasts, and Commelina, the blue light response was enhanced by 0.263 millimole per square meter per second continuous background red light. Thus, the blue light response and its enhancement do not require energy derived from red-light-driven photophosphorylation by the guard cell chloroplasts. In Commelina, reduction of the intercellular concentration of CO(2) by manipulation of ambient CO(2) concentrations resulted in an enhanced blue light response. In both Commelina and Paphiopedilum, the blue light response was decreased by an increased vapor pressure difference. The magnitude of blue-light-specific stomatal opening thus appears to be sensitive to environmental conditions that affect the carbon and water status of the plant.

Journal Article↗
PubMed2004

Responses of antioxidative enzymes to accumulation of copper in a copper hyperaccumulator of Commoelina communis.

Discovery of a copper hyperaccumulator is very important for phytoremediation of copper-contaminated soil. In the present study a link was established between the copper accumulation in hyperaccumulator and that in nonaccumulator species of Commelina communis and its responses of antioxidative enzymes, including superoxide dismutase, guaiacol peroxidase, and ascorbate peroxidase. It was verified that copper exerted little physiological damage to copper hyperaccumulator species of Commelina communis at copper accumulation of >1,000 microg/g in dry leaf tissue. However, in nonaccumulator species of Commelina communis superoxide dismutase, guaiacol peroxidase, and ascorbate peroxidase were activated, and malondialdehyde content was increased, which were symptoms of physiological damage by copper intoxication. Therefore, antioxidative enzymes can be used as an indicator of copper toxicity before the visible symptoms can be observed.

Ascorbate Peroxidases↗
PubMed2000

Macrophomate synthase: characterization, sequence, and expression in Escherichia coli of the novel enzyme catalyzing unusual multistep transformation of 2-pyrones to benzoates.

Macrophoma commelinae isolated from spots on leaves of Commelina communis has the ability to transform 5-acetyl-4-methoxy-6-methyl-2-pyrone (1) to 4-acetyl-3-methoxy-5-methylbenzoic acid (macrophomic acid, 2). This biotransformation includes the condensation of the 2-pyrone ring with a C3-unit precursor to form a substituted benzoic acid. We optimized conditions for induction of enzyme activity in M. commelinae, identified oxalacetate as a C3-unit precursor with cell extract, and purified the novel enzyme, macrophomate synthase. Oxalacetate inhibited the enzyme activity at a concentration higher than 5 mM, and magnesium chloride stimulated the enzyme activity. Kinetic analyses gave K(m) of 1.7 mM for 1 at 5 mM oxalacetate, K(m) of 1.2 mM for oxalacetate at 5 mM 1, and k(cat) of 0.46 s(-1) per subunit. Pyruvate was a weak substrate, with K(m) of 35.2 mM and k(cat) of 0.027 s(-1) at 5 mM 1. We cloned and sequenced a cDNA encoding the macrophomate synthase. The cDNA of 1,225 bp contained an open reading frame that encoded a polypeptide of 339 amino acid residues and 36,244 Da, the sequence of which showed no significant similarity with known proteins in a homology search with BLAST programs. Transformed E. coli cells carrying the cDNA encoding the mature protein of macrophomate synthase overproduced macrophomate synthase under the control of the T7 phage promoter induced by IPTG. The purified enzyme showed the same values of K(m) and optimum pH as the native macrophomate synthase.

Amino Acid Sequence↗
PubMed1999

Characterization of stomatal closure caused by ultraviolet-B radiation

The effects of ultraviolet-B (UV-B) radiation on stomatal conductance (g(s)) in pea (Pisum sativum L.), commelina (Commelina communis L.), and oilseed rape (Brassica napus L.) plants were investigated. Plants were grown in a greenhouse either with three different high ratios of UV-B to photosynthetically active radiation or with no UV-B radiation. Pea plants grown in the highest UV-B radiation (0.63 W m(-2)) exhibited a substantial decrease of adaxial and abaxial g(s) (approximately 80% and 40%, respectively). With growth in 0.30 W m(-2) of UV-B adaxial g(s) was decreased by 23%, with no effect on abaxial g(s), and lower UV-B irradiance of 0.21 W m(-2) had no effect on either surface. Although abaxial g(s) increased when leaves were turned over in control plants, it did not in plants grown with the highest UV-B. Adaxial g(s) in commelina and oilseed rape also decreased on exposure to high UV-B (0.63 W m(-2)). For previously unexposed pea plants the time course of the effect of UV-B on g(s) was slow, with a lag of approximately 4 h, and a time constant of approximately 3 h. We conclude that there is a direct effect of UV-B on stomata in addition to that caused by changes in mesophyll photosynthesis.

Journal Article↗
PubMed2006

In vitro evaluation of effects of two Ghanaian plants relevant to wound healing.

Commelina diffusa and Spathodea campanulata are used as wound-healing agents in Ashanti traditional medicine in Ghana. The methanol extracts of Commelina diffusa herb and Spathodea campanulata bark showed some level of antimicrobial activity with C. diffusa exhibiting selective antifungal activity against Trichophyton species. The extracts reduced the peroxidation of bovine brain extract with an IC(50) value of 1.39 mg/mL and 0.24 mg/mL, respectively. In addition the extracts also exhibited significant antioxidant activity by protecting MRC-5 cells from hydrogen peroxide induced oxidant injury at concentrations between 1 microg/mL and 10 microg/mL. The extracts showed no inhibition of NF-kappaB at 100 microg/mL. The antioxidant activities and antimicrobial activities suggest that the use of the plants in wound healing may be based on antioxidant and antiseptic effects of its constituents.

Animals↗
PubMed1994

Inhibition of inward K+ channels and stomatal response by abscisic acid: an intracellular locus of phytohormone action.

Abscisic acid (ABA), a plant hormone whose production is stimulated by water stress, reduces the apertures of stomatal pores in the leaf surface, thereby lessening transpirational water loss. It has been thought that inhibition of stomatal opening and promotion of stomatal closure by ABA are initiated by the binding of extracellular ABA to a receptor located in the guard-cell plasma membrane. However, in the present research, we employ three distinct experimental approaches to demonstrate that ABA can act from within guard cells to regulate stomatal apertures. (i) The extent to which ABA inhibits stomatal opening and promotes stomatal closure in Commelina communis L. is proportional to the extent of ABA uptake, as assayed with [3H]ABA. (ii) Direct microinjection of ABA into the cytoplasm of Commelina guard cells precipitates stomatal closure. (iii) Application of ABA to the cytosol of Vicia faba L. guard-cell protoplasts via patch-clamp techniques inhibits inward K+ currents, an effect sufficient to inhibit stomatal opening. These results demonstrate an intracellular locus of phytohormone action and imply that the search for hormone receptor proteins should be extended to include intracellular compartments.

Abscisic Acid↗
PubMed1995

Anion-Channel Blockers Inhibit S-Type Anion Channels and Abscisic Acid Responses in Guard Cells.

The effects of anion-channel blockers on light-mediated stomatal opening, on the potassium dependence of stomatal opening, on stomatal responses to abscisic acid (ABA), and on current through slow anion channels in the plasma membrane of guard cells were investigated. The anion-channel blockers anthracene-9-carboxylic acid (9-AC) and niflumic acid blocked current through slow anion channels of Vicia faba L. guard cells. Both 9-AC and niflumic acid reversed ABA inhibition of stomatal opening in V. faba L. and Commelina communis L. The anion-channel blocker probenecid also abolished ABA inhibition of stomatal opening in both species. Additional tests of 9-AC effects on stomatal aperture in Commelina revealed that application of this anion-channel blocker allowed wide stomatal opening under low (1 mM) KCI conditions and increased the rate of stomatal opening under both low and high (100 mM) KCI conditions. These results indicate that anion channels can function as a negative regulator of stomatal opening, presumably by allowing anion efflux and depolarization, which prohibits ion up-take in guard cells. Furthermore, 9-AC prevented ABA induction of stomatal closure. A model in which ABA activation of anion channels contributes a rate-limiting mechanism during ABA-induced stomatal closure and inhibition of stomatal opening is discussed.

Journal Article↗
PubMed1981

Electrical potentials in stomatal complexes.

Guard cells of several species, but predominantly Commelina communis, were impaled by micropipette electrodes and potential differences measured that occurred between cell compartments and the flowing bathing medium. The wall developed a Donnan potential that was between -60 and -70 millivolt in 30 millimolar KCl at pH 7. The density of the fixed charges ranged from 0.3 to 0.5 molar; its dependence on pH was almost identical with the titration curve of authentic polygalacturonic acid. The vacuolar potential of guard cells of Commelina communis L., Zea mays L., Nicotiana glauca Graham, Allium cepa L., and Vicia faba L. was between -40 and -50 millivolt in 30 millimolar KCl when stomata were open and about -30 millivolt when stomata were closed. The vacuolar potential of guard cells of C. communis was almost linearly related to stomatal aperture and responded to changes in the ionic strength in the bathing medium in a Nernstian manner. No specificity for any alkali ion (except Li(+)), ammonium, or choline appeared. Lithium caused hyperpolarization. Calcium in concentrations between 1 and 100 millimolar in the medium led to stomatal closure, also caused hyperpolarization, and triggered transient oscillations in the intracellular potential. Gradients in the electrical potential existed across stomatal complexes with open pores. When stomata closed, these gradients almost disappeared or slightly reverted; all epidermal cells were then at potentials near -30 millivolt in 30 millimolar KCl.

Journal Article↗
PubMed2002

Responses of photosynthetic electron transport in stomatal guard cells and mesophyll cells in intact leaves to light, CO2, and humidity.

High-resolution images of the chlorophyll fluorescence parameter Fq'/Fm' from attached leaves of commelina (Commelina communis) and tradescantia (Tradescantia albiflora) were used to compare the responses of photosynthetic electron transport in stomatal guard cell chloroplasts and underlying mesophyll cells to key environmental variables. Fq'/Fm' estimates the quantum efficiency of photosystem II photochemistry and provides a relative measure of the quantum efficiency of non-cyclic photosynthetic electron transport. Over a range of light intensities, values of Fq'/Fm' were 20% to 30% lower in guard cell chloroplasts than in mesophyll cells, and there was a close linear relationship between the values for the two cell types. The responses of Fq'/Fm' of guard and mesophyll cells to changes of CO2 and O2 concentration were very similar. There were similar reductions of Fq'/Fm' of guard and mesophyll cells over a wide range of CO2 concentrations when the ambient oxygen concentration was decreased from 21% to 2%, suggesting that both cell types have similar proportions of photosynthetic electron transport used by Rubisco activity. When stomata closed after a pulse of dry air, Fq'/Fm' of both guard cell and mesophyll showed the same response; with a marked decline when ambient CO2 was low, but no change when ambient CO2 was high. This indicates that photosynthetic electron transport in guard cell chloroplasts responds to internal, not ambient, CO2 concentration.

Algorithms↗
PubMed2004

A conserved functional role of pectic polymers in stomatal guard cells from a range of plant species.

Guard cell walls combine exceptional strength and flexibility in order to accommodate the turgor pressure-driven changes in size and shape that underlie the opening and closing of stomatal pores. To investigate the molecular basis of these exceptional qualities, we have used a combination of compositional and functional analyses in three different plant species. We show that comparisons of FTIR spectra from stomatal guard cells and those of other epidermal cells indicate a number of clear differences in cell-wall composition. The most obvious characteristics are that stomatal guard cells are enriched in phenolic esters of pectins. This enrichment is apparent in guard cells from Vicia faba (possessing a type I cell wall) and Commelina communis and Zea mays (having a type II wall). We further show that these common defining elements of guard cell walls have conserved functional roles. As previously reported in C. communis, we show that enzymatic modification of the pectin network in guard cell walls in both V. faba and Z. mays has profound effects on stomatal function. In all three species, incubation of epidermal strips with a combination of pectin methyl esterase and endopolygalacturonase (EPG) caused an increase in stomatal aperture on opening. This effect was not seen when strips were incubated with EPG alone indicating that the methyl-esterified fraction of homogalacturonan is key to this effect. In contrast, arabinanase treatment, and incubation with feruloyl esterase both impeded stomatal opening. It therefore appears that pectins and phenolic esters have a conserved functional role in guard cell walls even in grass species with type II walls, which characteristically are composed of low levels of pectins.

Commelina↗
PubMed2004

Characterization and activity enhancement of the phloem-specific pumpkin PP2 gene promoter.

The promoter of the pumpkin (Cucurbita moschata) PP2 gene (designated NP) was isolated from the restriction enzyme-digested genomic DNA pool by genome walking and its activity and phloem specificity were examined in transgenic tobacco plants by using GUS as a reporter. Deletion analysis of the promoter revealed that the 473-bp fragment (-465 to + 8 relative to the transcription start site; designated as NPII) exhibited similar activity as the full-length NP promoter and retained its phloem specificity. Furthermore, the sequence from -465 to -171 was shown to contain positive regulatory cis-elements for the promoter activity. An enhanced NP promoter was constructed by duplicating the sequence -465 to -85, and its activity in phloem tissue was shown to be higher than that of the Commelina Yellow Mottle Virus (CoYMV) promoter or a chimeric promoter consisting of the double enhancer sequence from the Cauliflower Mosaic Virus (CaMV) 35S promoter fused upstream to the NPII fragment.

Agrobacterium tumefaciens↗
PubMed2005

Isolation and stomatal opening activity of two oxylipins from Ipomoea tricolor.

Bioassay-guided fractionation of a 50% MeOH extract of Ipomoea tricolor enabled the isolation of two oxylipins, cis-12-oxophytodienoic acid (OPDA, 1) and a novel monogalactosylmonoacylglyceride (2) containing OPDA, that acted as inducers of stomatal opening. These oxylipins enhanced stomatal opening of Commelina communis in darkness at micromolar concentrations.

Commelina↗
PubMed2005

Identification of weed plants excluding the uptake of heavy metals.

Using the field pot-culture and sample-analysis method, 54 weed species belonging to 20 families and 31 weed species belonging to 17 families were systematically examined as to whether they can exclude the uptake of heavy metals. After a systematic identification, it was determined that Oenothera biennis and Commelina communis were Cd-excluders and Taraxacum mongolicum was a Zn-excluder. O. biennis is a potential Cd-excluder, but also a potential Cu-excluder. The research raises the possibility of making a major breakthrough in the application of metal excluders for safe agro-production in the future.

Cadmium↗