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Restriction primers as short as 6-mers for PCR amplification of bacterial and plant genomic DNA and plant viral RNA.

Amplification of DNA or RNA sequences using the polymerase chain reaction (PCR) or reverse transcriptase PCR (RT-PCR) requires primers of an appropriate length to be designed. Two hexamer restriction primers, denoted as E101 and H301, which correspond to sequences of EcoRI and HindIII recognition sites, respectively, were selected and used as primers in PCR and RT-PCR. We first applied the restriction primers to the plasmid DNA and bacterial (Pseudomonas) and plant (Cymbidium) genomic DNAs. We observed positive DNA amplifications with the recombinant plasmid DNA and bacterial and plant genomic DNAs. Purified viral RNA was used for template in the RT-PCR with the primers and successful DNA amplification was obtained. These results suggest that the 6-mer restriction primers can be useful for new applications in PCR.

DNA Primers↗

[90Sr and 137Cs in higher aquatic plants of the Chernobyl nuclear plant exlusion zone]].

The content of radionuclides 90Sr and 137Cs in higher aquatic plants of water objects within Chernobyl NPP exclusion zone has been analysed. Biodiversity of phytocenose was studied and species-indicators of radioactive contamination were revealed. The seasonal dynamics of radionuclide content in macrophytes was studied and the role of main aquatic plant clumps in processes of 137Cs and 90Sr distribution in abiotic component of biohydrocenose was demonstrated.

Cesium Radioisotopes↗

How plants see themselves--self-incompatibility in flowering plants.

Plants have been propagating themselves by cloning for millennia. It is, however, widely recognised that mixing genes with other individuals of the same species makes better evolutionary sense, as it provides the variation that is the raw material for natural selection. How, then, do some plants prevent self-fertilisation?

Evolution, Molecular↗

Hydrotechnical facilities within the Chernobyl nuclear power plant exclusion zone: impacts on hydrologic regime and plant growth patterns of floodplain water bodies of the Pripyat River.

As result of the Chernobyl nuclear power plant accident the territory of the left-bank flood-lands of the Pripyat River have undergone intensive radionuclide contamination. With the purpose of preventing the washing away of radioactive substances, a complex of flood protection dams was constructed. This construction changed the hydrological regime of these territories and caused overgrowth by higher aquatic plants. Absence of a flowing mode of reservoirs, the stagnant phenomena during spring and seasonal high waters on the embank site have caused amplification of eutrophication processes, swamping and, connected with it, increase of water-marsh floristic complex in the structure of the vegetative cover.

Ecosystem↗

[Interaction between proteinases secreted by the fungal plant pathogen Rhizoctonia solani and natural proteinase inhibitors produced by plants].

The fungal plant pathogen Rhizoctonia solani Kuhn. grown in a medium containing thermostable potato tuber proteins produced proteinases active at moderately alkaline pH values. Electrophoretic analysis in polyacrylamide gel with SDS and copolymerized gelatin showed that the extracellular proteinase complex contained four components that differed in molecular weight. Studies on the action of the exoenzymes on various synthetic substrates indicated that the culture liquid of R. solani contained mainly trypsin-like proteinases. The exoproteinase activity was virtually completely suppressed by trypsin inhibitor proteins isolated from potato tubers and seeds of various legume species. The results suggest that the extracellular proteinases produced by R. solani play a significant role in attacking plant tissue, and natural inhibitors contribute to the protection of Solanaceae and Leguminosae from this fungal pathogen.

Basidiomycota↗

Mining plant diversity: Gerbera as a model system for plant developmental and biosynthetic research.

Gerbera hybrida is a member of the large sunflower family (Asteraceae). Typical of Asteraceae, Gerbera bears different types of flowers in its inflorescence. The showy marginal flowers comprise elongate, ligulate corollas that are female, whereas the central and inconspicuous disc flowers are complete, with both male and female organs. As such, Gerbera offers great potential for comparative developmental research within a single genotype. Moreover, different Gerbera varieties show an impressive spectrum of color patterns, directly displaying responses to developmental cues at all important morphological levels (flower type, flower organ and within organs). Further, Gerbera harbors an arsenal of Asteraceae-type secondary metabolites, not present in other model plants. With powerful reverse genetics methods, a large collection of EST sequences and a new cDNA microarray, Gerbera has become a model plant of the sunflower family.

Asteraceae↗

Lipoxygenase inhibitors from natural plant sources. Part 2: medicinal plants with inhibitory activity on arachidonate 12-lipoxygenase, 15-lipoxygenase and leukotriene receptor antagonists.

The metabolism of arachidonic acid can be catalysed by either one of two enzyme families: the cyclooxygenases or the lipoxygenases. The lipoxygenase enzymes are classed into several subcategories including 5-, 12- and 15-lipoxygenases. The 5-lipoxygenase pathway has been the major focus of study due to the pronounced pro-inflammatory role of leukotrienes and the approval of 5-lipoxygenase inhibitors and leukotriene receptor antagonists for the clinical treatment of asthma. Although less well characterized, the 12-lipoxygenase as well as the 15-lipoxygenase pathway may also play an important role in the progression of human diseases such as cancer, psoriasis and atherosclerosis. The present review article summarizes the findings from an extensive literature search on plants that have been assessed for 12- and 15-lipoxygenase inhibitory activity as well as for leukotriene receptor antagonistic properties. The results are presented in a tabular format, and a discussion about promising plant species and natural compounds as well as relevant in vitro assays are included in this article.

Anti-Asthmatic Agents↗

S-Adenosyl-L-methionine:salicylic acid carboxyl methyltransferase, an enzyme involved in floral scent production and plant defense, represents a new class of plant methyltransferases.

S-Adenosyl-L-methionine:salicylic acid carboxyl methyltransferase (SAMT) was partially purified from petals of the annual California plant Clarkia breweri. SAMT catalyzes the formation of methylsalicylate, an important floral scent compound in C. breweri, from salicylic acid and S-adenosyl-L-methionine (SAM). The native enzyme is a dimer with a subunit molecular weight of 40.3 kDa, and it has a Km for salicylic acid of 24 microM and a Km for SAM of 9 microM. A cDNA encoding SAMT was isolated from a C. breweri cDNA library prepared from floral mRNA. The sequence of the protein encoded by SAMT cDNA shows no significant sequence similarity to any protein in the data bank whose biochemical function is known. It does show significant sequence similarity (20-40% identity) to proteins encoded by at least seven Arabidopsis thaliana genes whose sequences have recently been determined in large-scale sequencing projects. The C. breweri SAMT cDNA was expressed in E. coli and the bacterial cells synthesized a functional SAMT protein with properties nearly identical to those of the plant-purified enzyme.

Amino Acid Sequence↗

Short-term tests of estrogenic potential of plant stanols and plant stanol esters.

To test for potential estrogenic activity of plant stanols and plant stanol esters, two short-term tests were performed. These were the E-screen test, which measures a substance's ability to induce proliferation of estrogen-responsive human breast adenocarcinoma (MCF-7) cells in culture, and an in vivo test, which measures uterotrophic activity in immature female rats fed the test substance. Four samples of vegetable oil-derived stanols (containing 88-99% stanols) were tested in the E-screen test, and one sample of wood-derived and one of vegetable oil-derived stanol fatty acid esters were tested in the in vivo test. In the E-screen test, the positive control substance, 17beta-estradiol, at 100 pM, produced a statistically significant, 11.6-fold increase in cell proliferation, as measured by sulforhodamine B staining. None of the stanol preparations produced any increase in cell proliferation when tested at 1, 10, and 100 microM. The highest dose of each stanol sample was associated with microscopic evidence of cytotoxicity and crystalline precipitation in the culture dishes. In the in vivo test, the positive control compound, diethylstilbestrol, produced a significant, dose-related increase in absolute and relative uterus weight in young female rats (17 days old at the start of treatment) fed the compound at 5, 10, and 20 ppb in the diet for 4 days. Neither of the two stanol ester preparations caused any significant change in absolute or relative uterus weight when fed at a concentration of 8.3% in the diet for 4 days. Thus, under the conditions of testing used, neither the free stanols nor the stanol fatty acid ester preparations showed evidence of estrogenic or uterotrophic activity.

Analysis of Variance↗

The sensitizing capacity of Compositae plants. VI. Guinea pig sensitization experiments with ornamental plants and weeds using different methods.

Experimental studies in guinea pigs using ether extracts of 20 different species of the Compositae plant family were carried out with the open epicutaneous method (OET) and the guinea pig maximization test (GPMT). The results demonstrate that Cnicus benedictus (blessed thistle), Chrysanthemum leucanthemum (marguerite, ox-eye daisy) and Helianthus debilis (dwarf sunflower) are strong sensitizers while Helenium amarum (bitterweed), Gaillardia amblyodon (blanket flower), Artemisia ludoviciana (prairie sage), Ambrosia trifida (giant ragweed) and Solidago virgaurea (goldenrod) are medium sensitizers. Twelfe species revealed only a weak or no sensitizing capacity; among those were corn flower, wormwood, mugwort, coltsfoot and dandelion. Cross-reactivities were observed in a considerable number of the investigated plant species. The sensitizing power as well as the observed cross-reactions depend on the occurrence of sesquiterpene lactones which have an alpha-methylene group exocyclic to the lactone in common ("immunologic requisite"). As a practical consequence, patients suffering from allergic contact dermatitis due to Compositae species are strictly requested to avoid contact with the offending species and all related species to prevent recurrences of their skin lesions.

Allergens↗

Powerful hepatoprotective and hepatotoxic plant oligostilbenes, isolated from the Oriental medicinal plant Vitis coignetiae (Vitaceae).

The methanol extract of the Oriental medicinal plant Vitis coignetiae (Vitaceae) showed hepatoprotective activity in the in vitro assay method using primary cultured rat hepatocytes. Activity-guided fractionation of the extract afforded epsilon-viniferin as an active principle. The protective effect of epsilon-viniferin against mice carbon tetrachloride-induced hepatic injury in mice was shown by serum enzyme assay as well as by pathological examination. In addition to epsilon-viniferin, plant oligostilbenes, ampelopsins A, C, F and the mixture of vitisin A and cis-vitisin A were also present in the extract. Among them, ampelopsin C and the mixture of vitisin A and cis-vitisin A were found to be powerful hepatotoxins.

Animals↗

Plant regeneration from callus of apomictic and sexual lines of Paspalum simplex and RFLP analysis of regenerated plants.

Culture conditions have been established for the induction of callus from different explants of Paspalum simplex. Fast-growing calli were obtained from hypocotyls and roots excised from 5-day-old seedlings on culture medium containing 2,4-dichlorophenoxyacetic acid and kinetin. Rapid plant regeneration from both apomictic and sexual lines was achieved when the medium was supplemented with alpha-naphthaleneacetic acid and benzylaminopurine. Restriction fragment length polymorphism analysis of the apomixis-controlling region of the regenerated plants showed an absence of restriction site variation for the loci analysed, whereas various degrees of variation were detected for the DNA methylation sites of the same loci.

Adenine↗

A new tool for plant cell biology: in vivo antibody uptake in plant protoplasts.

We report on the in vivo uptake of antibodies into plant protoplasts. When protoplasts of sunflower, Arabidopsis or tobacco were incubated in vivo with an antibody, this antibody was detected by immunofluorescence in the cytoplasm and/or the nucleus, depending on the location of the target protein. Furthermore, when protoplasts were cultured in the presence of antibodies, specific effects were observed. Incubation with antibodies raised against p34cdc2 led to a strong inhibition of the division rate, and a decrease in the average DNA content of protoplasts. With antibodies against HaWLIM1, a LIM domain protein of the CRP type, a negative effect on actin organisation was observed. We conclude that antibodies can penetrate plant protoplasts in vivo, and thus may be used as powerful tools for the study of protein function.

Animals↗

Achieving desired plant growth regulator levels in liquid plant tissue culture media that include activated carbon.

This paper is part of a series considering the impact of activated carbon (AC) on the composition of plant tissue culture media. Using liquid culture media for initiation of Norway spruce embryogenic tissue and eight different ACs, we present a method for achieving target plant growth regulator (PGR) levels in AC-containing medium based on sorption isotherms for individual PGRs. Linear relationships were found between PGR adsorption and specific BET (Brunauer, Emmett, Teller theory) surface area and specific total pore volume of AC. When using a new AC, this linear relationship allows one to achieve multiple PGR levels similar to historic levels through adjustment of the mass of AC based on its relative BET surface area or relative total pore volume. Target levels of PGRs and an initiation success similar to that in medium without AC were achieved with several different AC types when AC mass was adjusted on the basis of pore volume.

Cell Culture Techniques↗

Chemically mediated host-plant selection by the milfoil weevil: a freshwater insect-plant interaction.

The milfoil weevil Euhrychiopsis lecontei is a specialist aquatic herbivore that feeds, oviposits, and mates on the invasive freshwater macrophyte Myriophyllum spicatum. We characterized the weevil's preference for M. spicatum, and through bioassay-driven fractionation, isolated and identified two chemicals released by M. spicatum that attract E. lecontei. Mass spectrometry and nuclear magnetic resonance spectroscopy were used to identify the attractive compounds as glycerol and uracil. Dose-response curves for glycerol and uracil indicated that weevil preference increased as sample concentration increased. Weevils were attracted to a crude sample of M. spicatum-released chemicals from 0.17 to 17 mg/l, to glycerol from 18 to 1800 microM (0.0017-0.17 mg/l), and to uracil from 0.015 to 15 microM (0.00014-1.4 mg/l). Although glycerol and uracil are ubiquitous, weevils are likely responding to high concentrations that are released as a result of the rapid growth of M. spicatum. Uracil concentration was greater in the exudates of M. spicatum than other Myriophyllum spp. E. lecontei was attracted to glycerol at a concentration similar to that at which terrestrial insects are attracted to sugar alcohols. This is the first example of a freshwater specialist insect being attracted to chemicals released by its host plant. Analysis of the water milfoil-weevil interaction provides further understanding as to how insects locate their host plants in aquatic systems.

Chromatography, DEAE-Cellulose↗

Plant cytoskeleton: reinforcing lines of division in plant cells.

Cytokinesis in plants has unique features concerned with defining and maintaining the line of cell division. Recent studies have identified key cytoskeletal components and events that help to ensure the fidelity of cytokinesis in higher plants.

Cell Cycle Proteins↗

Soil-to-plant transfer of radiocaesium for selected tropical plant species in Bangladesh.

Soil-to-plant transfer factors (TF) of radiocaesium (137Cs) were determined under field condition for grassy vegetation grown in Bangladesh at contaminated land in the Atomic Energy Research Establishment (AERE) campus. TF values for rice, grass and grassy/root vegetations grown in the same type of soil were also measured under pot condition. TF values of 137Cs for grassy vegetation (2.4 x 10(-2) -4.2 x 10(-2) with an average of 3.1 x 10(-2) +/-0.005) obtained under field condition were slightly lower than the values for grass and grassy/root vegetations (2.9 x 10(-2) -6.6 x 10(-2) with an average of 4.8 x 10(-2) +/-0.01 for grass and grassy vegetations and 2.3 x 10(-2) -5.6 x 10(-2) with an average of 4.0 x 10(-2) +/-0.009 for root vegetations, respectively) obtained under pot condition. However, TF values (9.0 x 10(-3) -2.6 x 10(-2) with an average of 1.9 x 10(-2) +/-0.004) obtained for rice were about a factor of 4 lower than the values obtained for grass and grassy/root vegetations. When the properties of the AERE soils as input parameters were used in the soil-plant transfer model of Absalom, the estimated TF values (4.5 x 10(-2) -6.7 x 10(-2) with an average of 5.3 x 10(-2) +/-0.006) were consistent with the measured values obtained for grass and grassy vegetations under pot condition, however, the model overestimates the TF values for rice.

Aluminum Silicates↗

Identification and characterization of plant agmatine iminohydrolase, the last missing link in polyamine biosynthesis of plants.

The cloning, expression and characterization of plant agmatine iminohydrolase (AIH, also known as agmatine deiminase, EC 3.5.3.12) is described. Recombinant AIH of Arabidopsis thaliana forms dimers and catalyzes the specific conversion of agmatine to N-carbamoylputrescine and ammonia. Biochemical data suggested that cysteine side chains are involved in catalysis. However, site-directed mutagenesis of the two highly conserved cysteine residues of AIH showed that these cysteines are important but not essential for activity, arguing against a thioester substrate-enzyme intermediate during catalysis. This work represents the completion of the cloning of the arginine decarboxylase pathway genes of higher plants.

Ammonia↗