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PubMedDate not supplied

Hybridization and backcrossing between transgenic oilseed rape and two related weed species under field conditions.

Determining the frequency of crop-wild transgene flow under field conditions is a necessity for the development of regulatory strategies to manage transgenic hybrids. Gene flow of green fluorescent protein (GFP) and Bacillus thuringiensis (Bt) transgenes was quantified in three field experiments using eleven independent transformed Brassica napus L. lines and the wild relatives, B. rapa L. and Raphanus raphanistrum L. Under a high crop to wild relative ratio (600:1), hybridization frequency with B. rapa differed among the individual transformed B. napus lines (ranging from ca. 4% to 22%), however, this difference could be caused by the insertion events or other factors, e.g., differences in the hybridization frequencies among the B. rapa plants. The average hybridization frequency over all transformed lines was close to 10%. No hybridization with R. raphanistrum was detected. Under a lower crop to wild relative ratio (180:1), hybridization frequency with B. rapa was consistent among the transformed B. napus lines at ca. 2%. Interspecific hybridization was higher when B. rapa occurred within the B. napus plot (ca. 37.2%) compared with plot margins (ca. 5.2%). No significant differences were detected among marginal plants grown at 1, 2, and 3 m from the field plot. Transgene backcrossing frequency between B. rapa and transgenic hybrids was determined in two field experiments in which the wild relative to transgenic hybrid ratio was 5-15 plants of B. rapa to 1 transgenic hybrid. As expected, ca. 50% of the seeds produced were transgenic backcrosses when the transgenic hybrid plants served as the maternal parent. When B. rapa plants served as the maternal parent, transgene backcrossing frequencies were 0.088% and 0.060%. Results show that transgene flow from many independent transformed lines of B. napus to B. rapa can occur under a range of field conditions, and that transgenic hybrids have a high potential to produce transgenic seeds in backcrosses.

Agriculture↗

PubMedDate not supplied

Lack of stable inheritance of introgressed transgene from oilseed rape in wild radish.

Hybridization of Brassica napus L. (oilseed rape) and Raphanus raphanistrum L. (wild radish) has been demonstrated, and may be the first step towards introgression of transgenes in this wild relative. If wild radish were to display a new adaptive advantage by expressing the transgene, this could modify the ecological balance of species within the agro-ecosystem. To determine if transgenes remained stable in the hybrid, the frequency of herbicide resistance was studied over four advanced generations of hybrid progeny (G8 to G11) that were subjected to herbicide selection pressure. It is expected that hemizygous resistant plants containing an herbicide resistance transgene back-crossed to wild radish would have 50% resistant progeny. In each of the G8 to G11 generations, only 18% of the progeny from resistant plants were resistant. The chromosome complement of herbicide-susceptible progenies, analyzed at G9, was not different from that of wild populations of wild radish. Herbicide-resistant G9 progeny showed higher chromosome instability, and one third of the progeny contained a supernumerary chromosome. These results suggest that in the presence of herbicide selection pressure, the transgene for herbicide resistance would be maintained despite a lack of stabilized introgression. In the absence of selection, the frequency of resistance in the population is expected to decline.

Agriculture↗

PubMed2003

Inhibition of mutagenicity of food-derived heterocyclic amines by sulphoraphene--an isothiocyanate isolated from radish.

The naturally derived isothiocyanate, sulphoraphene [4-isothiocyanato-(1R)-(methylsulphinyl)-1-(E)-butene], isolated from seeds of radish ( Raphanus sativus L., Cruciferae) was investigated for its antigenotoxic effects against a battery of cooked food mutagens (heterocyclic amines) in the Ames Salmonella/reversion assay using Salmonella typhimurium TA98 (frame-shift mutation sensitive) and TA100 (base -pair mutation sensitive) bacterial strains in the presence of Aroclor 1254 induced rat liver S9. Results of the present in vitro anti-mutagenicity studies using the base-pair mutation sensitive strain TA100, strongly suggest that sulphoraphene is a potent inhibitor of the S9-mediated mutagenicity of all the tested heterocyclic amines (60 - 75 % inhibition at a dose of 500 nmol/plate).

Amines↗

PubMed2004

Bayesian analysis of botanical epidemics using stochastic compartmental models.

A stochastic model for an epidemic, incorporating susceptible, latent, and infectious states, is developed. The model represents primary and secondary infection rates and a time-varying host susceptibility with applications to a wide range of epidemiological systems. A Markov chain Monte Carlo algorithm is presented that allows the model to be fitted to experimental observations within a Bayesian framework. The approach allows the uncertainty in unobserved aspects of the process to be represented in the parameter posterior densities. The methods are applied to experimental observations of damping-off of radish (Raphanus sativus) caused by the fungal pathogen Rhizoctonia solani, in the presence and absence of the antagonistic fungus Trichoderma viride, a biological control agent that has previously been shown to affect the rate of primary infection by using a maximum-likelihood estimate for a simpler model with no allowance for a latent period. Using the Bayesian analysis, we are able to estimate the latent period from population data, even when there is uncertainty in discriminating infectious from latently infected individuals in data collection. We also show that the inference that T. viride can control primary, but not secondary, infection is robust to inclusion of the latent period in the model, although the absolute values of the parameters change. Some refinements and potential difficulties with the Bayesian approach in this context, when prior information on parameters is lacking, are discussed along with broader applications of the methods to a wide range of epidemiological systems.

Algorithms↗

PubMed2006

Population size and relatedness affect fitness of a self-incompatible invasive plant.

One of the lingering paradoxes in invasion biology is how founder populations of an introduced species are able to overcome the limitations of small size and, in a "reversal of fortune," proliferate in a new habitat. The transition from colonist to invader is especially enigmatic for self-incompatible species, which must find a mate to reproduce. In small populations, the inability to find a mate can result in the Allee effect, a positive relationship between individual fitness and population size or density. Theoretically, the Allee effect should be common in founder populations of self-incompatible colonizing species and may account for the high rate of failed introductions, but little supporting evidence exists. We created a field experiment to test whether the Allee effect affects the maternal fitness of a self-incompatible invasive species, wild radish (Raphanus sativus). We created populations of varying size and relatedness. We measured maternal fitness in terms of both fruit set per flower and seed number per fruit. We found that both population size and the level of genetic relatedness among individuals influence maternal reproductive success. Our results explicitly define an ecological genetic obstacle faced by populations of an exotic species on its way to becoming invasive. Such a mechanistic understanding of the invasions of species that require a mate can and should be exploited for both controlling current outbreaks and reducing their frequency in the future.

California↗

PubMed2005

Accumulation of polycyclic aromatic hydrocarbons from creosote-contaminated soil in selected plants and the oligochaete worm Enchytraeus crypticus.

The accumulation of PAHs from a creosote-contaminated soil was examined in laboratory experiments using English ryegrass (Lolium perenne), white clover (Trifolium repens) and radish (Raphanus sativus), and the oligochaete worm Enchytraeus crypticus. Toxicity to the plants and the worms was assessed, and a soil sample mixed with calcined sand was used for accumulation experiments to avoid interference from toxicity in the soil. Accumulation of potentially carcinogenic PAHs varied among the plants, and there was a linear relation between concentrations of PAHs in the soil and in the plants. Correlations between values of the biota-soil accumulation factors and octanol-water partition coefficients, or water solubility varied among the plants and were rather weak, so that lipophilic character or water solubility of the PAHs alone cannot explain PAH accumulation. Accumulation of carcinogenic PAHs from the soil, in the presence of the other PAHs was greatest for Trifolium repens. PAHs were accumulated in the oligochaete worm (Enchytraeus crypticus), and biota-soil accumulation factors exceeded those for the plants. It is suggested that site-specific evaluation of contaminated sites should include not only chemical analysis and evaluation of toxicity but also accumulation of contaminants into biota such as plants and worms.

Animals↗

PubMed2004

Flower color microevolution in wild radish: evolutionary response to pollinator-mediated selection.

Evolutionary ecologists are fundamentally interested in how species interactions affect evolutionary change. We tested the degree to which plant-pollinator interactions affect the frequency of flower color morphs of Raphanus sativus. Petal color in R. sativus is determined by two independently assorting loci, producing four petal colors (yellow, white, pink, and bronze). We assessed the impact of pollinator discrimination on changes in flower color variation by comparing the frequency of colors produced in the presence (open pollination) versus absence (null pollination) of pollinator discrimination. We also assessed the impact of postpollination and developmental effects on progeny colors using equal pollinations with all four color morphs. Our results from open pollinations found an overrepresentation of yellow progeny in the next generation, when compared with both null pollinations and cumulative ratios based on Hardy-Weinberg and linkage equilibria assumptions. When these results were combined with those from equal pollinations, the overrepresentation of yellow could be attributed to selection from pollinators. Yet, surveys in the field the following year found no flower color frequency changes in the next generation. These results illustrate that flower color microevolution can be driven by both pollinator discrimination and other nonpollinator selective forces acting during the seed-to-adult transition, countering selection imposed by pollinators.

Animals↗

PubMed2004

Uptake and translocation of phytochemical 2-benzoxazolinone (BOA) in radish seeds and seedlings.

The molecular aspects of phytochemical interactions between plants, especially the process of phytochemical translocation by the target plant, remain challenging for those studying allelopathy. 2-Benzoxazolinone (BOA) is a natural chemical produced by rye (Secale cereale) and is known to have phytotoxic effects on weed seeds and seedlings. The translocation of BOA into target plants has been poorly investigated. Therefore, the total absorption of [ring U 14C] BOA was estimated by oxidizing whole seedlings of Raphanus sativus cv. for 8 days and quantifying the radioactivity. Non-radiolabelled BOA in seedlings was also estimated by HPLC. BOA applied at 10(-3) M was readily taken up by germinated radish at a rate of 1556 nmol g(-1) FW. At these same concentrations, BOA reduced radish germination by 50% and caused a delay in radicle elongation. Exogenous BOA was responsible for the observed germination inhibition. At a concentration of 10(-5) M, BOA was taken up by germinated seeds (31 nmol g(-1) FW), but this quantity did not affect radish germination. Labelled BOA was not mineralized in the culture medium during seedling growth as no 14CO2 was recovered. Both 10(-3) and 10(-5) M BOA were translocated into radish organs, mainly into roots and cotyledons. These organs were then identified as potential physiological target sites. Cotyledons remained the target sink (44% of the total radioactivity). The kinetics of BOA uptake at 10(-3) and 10(-5) M in radish seedlings was identical: BOA accumulation was proportional to its initial concentration. A comparison between radioactivity and HPLC quantification for 10(-3) M BOA indicated that BOA (along with some metabolites) could effectively be recovered in radish organs using chromatography.

Benzoxazoles↗

PubMed2004

Water channel activity of radish plasma membrane aquaporins heterologously expressed in yeast and their modification by site-directed mutagenesis.

Plants contain a number of aquaporin isoforms. We developed a method for determining the water channel activity of individual isoforms of aquaporin. Six plasma membrane aquaporins (RsPIPs) and two vacuolar membrane aquaporins (RsTIPs) of radish (Raphanus sativus) were expressed heterologously in Saccharomyces cerevisiae BJ5458, which is deficient in endogenous functional aquaporin. Aquaporins were detected by immunoblot analysis with corresponding antibodies. Water permeability of membranes from yeast transformants was assayed by stopped-flow spectrophotometry. The water channel activity of members of the RsPIP2 and RsTIP subfamilies was about 10 times and 5 times greater, respectively, than that of the control; however, RsPIP1s had little (RsPIP1-2 and RsPIP1-3) or no activity (RsPIP1-1). Site-directed mutation of several residues conserved in RsPIP1s or RsPIP2s markedly altered the water transport activity. Exchange of Ile244 of RsPIP1-3 with valine increased the activity to 250% of the wild type RsPIP1-3. On the other hand, exchange of Val235 of RsPIP2-2, which corresponds to RsPIP1-3 Ile244, with isoleucine caused a marked inactivation to 45% of the original RsPIP2-2. Mutation at possible phosphorylation sites at the N- and C-terminal tails also altered the activity. These results suggest that these residues in the half-helix loop E and the tails are involved in the water transport and the functional regulation of RsPIP1 and RsPIP2.

Amino Acid Sequence↗

PubMed2005

The zinc uptake regulator Zur is essential for the full virulence of Xanthomonas campestris pv. campestris.

Zur is a regulator of the high-affinity zinc uptake system in many bacteria. In Xanthomonas campestris pv. campestris 8004, a putative protein encoded by the open reading frame designated as XC1430 shows 42% amino acid similarity with the Zur of Escherichia coli. An XC1430-disrupted mutant 1430nk was constructed by homologous suicide plasmid integration. 1430nk failed to grow in rich medium supplemented with Zn2+ at a concentration of 400 microM and in nonrich medium supplemented with Zn2+ at a concentration of 110 microM, whereas the wild-type strain grew well in the same conditions. In rich medium with 400 microM Zn2+, 1430nk accumulated significantly more Zn2+ than the wild-type strain. 1430nk showed a reduction in virulence on the host plant Chinese radish (Raphanus sativus L. var. radiculus Pers.) and produced less extracellular polysaccharide (EPS) than did the wild-type strain in the absence of added zinc. These results revealed that XC1430 is a functional member of the Zur regulator family that controls zinc homeostasis, EPS production, and virulence in X. campestris pv. campestris.

Amino Acid Sequence↗

PubMed2003

Hydrotropism interacts with gravitropism by degrading amyloplasts in seedling roots of Arabidopsis and radish.

In response to a moisture gradient, roots exhibit hydrotropism to control the orientation of their growth. To exhibit hydrotropism, however, they must overcome the gravitropism that is dominant on Earth. We found that moisture gradient or water stress caused immediate degradation of the starch anchors, amyloplasts, in root columella cells of Arabidopsis and radish (Raphanus sativus). Namely, development of hydrotropic response was accompanied by a simultaneous reduction in starch content in columella cells. Rapid degradation of amyloplasts in columella cells also occurred in the water-stressed roots with sorbitol or mannitol. Both hydrotropically stimulated and water-stressed roots showed a reduced responsiveness to gravity. Roots of a starchless mutant, pgm1-1, showed an enhanced hydrotropism compared with that of the wild type. These results suggest that the reduced responsiveness to gravity is, at least in part, attributable to the degradation of amyloplasts in columella cells. Thus, the reduction in gravitropism allows the roots to exhibit hydrotropism.

Arabidopsis↗

PubMed2003

Structural and biochemical dissection of photorespiration in hybrids differing in genome constitution between Diplotaxis tenuifolia (C3-C4) and radish (C3).

We compared the structural, biochemical, and physiological characteristics involved in photorespiration of intergeneric hybrids differing in genome constitution (DtDtR, DtDtRR, and DtRR) between the C(3)-C(4) intermediate species Diplotaxis tenuifolia (DtDt) and the C(3) species radish (Raphanus sativus; RR). The bundle sheath (BS) cells in D. tenuifolia included many centripetally located chloroplasts and mitochondria, but those of radish had only a few chloroplasts and mitochondria. In the hybrids, the numbers of chloroplasts and mitochondria, the ratio of centripetally located organelles to total organelles, and the mitochondrial size in the BS cells increased with an increase in the constitution ratio of the Dt:R genome. The P-protein of glycine decarboxylase (GDC) was confined to the BS mitochondria in D. tenuifolia, whereas in radish, it accumulated more densely in the mesophyll than in the BS mitochondria. In the hybrids, more intense accumulation of GDC in the BS relative to the mesophyll mitochondria occurred with an increase in the Dt:R ratio. These structural and biochemical features in the hybrids were reflected in the gas exchange characteristics of leaves, such as the CO(2) compensation point. Our data indicate that the leaf structure, the intercellular pattern of GDC expression, and the gas exchange characteristics of C(3)-C(4) intermediate photosynthesis are inherited in the hybrids depending on the constitution ratio of the parent genomes. Our findings also demonstrate that the apparent reduced photorespiration in C(3)-C(4) intermediate plants is mainly due to the structural differentiation of mitochondria and chloroplasts in the BS cells combined with the BS-dominant expression of GDC.

Amino Acid Oxidoreductases↗

PubMed2005

Molecular cloning of a {beta}-galactosidase from radish that specifically hydrolyzes {beta}-(1->3)- and {beta}-(1->6)-galactosyl residues of Arabinogalactan protein.

A basic beta-galactosidase with high specificity toward beta-(1-->3)- and beta-(1-->6)-galactosyl residues was cloned from radish (Raphanus sativus) plants by reverse transcription-PCR. The gene, designated RsBGAL1, contained an open reading frame consisting of 2,532 bp (851 amino acids). It is expressed in hypocotyls and young leaves. RsBGAL1 was highly similar to beta-galactosidases having exo-beta-(1-->4)-galactanase activity found in higher plants and belongs to family 35 of the glycosyl hydrolases. Recombinant RsBGAL1 was expressed in Pichia pastoris and purified to homogeneity. The recombinant enzyme specifically hydrolyzed beta-(1-->3)- and beta-(1-->6)-galactooligosaccharides, the same substrates as the native enzyme isolated from radish seeds (Sekimata et al., 1989). It split off about 90% of the carbohydrate moieties of an arabinogalactan protein extracted from radish roots in concerted action with microbial alpha-l-arabinofuranosidase and beta-glucuronidase. These results suggest that RsBGAL1 is a new kind of beta-galactosidase with different substrate specificity than other beta-galactosidases that exhibit exo-beta-(1-->4)-galactanase activity. The C-terminal region (9.6 kD) of RsBGAL1 is significantly similar to the Gal lectin-like domain, but this region is not retained in the native enzyme. Assuming posttranslational processing of RsBGAL1 with elimination of the Gal lectin-like domain results in a protein consisting of two subunits with molecular masses of 46 and 34 kD (calculated from the RsBGAL1 gene sequence). This is in good agreement with the SDS-PAGE and matrix-assisted laser desorption/ionization-time-of flight mass spectrometry measurements for subunits of the native enzyme (45 and 34 kD) and may thus partially explain the formation process of the native enzyme.

Amino Acid Sequence↗

PubMed2005

Characterization and cloning of the chlorophyll-degrading enzyme pheophorbidase from cotyledons of radish.

Enzymatic removal of the methoxycarbonyl group of pheophorbide (Pheid) a in chlorophyll degradation was investigated in cotyledons of radish (Raphanus sativus). The enzyme pheophorbidase (PPD) catalyzes the conversion of Pheid a to a precursor of pyropheophorbide (PyroPheid), C-13(2)-carboxylPyroPheid a, by demethylation, and then the precursor is decarboxylated nonenzymatically to yield PyroPheid a. PPD activity sharply increased with the progression of senescence in radish, suggesting de novo synthesis of PPD. The enzyme activity was separated into two peaks in anion-exchange and hydrophobic chromatography; the terms type 1 and type 2 were applied according to the order of elution of these enzymes in anion-exchange chromatography. PPD types 1 and 2 were purified 9,999- and 6,476-fold, with a yield of 0.703% and 2.73%, respectively. Among 12 substrates tested, both enzymes were extremely specific for Pheids of the dihydroporphyrin and tetrahydroporphyrin types, indicating that they are responsible for the formation of these PyroPheids. Both PPDs had molecular masses of 113,000 kD on gel filtration and showed three bands of 16.8, 15.9, and 11.8 kD by SDS-PAGE. The partial N-terminal amino acid sequences for these bands of PPD (type 2) were determined. Based on their N-terminal amino acid sequences, a full-length cDNA of PPD was cloned. The molecular structure of PPD, particularly the molecular mass and subunit structure, is discussed in relation to the results of SDS-PAGE.

Amino Acid Sequence↗

PubMed2002

Reducing environmental bias when measuring natural selection.

Crucial to understanding the process of natural selection is characterizing phenotypic selection. Measures of phenotypic selection can be biased by environmental variation among individuals that causes a spurious correlation between a trait and fitness. One solution is analyzing genotypic data, rather than phenotypic data. Genotypic data, however, are difficult to gather, can be gathered from few species, and typically have low statistical power. Environmental correlations may act through traits other than through fitness itself. A path analytic framework, which includes measures of such traits, may reduce environmental bias in estimates of selection coefficients. We tested the efficacy of path analysis to reduce bias by re-analyzing three experiments where both phenotypic and genotypic data were available. All three consisted of plant species (Impatiens capensis, Arabidopsis thaliana, and Raphanus sativus) grown in experimental plots or the greenhouse. We found that selection coefficients estimated by path analysis using phenotypic data were highly correlated with those based on genotypic data with little systematic bias in estimating the strength of selection. Although not a panacea, using path analysis can substantially reduce environmental biases in estimates of selection coefficients. Such confidence in phenotypic selection estimates is critical for progress in the study of natural selection.

Arabidopsis↗

PubMed2003

Patterns and mechanisms of selection on a family-diagnostic trait: evidence from experimental manipulation and lifetime fitness selection gradients.

Plant traits that show little variation across higher taxa are often used as diagnostic traits, but the reason for the stasis of such traits remains unclear. Wild radish, Raphanus raphanistrum, exhibits tetradynamous stamens (four long and two short, producing a dimorphism in anther height within each flower), as do the vast majority of the more than 3,000 species in the Brassicaceae. Here we examine the hypothesis that selection maintains the stasis of dimorphic anther height by investigating the effects of this trait on pollen removal, seed siring success, and seed set in R. raphanistrum using both experimental and observational methods. Observational selection gradient analysis based on lifetime seed siring success provided evidence for an optimum dimorphism that was greater than zero in one of three years. In both experimentally manipulated and unmanipulated flowers, more pollen was removed in single visits from flowers with less dimorphism. There was no significant effect of anther dimorphism on female fitness (seed set). Therefore, there is some evidence to suggest that selection is maintaining anther dimorphism in wild radish, and that higher male fitness might result from restriction of single-visit pollen removal. We discuss these results in light of pollen presentation theory.

Flowers↗

PubMed2006

Isolation and characterization of rhizosphere bacteria with potential for biological control of weeds in vineyards.

AIMS: Deleterious rhizosphere inhabiting bacteria (DRB) have potential to suppress plant growth. This project focuses on the isolation of DRB with potential for development as commercial products for weed control. METHODS AND RESULTS: Bacteria were isolated from the rhizosphere, rhizoplane, and endorhizosphere of seedlings and mature plants of wild radish (Raphanus raphanistrum), annual ryegrass (Lolium rigidum) and capeweed (Arctotheca calendula) growing in vineyards in the Swan Valley, Western Australia. A majority (81.5%) of the 442 strains was obtained from either rhizospheres or rhizoplanes. Rapid screening techniques were developed to evaluate in the laboratory and glasshouse the effects of bacteria on plants. Strains were screened in the glasshouse for deleterious effects on annual ryegrass, wild radish, grapevine rootlings (Vitis vinifera) and the legume cover crop subterranean clover (Trifolium subterraneum). Three strains were identified using the Biolog system and 16S rRNA gene sequencing as two strains of Pseudomonas fluorescens (WSM3455 and WSM3456) and one strain of Alcaligenes xylosoxidans (WSM3457). One of the P. fluorescens (WSM3455) strain produced hydrogen cyanide, an inhibitor of plant roots and a broad-spectrum antimicrobial compound. CONCLUSIONS: Three strains specifically inhibited wild radish but had no significant deleterious effects on either grapevine rootlings or subterranean clover. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggested manipulation of the weed seedling rhizosphere using identified DRB as a potential biocontrol agent for wild radish.

Bacteria↗

PubMed2006

Radish seed extract mediates its cardiovascular inhibitory effects via muscarinic receptor activation.

In this study, we describe the hypotensive, cardio-modulatory and endothelium-dependent vasodilator actions of Raphanus sativus (radish) seed crude extract in an attempt to provide scientific basis for its traditional use in hypertension. The plant extract (Rs.Cr) was prepared in distilled water and was subjected to phytochemical screening using standard analytical procedures. In vivo blood pressure was monitored in anaesthetized normotensive rats. Isolated tissue preparations were suspended in tissue baths containing Kreb's solution while acute toxicity study was performed in mice for 24 h. Rs.Cr tested positive for the presence of saponins, flavonoids, tannins, phenols and alkaloids and caused a dose-dependent (0.1-3 mg/kg) fall in blood pressure and heart rate of rats that was mediated via an atropine-sensitive pathway. In isolated guinea-pig atria, Rs.Cr showed dose-dependent (0.03-3.0 mg/mL) inhibition of force and rate of contractions. In the atropine-treated tissues, the inhibitory effect was abolished and a cardiac stimulant effect was unmasked which was resistant to adrenergic and serotonergic receptor blockade. In the endothelium-intact rat aorta, Rs.Cr inhibited phenylephrine-induced contractions, which was blocked by atropine and Nomega-Nitro-L-arginine methyl ester hydrochloride while was also absent in the endothelium-denuded preparations. The extract was safe in mice up to the dose of 10 g/kg. The study shows that the cardiovascular inhibitory effects of the plant are mediated through activation of muscarinic receptors thus possibly justifying its use in hypertension.

Animals↗