[Influence of calcium and rhizobial infections (Rhizobium leguminosarum) on the dynamics of nitric oxide (NO) content in roots of etiolated pea (Pisum sativum L.) seedlings].

The effect of exogenous calcium (Ca2+) and rhizobial infections (Rhizobium leguminosarum bv viceae) on the dynamics of the level of nitric oxide (NO) was studied in cross cuts of roots of two-day-old etiolated pea seedlings (Pisum sativum L.) using a DAF-2DA fluorescent probe. Fluctuations of the NO level, indicating the presence of a rhythm in the generation of NO in roots, were observed during the incubation of seedlings in water, a CaCl2 solution, and with rhizobial infections. Exogenous factors (Ca2+ and two rhizobial stamms) change the time dynamics of the NO level in comparison with the control (water).

[Influence of rhizobial (Rhizobium leguminosarum) inoculation and calcium ions on the NADPH oxidase activity in roots of etiolated pea (Pisum sativum L.) seedlings].

Changes in the functional activity of the NADPH oxidase in the microsomal fraction of roots of etiolated pea seedlings, caused by rhizobial inoculation and calcium ions (Ca2+), are shown. The enzyme activity in a medium with an exogenous source of Ca2+ (CaCl2, 100 microM) fluctuated, increasing 5 to 20 min and decreasing 10 and 30 min after addition. A calcium chelator (ethylene glycol tetraacetic acid (EDTA), 100 microM) potentiated the decrease in the enzyme activity in the presence of exogenous calcium. Rhizobial inoculation caused a 3.9-fold increase in the enzyme activity 5 min after inoculation compared to the control (without inoculation). The Ca(2+)-channel activator (amiodarone, 300 microM) and the Ca(2+)-channel blocker (lanthanum chloride, 400 microM) reduced the NADPH oxidase activity after rhizobial inoculation compared to the control level (without inoculation). It is concluded that Ca2+ and reactive oxygen species are involved in the regulation of the membrane NADPH oxidase activity in roots of pea seedlings.

[Level nitric oxide (NO) and growth of roots of etiolated pea seedlings].

Data regarding the interrelation of nitric oxide (NO) content in roots of 3-day-old etiolated pea seedlings and their growth under different concentrations of N-containing compounds were obtained. The concentration of exogenous compounds (sodium nitroprusside SNP, KNO3, NaNO2, L-arginine) rendering an inhibiting effect on the growth of roots were established, and the NO content in roots was determined at these concentration. It was shown that the inhibition of growth and highest NO content in the roots was determined with SNP (4 mM) and NaNO2 (2 mM) during 24 h exposition of seedlings. This dependence was not established in combinations with KNO3 (20 mM) and L-arginine (4 mM). We established that a NO scavenger, hemoglobin (4 µM), fully or partially removed the toxic effect of SNP, nitrate, and nitrite on growth. The effect of NO on the growth and the participation of N-containing compounds in generation of NO in roots of pea seedlings is discussed.

[Influence of environmental factors on the generation of nitric oxide in the roots of etiolated pea seedlings].

The article studies the nitric oxide (NO) levels in the roots of etiolated seedlings of garden peas (Pisum sativum L.) using the DAF-2DA fluorescent probe and fluorescent microscopy. Cross sections of roots of 100-150 microm (the site of a root which is 10-15 mm from the apex) are analyzed. It is shown that the level of NO in the roots after 24 h increased by more than a factor of 2 in the versions with NaNO2 and sodium nitroprusside. At feeding the seedlings with KNO3, a peak in the accumulation of NO in the roots (twofold increase) was observed after 30 min. Fertilizing seedlings with L-arginine (2 mM) increased the intensity of the fluorescence of the root sections by more than a factor of 2. The inoculation of seedlings of rhizobia (Rhizobium leguminosarum by. viceae) contributed to the reduction of NO on the background of the control (H20) and sodium nitroprusside and nitrogen compounds. Scavengers of NO (2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), hemoglobin) and inhibitors of nitrate reductase and animal NO synthase (sodium tungstate and aminoguanidine hydrochloride) reduced the level of NO in the roots. The results are discussed in relation to the role of NO in plants under the influence of biotic and abiotic factors.

[Structural and functional characteristics of plant NADPH oxidase: a review].

Data on structural and functional characteristics of plant NADPH oxidase (Rboh) are generalized. The enzyme homologs identical to the subunit gp91(phox) of the enzymatic complex of animal cells were found in plants. The activation of Rboh depends on the influx of Ca2+ into the cytoplasm and phosphorylation of the N-terminal region of the enzyme by Ca(2+)-dependent protein kinase. The possibility of the involvement of Rop GTPase, a cytosolic component of Rboh, in the activation of Rboh is discussed. It is postulated that Rboh localizes on the plasma membrane of plant cells. Rboh is activated under the influence of both biotic and abiotic factors, which is apparently associated with Ca2+ fluxes, reactive oxygen and nitrogen species, and transduction of information to the nuclear genome.

[The NADPH oxidase activity of pea seedling roots in rhizobial infection depending on abiotic and biotic factors].

The changes in NADPH activity was studied in the roots of 3-4-day-old etiolated pea (cultivar Aksaiskii usatyi) seedlings depending on plant inoculation with Rhizobium leguminosarum bv. viceae (strain CIAM 1026), adverse environmental factors (low temperature and high dose of a mineral nitrogen fertilizer), chemical substances (sodium nitroprusside and methyl viologen, or paraquat), and a biotic factor--the bacterium Escherichia coli (strain XL-1 Blue). It was demonstrated that all exogenous factors increased the activity of microsomal NADPH oxidase. Rhizobial infection removed the activation caused by exogenous factors only in the case of high nitrogen content in the medium, thereby displaying an antagonistic effect. A synergistic action on the enzyme activity was observed in the variants with combined action of rhizobia + paraquat and rhizobia + E. coli. An increased NADPH oxidase activity coincided with a growth inhibition of pea seedling roots. The results are discussed from the standpoint of the roles of NADPH oxidase and reactive oxygen species in the legume-rhizobium symbiosis.

[Reactive oxygen and nitrogen species in legume-rhizobial symbiosis: a review].

Published data on the role of reactive oxygen and nitrogen species (ROS and RNS, respectively) in the formation and functioning of the legume-rhizobial symbiosis are summarized. It is assumed that ROS and RNS fulfill a double function in the legume-rhizobial symbiosis by triggering the mechanisms enabling symbiosis formation and the mechanisms preventing the development of symbiotic structures (i.e., the defensive responses). A hypothetic scheme illustrating the involvement of ROS and RNS in the formation of legume-rhizobial symbiosis is proposed.

[The influence of mineral nitrogen on legume-rhizobium symbiosis].

A literature review synthesizes the data on physiological mechanisms of the influence of high doses of mineral nitrogen (nitrates and ammonium) on the formation and functioning of legume-rhizobium symbiosis. The participation of phytohormonal and phenolic metabolism and active forms of oxygen and nitrogen in the symbiosis is highlighted. Close connection between these metabolic processes in the formation and functioning of legume-rhizobium symbiosis under a redundant supply of plants by mineral nitrogen is underlined.

[Effect of nitric oxide and other nitrogen compounds on the adhesion and penetration of nodule bacteria into root tissues and on growth of etiolated pea seedlings].

The action of sodium nitroprusside, a nitric oxide donor, and other nitrogen compounds (KNO3, KNO2, and (NH4)2SO4) on adhesion and penetration of nodule bacteria into root tissues of etiolated pea seedlings was studied. Only nitroprusside displayed a clearly negative effect on rhizobium adhesion and penetration and seedling growth. This effect was not observed with other nitrogen compounds even at high (20 mM) concentrations. Hemoglobin attenuated the negative effect of nitroprusside on bacteria and seedlings. The results are discussed in the context of the role of nitric oxide in the life of plants and nodule bacteria.

[Oxidation processes at initial stages of interaction of root nodule bacteria (Rhizobium leguminosarum) and pea (Pisum sativum L.): a review].

A possible physiological mechanism of legume-Rhizobium symbiosis, consisting in regulation of the intensity of oxidation processes by the microsymbiont in response to infection with Rhizobium, was analyzed using our own and published data. The results used in the analysis included data on the content of reactive oxygen species (O2*-, and H2O2), activity of antioxidant enzymes (superoxide dismutase, catalase, and peroxidase), and intensity of lipid peroxidation proceeding with the involvement of lipophilic phenolic compounds of the microsymbiont.

[The defense and regulatory mechanisms during development of legume-Rhizobium symbiosis].

The roles of indolylacetic acid, the peroxidase system, catalase, active oxygen species, and phenolic compounds in the physiological and biochemical mechanisms involved in the autoregulation of nodulation in the developing legume-Rhizobium symbiosis were studied. It was inferred that the concentration of indolylacetic acid in the roots of inoculated plants, controlled by the enzymes of the peroxidase complex, is the signal permitting or limiting nodulation at the initial stages of symbiotic interaction. Presumably, the change in the level of active oxygen species is determined by an antioxidant activity of phenolic compounds. During the development of symbiosis, phytohormones, antioxidant enzymes, and active oxygen species may be involved in the regulation of infection via both a direct antibacterial action and regulation of functional activity of the host plant defense systems.

[Active oxygen species in pea seedlings during the interactions with symbiotic and pathogenic microorganisms].

The level of active oxygen species (AOS)--superoxide anion radical (O2*-) and hydrogen peroxide (H2O2)--in pea (Pisum sativum L.) cultivar Marat seedlings was studied upon their inoculation with symbiotic (Rhizobium leguminosarum bv. viceae strain CIAM 1026) and pathogenic (Pseudomonas syringae pv. pisi Sackett) microorganisms. Different patterns of the changes in AOS in pea seedlings during the interactions with the symbiont and the phytopathogen were recorded. It is assumed that O2*- and H2O2 are involved in the defense and regulatory mechanisms of the host plant.

[Hydrogen peroxide content and catalase activity at inoculation with root tubercle bacteria of pea seedlings with the various nodulation ability].

Hydrogen peroxide (H2O2) content and catalase activity were studied in pea (Pisum sativum L.) seedlings with normal (cultivar Marat) and disrupted (pea mutants) process of nodulation, which were inoculated with the nitrogen-fixing bacterium Rhizobium leguminosarum strain CIAM 1026. Differences in hydrogen peroxide content and catalase activity in pea seedlings with different ability for nodulation, which were inoculated with rhizobia, were found. It was assumed that H2O2 and catalase are involved in defensive and regulatory mechanisms in the host plant.

[Possible involvement of hydrogen peroxide and salicylic acid in the legume-rhizobium symbiosis].

H2O2 content was studied in the roots and epicotyls of pea (Pisum sativum L.) with normal (cultivar Marat) and disturbed (non-nodulating mutant K14 and hypernodulating mutant Nod3) regulation of root nodulation after inoculation with active industrial strain of Rhizobium leguminosarum by. viceae 250a/CIAM 1026. Pea biotypes differed by H2O2 content in the roots and epicotyls. Exogenous salicylic acid (SA) (0.2 mM) affected H2O2 and SA contents in the roots in an inoculation-dependent manner. The involvement of hydrogen peroxide and SA as signaling molecules as well as of antibacterial agents in the pea-rhizobium interaction at the initial stages of symbiosis is proposed.