scholarly journals Activity of nitrogen fixation and antioxidant enzymes in symbiotic systems Glycine max – Bradyrhizobium japonicum for complex treatment with lectin and fungicides

2018 ◽  
Vol 9 (2) ◽  
pp. 148-155
Author(s):  
S. Y. Kots ◽  
T. P. Mamenko ◽  
A. V. Pavlyshche
Keyword(s):  
Superoxide Dismutase ◽  
Nitrogen Fixation ◽  
Antioxidant Enzymes ◽  
Bradyrhizobium Japonicum ◽  
Root Nodules ◽  
Guaiacol Peroxidase ◽  
Complex Treatment ◽  
Mass Flowering ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity

The dynamics of the nitrogen fixation activity of the root nodules, the growth of the vegetative mass of plants and the change in the activity of antioxidant enzymes (superoxide dismutase, ascorbate and guaiacol peroxidase) in different soybean organs for treatment of seeds by rhizobia incubated with lectin, in combination with fungicides have been studied. The objects of the study were symbiotic systems formed with the participation of soybean (Glycine max (L.) Merr.) Almaz and Bradyrhizobium japonicum (standard strain 634b) incubated with lectin. As disinfectants of soybean seeds, the following preparations with fungicidal activity were used – Maxim XL 035 PS, Fever, Standak Top according to one rate of active substance consumption of each preparation specified by the manufacturer. One part of the seeds treated with fungicides was inoculated with pure culture of suspension of rhizobia for one hour (titre of suspension concentration was 108 cells/ml). Another part of the seeds treated with fungicides was inoculated with rhizobia suspension, which was previously incubated with a solution of commercial lectin soybean at a concentration of 100 μg/ml. The research was conducted in strictly controlled conditions of a model vegetative experiment using microbiological, physiological, biochemical methods, gas chromatography, spectrophotometry. It was found that processing of soybean seeds with fungicides (Fever and Maxim XL) together with rhizobium inoculation contributed to the preservation of the nitrogen fixation activity of the root nodules and the growth of vegetative mass of plants. Under these conditions, the intensification of the activity of superoxide dismutase and ascorbate peroxidase was observed, as well as inhibition of the activity of guaiacol peroxidase in soybean root nodules in the phase of three true leaves and increased activity of all investigated enzymes in the phase of mass flowering. It has been established that the use of complex treatment of seeds by soybean rhizobia incubated with lectin and fungicides leads to an increase in the activity of superoxide dismutase and guaiacol peroxidase in root nodules in the phase of three true leaves and the growth of the activity of ascorbate peroxidase in the phase of mass flowering. At the same time, the inhibition of the growth of vegetative mass of plants and their symbiotic properties occurred, as evidenced by the decrease in the nitrogen fixation activity of the root nodules for the joint treatment of seeds with fungicides and lectin. A specific reaction of investigated enzymes in the roots and leaves of soybean was shown, which was more pronounced in the phase of three true leaves, indicating the development of a typical antioxidant reaction to a complex treatment, as a kind of stress that is leveled to the phase of mass flowering. The degree of reaction of antioxidant enzymes in the studied symbiotic systems Glycine max – Bradyrhizobium japonicum depends on the nature of the active substance fungicides and the manifestation of their joint effect in a complex with rhizobia incubated with lectin.

scholarly journals The use of Tn5-mutants Bradyrhyzobium japonicum for regulation of prooxidant-antioxidant processes in soybean plants under drought condition

2020 ◽  
Vol 27 ◽  
pp. 119-124
Author(s):  
S. Ya. Kots ◽  
T. P. Mamenko ◽  
R. A. Yakymchuk
Keyword(s):  
Hydrogen Peroxide ◽  
Nitrogen Fixation ◽  
Bradyrhizobium Japonicum ◽  
Prolonged Exposure ◽  
Root Nodules ◽  
Soybean Seed ◽  
Crop Productivity ◽  
Guaiacol Peroxidase ◽  
Negative Effects ◽  
Nitrogen Fixation Activity

Aim. To investigate the use of soybean seed inoculation by nodule bacteria obtained by transposon mutagenesis to ensure the effective formation and functioning of symbiotic systems by regulating prooxidant - antioxidant processes and reducing the negative effects of drought on crop productivity. Methods. Microbiological, physiological, biochemical methods, gas chromatography and spectrophotometry. Results. It has been proved that due to the activation of protective antioxidant enzymes of catalase, ascorbate and guaiacol peroxidase in soybean roots and root nodules, adaptive rearrangements of plant metabolism occur aimed at stabilizing the content of prooxidants, hydrogen peroxide, in drought conditions. At the same time, the specific nitrogen-fixation activity of soybean root nodules undergoes no significant changes and indicates the preservation of the effective functioning of the symbiotic apparatus, is the result of activation of protective antioxidant processes and adaptation of the soybean symbiotic system with the participation of Tn-5 mutant Bradyrhizobium japonicum B1-20 to dehydration conditions. Conclusions. The use of inoculation of soybean seeds with the Tn-5 mutant Bradyrhizobium japonicum B1-20 leads to regulation of prooxidant - antioxidant protective processes in plants, helps to increase their nitrogen-fixation potential and maintain grain yield under prolonged exposure to drought. Keywords: soybean (Glycine max (L.) Merr.), hydrogen peroxide, catalase, ascorbate peroxidase, guaiacol peroxidase, drought.

scholarly journals The reaction of self-fertile alfalfa lines to inoculation with nodule bacteria

2020 ◽  
Keyword(s):  
Nitrogen Fixation ◽  
Sinorhizobium Meliloti ◽  
Root Nodules ◽  
Growing Season ◽  
Dry Mass ◽  
Nodule Bacteria ◽  
Forage Crop ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity ◽  
Different Strains

Under the conditions of a model pot experiments, the reaction of the self-fertile lines of alfalfa Kishvardy 46, Kishvardy 27, Vertus and Ziguen to inoculation with nodule bacteria Sinorhizobium meliloti AC48 and AC88 was studied. As a result of studies, it was found that the intensity of assimilation of N2 by symbiotic systems created with the participation of various genotypes of alfalfa and active strains of S. meliloti is one of the main factors that affects the vegetative mass yield of this important forage crop. Self-fertile lines of Medicago sativa L. plants, inoculated with different strains of rhizobia were characterized by higher rates of the mass formed on the root nodules, compared to the control plants of the alfalfa variety Yaroslavna. The traditional dynamics of nitrogen-fixation activity of root nodules was maintained in all the symbiotic systems studied by us, with low values in the stems formation stage and intensive growth in the budding and flowering stages. The highest level of nitrogen fixation and vegetative growth of plants (values of plants green and dry mass, roots and root nodules mass) was established by inoculation of alfalfa line Kishvardy 46 with strain S. meliloti AC48. During the growing season the indices of the mass of nodules formed on the roots of these plants were higher by 1.8–2.3 times, the green mass by 1.2–1.6 times and the height of the plants 1.2–1.4 times as compared to the control. In the flowering stages the nitrogen-fixation activity of the symbiotic complex of plants of the Kishvardy line 27 and nodule bacteria S. meliloti AC48 exceeded the values in the symbiotic systems formed with the participation of the same strain and plants of the Ziguen and Vertus lines by 13.0 and 39.4 %. The lowest values of nitrogen fixation activity were observed by inoculation of plants of the Vertus and Ziguen lines with active strains S. meliloti AC48 and AC88 compared to the symbioses formed by the plants of the Kishvardy lines 27 and 46, as well as of the control-variety Yaroslavna with the noted strains. A stimulating effect of inoculation of alfalfa seeds of different genotypes on the growth and development of plants was noted, as evidenced by the positive dynamics of the increase in above-ground mass, the accumulation of dry matter and higher than the control values (indicators) of plant height during the growing season.

Confirmation of the infectivity of a free-living actinomycete isolated from Comptonia peregrina root nodules by immunological and ultrastructural studies

1978 ◽  
Vol 56 (21) ◽  
pp. 2621-2635 ◽  
Author(s):  
Maurice Lalonde
Keyword(s):  
Nitrogen Fixation ◽  
Pure Culture ◽  
Root Nodules ◽  
Free Living ◽  
Ultrastructural Studies ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity ◽  
Membrane Envelope ◽  
Freeze Etching

The inoculation of the host plant Comptonia peregrina (L.) Coult. (Myrica asplenifolia L.) by a pure culture of a free-living actinomycete, isolated from Comptonia root nodules by Callaham et al. (1978, Science, 199: 899–902), was successful. Short-term and long-term nodulation tests confirmed the infectivity of the Comptonia isolate. Acetylene reduction assays of the nodules induced by this prokaryote isolate demonstrated nitrogen fixation activity. This nitrogen fixation activity was able to sustain a prolific growth of the nodulated host plants growing in a N-free substrate. Indirect immufluorescence reactions, using specific gamma globulin against the actinomycetal isolate of the Comptonia root nodule, demonstrated the identity of this actinomycete in these in vitro produced Comptonia root nodules. Ultrastructure of the Comptonia isolate, developing as a free-living or endophytic actinomycete, was observed by light microscopy, freeze etching, and transmission electron microscopy. The free-living filamentous and sporulating isolate produced typical hyphae and vesicles when growing as an endophyte in the Comptonia nodule. These endophytic hyphae and vesicles were always encapsulated by a polysaccharide material which was surrounded by a host membrane envelope. A polysaccharide capsule was not demonstrated on the free-living Comptonia isolate. The endophytic vesicles were club shaped and highly septate. Such vesicles were never produced by the free-living isolate growing in an artificial medium. The Comptonia isolate is a spore former in pure culture and was able to sporulate in basal tissues of 5-month-old Comptonia nodules.

scholarly journals Effect of abscisic acid on symbiotic nitrogen fixation activity in the root nodules ofLotus japonicus

2010 ◽  
Vol 5 (4) ◽  
pp. 440-443 ◽  
Author(s):  
Akiyoshi Tominaga ◽  
Maki Nagata ◽  
Koichi Futsuki ◽  
Hidetoshi Abe ◽  
Toshiki Uchiumi ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Abscisic Acid ◽  
Symbiotic Nitrogen Fixation ◽  
Root Nodules ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity

scholarly journals PERSPECTIVE USE OF TN5-MUTANTS OF RHIZOBIA IN PREPARATION OF BACTERIAL FERTILIZERS

2009 ◽  
Vol 8 ◽  
pp. 32-39
Author(s):  
S.Ya. Kots ◽  
S.M. Malychenko ◽  
P.M. Mamenko ◽  
G.M. Drozdenko
Keyword(s):  
Escherichia Coli ◽  
Nitrogen Fixation ◽  
Bradyrhizobium Japonicum ◽  
Protein Composition ◽  
Transposon Mutagenesis ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity ◽  
Tn5 Mutants

By using strain of Escherichia coli with build-in plasmid pSUP2021::Tn5 the transposon mutagenesis of Bradyrhizobium japonicum (strains 646, 614a and 71t) was performed. The highest formation frequency of kanamycinresistant mutants was 10-6 (B. japonicum 646). Among the 1500 mutants obtained, the best were selected by the “virulence”, “nitrogen fixation activity” and “symbiosis efficiency” criteria. In spite of different symbiotic characteristics of the selected Tn-5 mutants no considerable differences in their protein composition were determined. The authors made conclusion on the possibility of use of transposon mutagenesis in order to obtain the new forms of slowgrowing bacteria.

scholarly journals The Bradyrhizobium japonicum Proline Biosynthesis Gene proC Is Essential for Symbiosis

2000 ◽  
Vol 66 (12) ◽  
pp. 5469-5471 ◽  
Author(s):  
Natalie D. King ◽  
David Hojnacki ◽  
Mark R. O'Brian
Keyword(s):  
Energy Source ◽  
Nitrogen Fixation ◽  
Mutant Strain ◽  
Bradyrhizobium Japonicum ◽  
Root Nodules ◽  
Proline Biosynthesis ◽  
Plant Host ◽  
Nitrogen Fixation Activity ◽  
Biosynthesis Gene ◽  
Exogenous Supply

ABSTRACT Plant host-derived proline is proposed to serve as an energy source for rhizobia in the rhizosphere and in symbiotic root nodules. TheBradyrhizobium japonicum proC gene was isolated, and aproC mutant strain that behaved as a strict proline auxotroph in culture was constructed. The proC strain elicited undeveloped nodules on soybeans that lacked nitrogen fixation activity and plant hemoglobin. We conclude that the proCgene is essential for symbiosis and suggest that the mutant does not obtain an exogenous supply of proline in association with soybeans sufficient to satisfy its auxotrophy.

Novel rhizobia exhibit superior nodulation and biological nitrogen fixation even under high nitrate concentrations

2019 ◽  
Vol 96 (2) ◽  
Author(s):  
Hien P Nguyen ◽  
Hiroki Miwa ◽  
Jennifer Obirih-Opareh ◽  
Takuya Suzaki ◽  
Michiko Yasuda ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Bradyrhizobium Japonicum ◽  
Root Nodules ◽  
Nitrogen Fertilizers ◽  
Nodule Formation ◽  
Nitrogen Fixing ◽  
Nitrogen Fixation Activity ◽  
Nitrate Concentrations ◽  
Nodulation Competitiveness ◽  
Biological Nitrogen

ABSTRACT Legume–rhizobium symbiosis leads to the formation of nitrogen-fixing root nodules. However, externally applied chemical nitrogen fertilizers (nitrate and ammonia) strongly inhibit nodule formation and nitrogen fixation. Here, we isolated several rhizobial strains exhibiting a superior nodulation and nitrogen fixation with soybean at high nitrate concentrations. The nodulation of soybean symbiont Bradyrhizobium diazoefficiens USDA110 was significantly inhibited at 12.5 mM nitrate; however, three isolates (NKS4, NKM2 and NKTG2) were capable of forming nitrogen-fixing nodules, even at 20 mM nitrate. These isolates exhibited higher nodulation competitiveness and induced larger nodules with higher nitrogen-fixation activity than USDA110 at 5 mM nitrate. Furthermore, these isolates induced more nodules than USDA110 even in nitrate-free conditions. These isolates had a distant lineage within the Bradyrhizobium genus; though they were relatively phylogenetically close to Bradyrhizobium japonicum, their morphological and growth characteristics were significantly different. Notably, in the presence of nitrate, expression of the soybean symbiosis-related genes (GmENOD40 and GmNIN) was significantly higher and expression of GmNIC1 that is involved in nitrate-dependent nodulation inhibition was lower in the roots inoculated with these isolates in contrast with inoculation of USDA110. These novel rhizobia serve as promising inoculants for soybeans cultivated in diverse agroecosystems, particularly on nitrate-applied soils.

Cereals frost resistance correlates with an integral indicators of content of low-molecular-weight protective compounds and activity of antioxidant enzymes

2020 ◽  
Vol 2020 (3) ◽  
pp. 71-86
Author(s):  
O. I. Horielova ◽  
◽  
N. I. Ryabchun ◽  
M. A. Shkliarevskyi ◽  
A. M. Reznik ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Superoxide Dismutase ◽  
Antioxidant Enzymes ◽  
Antioxidant System ◽  
Frost Resistance ◽  
Low Molecular Weight ◽  
Guaiacol Peroxidase ◽  
Wheat Seedlings ◽  
Enzymatic Antioxidant ◽  
Adult Plants

Along with specific adaptive reactions, universal defense reactions, in particular activation of antioxidant system, are of great importance for plant survival under cold conditions. We have studied a relationship among the content of low-molecular-weight protective compounds with antioxidant properties (proline, soluble carbohydrates, flavonoids), the activity of antioxidant enzymes (superoxide dismutase, catalase, and guaiacol peroxidase) in seedlings of winter wheat, rye and triticale, and frost resistance of etiolated seedlings and adult plants at tillering stage. It was found that there was a fairly close correlation between the frost resistance of seedlings and adult cereal plants (r = 0,78). It was shown that a pronounced relationship between individual indicators of antioxidant system functioning in unhardened seedlings and their frost resistance was not found. After 6-day hardening of seedlings at 2-4°C, there was a high correlation between the total indicator of the enzymatic antioxidant system (the sum of normalized indicators of superoxide dismutase, peroxidase, and catalase activity) and their frost resistance (r = 0,86), but the correlation coefficient of this index with frost resistance of plants in tillering phase was significantly lower (r = 0,47). At the same time, a high correlation was found between the content of low-molecular-weight protectors in hardened seedlings and frost resistance of tillering adult plants (r = 0.89). The closest correlation was observed between the integral normalized indicator, comprising the sum of normalized values of antioxidant enzymes activity and the content of low-molecular-weight protectors in hardened seedlings, and frost resistance of seedlings (r = 0,94) and plants in tillering phase (r = 0,89). A presence of specific features in the functioning of antioxidant system during cold adaptation of cereal seedlings was established. Rye is characterized by a high content of low-molecular-weight protective compounds; at the same time, increased activity of antioxidant enzymes - superoxide dismutase and catalase - was noted in wheat seedlings. In triticale, depending on the genotype, the values of both enzymatic antioxidant activity and the content of low-molecular-weight protectors varied.

scholarly journals Root Distribution and Nitrogen Fixation Activity of Tropical Forage Legume American Jointvetch (Aeschynomene americanaL.) cv. Glenn under Waterlogging Conditions

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Manabu Tobisa ◽  
Masataka Shimojo ◽  
Yasuhisa Masuda
Keyword(s):  
Nitrogen Fixation ◽  
Root Distribution ◽  
Water Surface ◽  
Nitrogenase Activity ◽  
Soil Surface ◽  
Nodule Formation ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity ◽  
Net Assimilation ◽  
Waterlogging Treatment

We investigated the root distribution and nitrogen fixation activity of American jointvetch (Aeschynomene americanaL.) cv. Glenn, under waterlogging treatment. The plants were grown in pots under three different treatments: no waterlogging (control), 30 days of waterlogging (experiment 1), and 40 days of waterlogging (experiment 2). The plants were subjected to the treatments on day 14 after germination. Root dry matter (DM) weight distribution of waterlogged plants was shallower than controls after day 20 of waterlogging. Throughout the study period, the total root DM weight in waterlogged plants was similar to that in the controls. Enhanced rooting (adventitious roots) and nodule formation at the stem base were observed in waterlogged plants after day 20 of waterlogging. The average DM weight of individual nodules on the region of the stem between the soil surface and water surface of waterlogged plants was similar to that of individual taproot nodules in the controls. Waterlogged plants had slightly greater plant DM weight than the controls after 40 days of treatment. The total nitrogenase activity (TNA) of nodules and nodule DM weight were higher in waterlogged plants than in the controls. Waterlogged American jointvetch had roots with nodules both around the soil surface and in the area between the soil surface and water surface after 20 days of waterlogging, and they maintained high nitrogenase activity and net assimilation rate that resulted in an increased growth rate.

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