No disruption of rhizobial symbiosis during early stages of cowpea domestication

Annexins as Ca2+/phospholipid-binding proteins are involved in the control of many biological processes essential for plant growth and development. In a previous study, we had shown, using a proteomic approach, that the synthesis of two annexins is induced in pea roots in response to rhizobial inoculation. In this study, phylogenetic analysis identified these annexins as PsAnn4 and PsAnn8 based on their homology with annexins from other legumes. The modeling approach allowed us to estimate the structural features of these annexins that might influence their functional activity. To verify the functions of these annexins, we performed comparative proteomic analysis, experiments with calcium influx inhibitors, and localization of labeled proteins. Essential down-regulation of PsAnn4 synthesis in a non-nodulating pea mutant P56 (sym10) suggests an involvement of this annexin in the rhizobial symbiosis. Quantitative RT-PCR analysis showed that PsAnn4 was upregulated at the early stages of symbiosis development, starting from 1–3 days after inoculation to up to 5 days after inoculation, while experiments with the Ca2+ channel blocker LaCl3 revealed its negative influence on this expression. To follow the PsAnn4 protein localization in plant cells, it was fused to the fluorophores such as red fluorescent protein (RFP) and yellow fluorescent protein (YFP) and expressed under the transcriptional regulation of the 35S promoter in Nicotiana benthamiana leaves by infiltration with Agrobacterium tumefaciens. The localization of PsAnn4 in the cell wall or plasma membrane of plant cells may indicate its participation in membrane modification or ion transport. Our results suggest that PsAnn4 may play an important role during the early stages of pea-rhizobial symbiosis development.

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Realization of protective and symbiotic properties of soybeans using fungicide seed treatment

Agricultural science and practice ◽

10.15407/agrisp8.02.024 ◽

2021 ◽

Vol 8 (2) ◽

pp. 24-35

Author(s):

T. Mamenko ◽

S. Kots ◽

V. Patyka

Keyword(s):

Lipid Peroxidation ◽

Ascorbate Peroxidase ◽

Seed Treatment ◽

Root Nodules ◽

Nodule Bacteria ◽

Early Stages ◽

Nitrogen Fixation Activity ◽

Seed Inoculation ◽

Rhizobial Symbiosis ◽

Peroxidation Processes

Aim. The elaboration of efficient legume-rhizobial symbiosis systems, involving active strains of nodule bacteria, in the combination with fungicide seed treatment may be an alternative method of providing ecologically friendly nitrogen sources to plants and promoting their tolerance to the external factors, which is relevant for preservation and restoration of envi- ronmental quality. Therefore, the aim of our study was to determine the impact of pre-sowing seed treatment with fungi- cides, which differ in the action spectrum of active substances – Standak Top (fipronil, 250 g/l, thiophanate-methyl, 225 g/l, pyraclostrobin, 25 g/l) and Maxim XL (fludioxonyl, 25 g/l, metalaxyl, 10 g/l), on the intensity of the development of lipid peroxidation processes, the activity of antioxidant enzyme ascorbate peroxidase and nitrogen fixation activity in soybeans on the early stages of forming legume-rhizobial symbiosis. Methods. Microbiological (cultivation of a bacterial culture, seed inoculation), physiological (vegetative experiment), biochemical (spectrophotometric determination of the content of lipid peroxidation products and the activity of ascorbate peroxidase; measuring the nitrogen-fixation activity using a gas chro- matography). Results. It was found that pre-sowing fungicide treatment of soybean seeds and subsequent inoculation with active rhizobia of strain 634b did not result in the change in the content of TBA-active products in roots and root nodules (the values of indices were within the experiment deviation). At the same time, after seed inoculation using rhizobia and treat- ment with Maxim XL, there was an increase in the activity of ascorbate peroxidase in the roots from 20.3 to 30.8 %, and with Standak Top – from 20.0 to 29.8 % during the early stages of ontogenesis till the formation of the third ternate leaf. Here, the activity of the enzyme in root nodules increased by 24.7–40.3 % at the fungicidal effect. Our data demonstrate that the combination of fungicide seed treatment and inoculation with active rhizobia does not induce lipid peroxidation processes, but promotes the initiation of protective antioxidant properties in soybeans. It is accompanied with efficient functioning of the symbiotic apparatus, which is manifested in the increase in nitrogen-fixing activity of nodule bacteria, formed by active rhizobia of strain 634b after the seed treatment with Standak Top – by 98.3 and 78.1 % and after Maxim XL – by 78.6 and 196.2 % respectively, during the stages of the second and third ternate leaves. Conclusions. The pre-sowing soybean seed treatment with fungicides Standak Top and Maxim XL and the subsequent inoculation with active rhizobia of strain 634b does not induce the development of lipid peroxidation processes, but increases the activity of the antioxidant enzyme, ascor- bate peroxidase, in the roots and root nodules, which is accompanied with the efficient work of the symbiotic apparatus on the early stages of determining legume-rhizobial symbiosis. This method of seed treatment may be a novel measure, to use in the technologies of cultivating soybeans to enhance the realization of the symbiotic potential and meet the needs of plants in ecologically friendly nitrogen, and to promote the formation of their tolerance to the corresponding cultivation conditions.

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Cytokinin Content during Early Stages of Legume-Rhizobial symbiosis and effect of hypothermia

Russian Journal of Plant Physiology ◽

10.1134/s1021443712030028 ◽

2012 ◽

Vol 59 (5) ◽

pp. 656-661 ◽

Cited By ~ 5

Author(s):

G. P. Akimova ◽

M. G. Sokolova

Keyword(s):

Cytokinin Content ◽

Early Stages ◽

Rhizobial Symbiosis

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The intensity of ethylene release by soybean plants under the influence of fungicides in the early stages of legume-rhizobial symbiosis

Regulatory Mechanisms in Biosystems ◽

10.15421/022014 ◽

2020 ◽

Vol 11 (1) ◽

pp. 98-104

Author(s):

T. P. Mamenko ◽

S. Y. Kots ◽

Y. O. Khomenko

Keyword(s):

Nitrogen Fixation ◽

Virulent Strain ◽

Root Nodules ◽

Symbiotic System ◽

Early Stages ◽

True Leaf ◽

Rhizobial Symbiosis ◽

Soybean Plants ◽

Phytohormone Synthesis ◽

Highly Virulent

The effect of pre-sowing treatment of soybean seeds with fungicides on the intensity of ethylene release, the processes of nodulation and nitrogen fixation in different symbiotic systems in the early stages of ontogenesis were investigated. The objects of the study were selected symbiotic systems formed with the participation of soybean (Glycine max (L.) Merr.) Diamond variety, strains Bradyrhizobium japonicum 634b (active, virulent) and 604k (inactive, highly virulent) and fungicides Maxim XL 035 PS (fludioxonil, 25 g/L, metalaxyl, 10 g/L), and Standak Top (fipronil, 250 g/L, thiophanate methyl, 225 g/L, piraclostrobin, 25 g/L). Before sowing, the seeds of soybean were treated with solutions of fungicides, calculated on the basis of one rate of expenditure of the active substance of each preparation indicated by the producer per ton of seed. One part of the seeds treated with fungicides was inoculated with rhizobium culture for 1 h (the titre of bacteria was 107 cells/mL). To conduct the research we used microbiological, physiological, biochemical methods, gas chromatography and spectrophotometry. It is found that, regardless of the effectiveness of soybean rhizobial symbiosis, the highest level of ethylene release by plants was observed in the stages of primordial leaf and first true leaf. This is due to the initial processes of nodulation – the laying of nodule primordia and the active formation of nodules on the roots of soybeans. The results show that with the participation of fungicides in different symbiotic systems, there are characteristic changes in phytohormone synthesis in the primordial leaf stage, when the nodule primordia are planted on the root system of plants. In particular, in the ineffective symbiotic system, the intensity of phytohormone release decreases, while in the effective symbiotic system it increases. At the same time, a decrease in the number of nodules on soybean roots inoculated with an inactive highly virulent rhizobia 604k strain due to the action of fungicides and an increase in their number in variants with co-treatment of fungicides and active virulent strain 634b into the stage of the second true leaf were revealed. It was shown that despite a decrease in the mass of root nodules, there is an increase in their nitrogen-fixing activity in an effective symbiotic system with the participation of fungicides in the stage of the second true leaf. The highest intensity of ethylene release in both symbiotic systems was recorded in the stage of the first true leaf, which decreased in the stage of the second true leaf and was independent of the nature of the action of the active substances of fungicides. The obtained data prove that the action of fungicides changes the synthesis of ethylene by soybean plants, as well as the processes of nodulation and nitrogen fixation, which depend on the efficiency of the formed soybean-rhizobial systems and their ability to realize their symbiotic potential under appropriate growing conditions.

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Spherulitic crystal growth in the prodissoconch larval of Crassostrea virginica

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100076263 ◽

1983 ◽

Vol 41 ◽

pp. 518-519

Author(s):

George G. Cocks ◽

Louis Leibovitz ◽

DoSuk D. Lee

Keyword(s):

Crystal Structure ◽

Crassostrea Virginica ◽

Crystal Orientation ◽

Developmental Changes ◽

Gastrula Stage ◽

Orthorhombic Crystal ◽

Orthorhombic Crystal Structure ◽

Stages Of Growth ◽

Early Stages ◽

Initial Deposition

Our understanding of the structure and the formation of inorganic minerals in the bivalve shells has been considerably advanced by the use of electron microscope. However, very little is known about the ultrastructure of valves in the larval stage of the oysters. The present study examines the developmental changes which occur between the time of conception to the early stages of Dissoconch in the Crassostrea virginica(Gmelin), focusing on the initial deposition of inorganic crystals by the oysters.The spawning was induced by elevating the temperature of the seawater where the adult oysters were conditioned. The eggs and sperm were collected separately, then immediately mixed for the fertilizations to occur. Fertilized animals were kept in the incubator where various stages of development were stopped and observed. The detailed analysis of the early stages of growth showed that CaCO3 crystals(aragonite), with orthorhombic crystal structure, are deposited as early as gastrula stage(Figuresla-b). The next stage in development, the prodissoconch, revealed that the crystal orientation is in the form of spherulites.

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Formation of Dislocation Channels in Neutron Irradiated Molybdenum

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100096618 ◽

1972 ◽

Vol 30 ◽

pp. 672-673

Author(s):

S. Mahajan

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Ambient Temperature ◽

Room Temperature ◽

Channel Formation ◽

Early Stages ◽

Transmission Electron ◽

Three Stages ◽

Two Stages ◽

Polycrystalline Molybdenum

The evolution of dislocation channels in irradiated metals during deformation can be envisaged to occur in three stages: (i) formation of embryonic cluster free regions, (ii) growth of these regions into microscopically observable channels and (iii) termination of their growth due to the accumulation of dislocation damage. The first two stages are particularly intriguing, and we have attempted to follow the early stages of channel formation in polycrystalline molybdenum, irradiated to 5×1019 n. cm−2 (E > 1 Mev) at the reactor ambient temperature (∼ 60°C), using transmission electron microscopy. The irradiated samples were strained, at room temperature, up to the macroscopic yield point.Figure 1 illustrates the early stages of channel formation. The observations suggest that the cluster free regions, such as A, B and C, form in isolated packets, which could subsequently link-up to evolve a channel.

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