Influence of physiologically active substances on the anatomical structure of mung bean sprouts when using potassium humate Agro.Bio

In recent years, literature has widely noted the effective action of physiologically active substances on the anatomical structure of plants. The positive effect of growth-accelerating substances lies in their participation in regulating protein synthesis, influencing labile forms of nucleic acids and primarily mRNA.

Growth and morphogenesis processes are strictly coupled with changes in nucleic acid content, their localization in the cell, and their state in the cytoplasm and nucleus. The formation of cellular structures is preceded by the accumulation of nucleic acids in these locations. Since physiologically active substances enhance nucleic acid metabolism, they should accelerate the formative processes of the entire plant organism.

There is also data on the influence of differentiated inhibitors on plant growth and development. The action of 8-azaguanine - a guanine analog - manifests in its incorporation into labile RNA forms, distorting the matrix, resulting in the production of proteins with altered structure. Since formative processes in the cell are closely related to protein metabolism, it can be assumed that the effect of azaguanine must necessarily manifest in the anatomical structure of plants.

Another inhibitor - 2,4-dinitrophenol, while significantly activating respiration, disrupts oxidative phosphorylation, which affects the functional activity of the entire organism and cannot but influence its anatomical structure.

Physiologically active substances increase the energy potential of the plant, form adenosine triphosphate (ATP), and thereby promote the regeneration of nucleoside diphosphates into nucleoside triphosphates, leading to the activation of nucleic acid synthesis. Accordingly, 8-azaguanine, 2,4-dinitrophenol, ATP, and potassium humate were included in the experimental scheme.

Objective:

studying the influence of these substances on the anatomical structure of mung bean sprouts and the removal of inhibitor effects by growth-stimulating substances.

Research methodology

Seeds were soaked in water, ATP (1.4·10⁻⁵ M), potassium humate (3.1·10⁻⁵ M), 8-azaguanine (10⁻⁴ M), and 2,4-dinitrophenol (10⁻³ M). After 24 hours, sprouts with inhibitors were transplanted to water, ATP, and potassium humate, allowing observation of the influence of all listed substances on sprout growth and development, as well as the effect of removing inhibitory action.

Anatomical studies were conducted on eight-day-old mung bean sprouts. The leaf, stem, and root were examined. Temporary leaf preparations were made to determine the number of stomata and the size of guard cells.

The main part of the research was performed on permanent preparations fixed according to Carnoy and Navashin. The work technique (from sampling to obtaining preparations) corresponded to generally accepted methodologies.

Sections of paraffin-embedded material were prepared using a sliding microtome. Section thickness varied from 8 to 20 μm depending on cell size.

Quantitative-anatomical studies of the stem and root included:

• Determination of the overall section diameter and area.
• Ratio of tissues in cross-sections.

Average values were calculated based on 10 measurements and examination of 10 sections per variant (100 measurements total). Root sections were made at its base, stem sections in the lower part of the hypocotyl.

Section staining was performed with dyes:

• Hematoxylin with eosin according to Heidenhain.
• Methyl green with pyronin according to Unna and Brachet.

Research results

Differences in the anatomical structure of mung bean sprouts grown on different backgrounds concern quantitative ratios, the nature of development, and the degree of tissue differentiation in the stem, root, and leaf.

Inhibitory effects:

• 8-azaguanine and 2,4-dinitrophenol cause disruption of intracellular metabolism, death of integumentary tissues, cell deformation, inhibition of seed germination, and sprout growth.

Stimulating effects:

• ATP and potassium humate increase the overall thickness and area of stem and root tissues.
• When transplanting sprouts from inhibitors to ATP or potassium humate, growth recovery and tissue formation are observed.

Table 1. Influence of substances on the tissue ratio in mung bean sprout stems

Seed soaking medium	Transplant medium	Indicators
		Average section diameter, μm	Section area, mm²	Tissue thickness by radius, μm	Tissue area, mm²	Total section area, % of control	Bundle and parenchyma zone, % of control
Water	Water (control)	127.4	0.012	25.8 / 37.9	0.008 / 0.001	100	100
ATP, 1.4·10⁻⁵	ATP, 1.4·10⁻⁵	187.2	0.026	60.1 / 33.5	0.023 / 0.003	216	287
Potassium humate, 3.1·10⁻⁵	Potassium humate, 3.1·10⁻⁵	168.8	0.022	52.4 / 32.4	0.020 / 0.002	183	250
8-azaguanine, 10⁻⁴	8-azaguanine, 10⁻⁴	72.0	0.004	16.5 / 24.5	0.0036 / 0.0004	33	45
8-azaguanine, 10⁻⁴	Water	99.2	0.006	23.8 / 25.7	0.004 / 0.002	50	50
8-azaguanine, 10⁻⁴	ATP, 1.4·10⁻⁵	175.0	0.024	16.1 / 41.4	0.014 / 0.010	200	175
8-azaguanine, 10⁻⁴	Potassium humate, 3.1·10⁻⁵	140.4	0.015	43.2 / 27.0	0.010 / 0.005	125	125
2,4-dinitrophenol, 10⁻³	2,4-dinitrophenol, 10⁻³	100.9	0.007	20.2 / 30.2	0.006 / 0.001	58	75
2,4-dinitrophenol, 10⁻³	Water	109.4	0.009	27.7 / 27.0	0.007 / 0.002	75	87
2,4-dinitrophenol, 10⁻³	ATP, 1.4·10⁻⁵	172.8	0.022	51.8 / 34.5	0.018 / 0.004	183	225
2,4-dinitrophenol, 10⁻³	Potassium humate, 3.1·10⁻⁵	159.8	0.019	48.2 / 31.6	0.016 / 0.003	158	200

Table 2. Influence of substances on the tissue ratio in mung bean sprout roots

Seed soaking medium	Transplant medium	Indicators
		Average section diameter, μm	Section area, mm²	Tissue thickness by radius, μm	Tissue area, mm²	Total section area, % of control	Phloem-xylem zone, % of control
Water	Water (control)	282.5	0.25	46.7 / 94.5	0.17 / 0.08	100	100
ATP, 1.4·10⁻⁵	ATP, 1.4·10⁻⁵	581.4	0.76	179.6 / 111.1	0.65 / 0.11	304	382
Potassium humate, 3.1·10⁻⁵	Potassium humate, 3.1·10⁻⁵	439.6	0.61	52.0 / 90.0	0.28 / 0.13	244	282
8-azaguanine, 10⁻⁴	8-azaguanine, 10⁻⁴	201.7	0.13	No differentiation	---	52	---
8-azaguanine, 10⁻⁴	Water	280.6	0.25	21.5 / 18.8	0.10 / 0.15	100	60
8-azaguanine, 10⁻⁴	ATP, 1.4·10⁻⁵	380.6	0.45	100.0 / 90.0	0.30 / 0.15	180	177
8-azaguanine, 10⁻⁴	Potassium humate, 3.1·10⁻⁵	360.3	0.41	90.0 / 90.1	0.28 / 0.13	164	164
2,4-dinitrophenol, 10⁻³	2,4-dinitrophenol, 10⁻³	267.3	0.22	39.3 / 94.3	0.10 / 0.12	88	59
2,4-dinitrophenol, 10⁻³	Water	280.3	0.24	42.0 / 98.1	0.18 / 0.06	96	105
2,4-dinitrophenol, 10⁻³	ATP, 1.4·10⁻⁵	390.5	0.48	105.0 / 90.2	0.24 / 0.24	192	141
2,4-dinitrophenol, 10⁻³	Potassium humate, 3.1·10⁻⁵	389.3	0.47	104.3 / 90.3	0.22 / 0.25	188	129

Table 3. Influence of substances on the number of stomata and size of guard cells

Seed soaking medium	Transplant medium	Number of stomata (count)	Size of guard cells (μm)
Water	Water (control)	93.2	25.8
ATP, 1.4·10⁻⁵	ATP, 1.4·10⁻⁵	164.4	60.1
Potassium humate, 3.1·10⁻⁵	Potassium humate, 3.1·10⁻⁵	127.0	52.4
8-azaguanine, 10⁻⁴	8-azaguanine, 10⁻⁴	77.6	16.5
8-azaguanine, 10⁻⁴	Water	87.6	23.8
8-azaguanine, 10⁻⁴	ATP, 1.4·10⁻⁵	140.8	51.8
8-azaguanine, 10⁻⁴	Potassium humate, 3.1·10⁻⁵	120.0	48.2
2,4-dinitrophenol, 10⁻³	2,4-dinitrophenol, 10⁻³	71.2	20.2
2,4-dinitrophenol, 10⁻³	Water	81.2	27.7
2,4-dinitrophenol, 10⁻³	ATP, 1.4·10⁻⁵	105.2	51.8
2,4-dinitrophenol, 10⁻³	Potassium humate, 3.1·10⁻⁵	102.8	48.2

Conclusions

1. Adenosine triphosphate and potassium humate Agro.Bio stimulate growth and cell differentiation, which is reflected in the anatomical structure of the root, stem, and leaf of mung bean sprouts.
2. 8-azaguanine suppresses protein metabolism by distorting RNA structure, negatively affecting cell growth and reproduction.
3. 2,4-dinitrophenol inhibits vital cellular processes by disrupting respiration and phosphorylation.
4. ATP and potassium humate partially remove the inhibitory effects of 8-azaguanine and 2,4-dinitrophenol, restoring energy potential and normalizing cell growth.