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Khusanov Tоkhir, Junior researcher, of the Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan E-mail: tohir_husanov@mail.ru Davranov Qodirjan, doctor, of Biological Sciences, National university of Uzbekistan after named Mirzo Ulugbek Vakhabov Abdurasul, doctor, of Biological Sciences, professor, National university of Uzbekistan after named Mirzo Ulugbek
EFFECT OF ALFALFA MOSAIC VIRUS ON THE CONTENT OF MICRO AND MACROELEMENTS IN ALFALFA PLANTS IN UZBEKISTAN
Abstract: Leaf samples showing mosaic virus symptoms were collected for analyses from alfalfa growing areas in botanical garden of The National university of Uzbekistan. Alfalfa mosaic virus (AMV) was detected in the plant samples by using mechanical inoculation of test plants and visual determination. The macro and microelements were assayed in the infected plant samples in order to determine the effects of AMV on the nutrient content of alfalfa. The content of micro and macro elements decreased, as well as the amount of peroxidase in the infected alfalfa leaves as compared to the healthy plant samples.
Keywords: alfalfa mosaic virus, peroxidase, micro and macro-elements, photosynthesis.
Introduction an important role in the plant cell [2, 4]. Cu+2 is an ele-
Microelements as cobalt, boron, manganese, zinc, ment affecting the amount of chlorophyll, decrease in
molybdenum, copper, iron, which are required for the the amount of this microelement adversely affects the
normal growth and development of plants, directly par- process of photosynthesis. Lack of iron element leads to
ticipate in metabolism. Also they play important role in chlorosis of the plant [1].
the management of enzyme systems involved in com- Based on the above stated, this article describes re-
plex biochemical and physiological process occurring search on the effect of viral damage to lupine plants, on
in body. Trace elements are important in the life pro- the change in the number of micro and macro-elements
cesses of plants, animals and people, if their number is and the activity of the enzyme peroxidase. not enough or more, or even none, then metabolism is Materials and Methods
disrupted [1, 2, 3, 4]. Long-term studies conducted by Samples exhibiting mosaic virus symptoms were col-
physiologists and agro chemists have shown that in order lected from growing areas in botanical garden of National
to fully grow and develop such basic crops as cotton, al- university of Uzbekistan. We weighed the initial mass of
falfa, corn, etc., microelements should be in the appropri- samples, then dried them in an oven to absolute weight
ate amount in the soil [1, 3, 5]. In plants, the cations Na+ and re-weighed and stored in paper bags. For the study,
and Cu+2, passing through semipermeable membranes of slurry of a certain number of samples was ground to a
the cell wall, are considered to be elements that change homogeneous mass, then the amount of microelements
the pH of the cell sap. Therefore, these elements play was analyzed by neutron activation [6, 7, 8].
To determine the activity of the enzyme peroxidase in the infected plants, samples from alfalfa leaves of weakly, medium and heavily infected and healthy plants of 10 ml were weighed on an electronic scales in the ratio 1: 1 (10 ml) with the addition of acetate (CH3COOH) buffer (pH- 4.7 0.04 M), ground in a porcelain mortar to a uniform mass, filtered through 4 layers of gauze, cen-trifuged for 15 minutes at 4000 rpm to precipitate the constituent parts of the cell. The supernatant was mixed in numbered tubes in the presence of the enzyme peroxidase, determined by the Boyarkin method. In order to achieve this, 2 drops of supernatant, 0.4 ml of benzidine
A
solution, 0.2 ml of distilled water and 0.2 ml of H2O2 and a photocolorimeter with a red light filter at 640 nm of the wavelength, the activity of the enzyme was determined on the basis of the rate of the reaction [9, 10].
Results and discussion
When monitoring the fields in which alfalfa was grown, its AMV affection was established. A wide spread of the disease in phenological observations was manifested by splitting plant pigments, the symptoms of a mosaic (Fig. 1). It is known that when the virus is severely affected by the virus, the amount of chlorophyll is greatly reduced in plants, and this affects the yield of alfalfa.
B
C
Figure 1. A) strongly injured; B) medium affected; C) healthy plants
The infection with virus leads to a decrease in the pigment in the leaves, a change in the process of photosynthesis and transpiration. In this case, the photochemical activity, transcription and other important physiological processes are violated, which leads to a tangible loss of
the quality of the plant product and the amount of alfalfa biomass.
Alfalfa along with other crops is important in animal husbandry and other branches of the national economy. Because ofthe large accumulation ofnitrogen in the roots
of alfalfa plants, the nutritional value of fodder is much greater. Therefore, it is important to animal husbandry. The literature indicates that the amount of alfalfa affected in the plant is 35%. Alfalfa from the atmosphere gets free nitrogen and transforms it into a state assimilated by the plant. In plants, alfalfa also contains a large number of vitamins, proteins, mineral elements and their residues.
As stated earlier, microelements (boron, manganese, molybdenum, iron, zinc, copper) in minimum quantity are necessary for the normal development of the plant. They participate in the process of metabolism, have a definite place in the management of the action of enzyme complexes involved in complicated processes of organism.
Investigations on the effect of infected alfalfa AMV plants on the composition of micro and macro-elements
Table 1. - Quantity of micro
are given in (Table 1). The data show that when AMV is infected in plant leaves, the amount of Fe, Cu, Zn, Ba, Br, Na and Ca is significantly reduced. Reducing the amount of chlorophyll in a plant cell can lead to physiological changes. Amount of Mg and Cs is increased, the amount of As is not sensitive. The increase or decrease of micro and macroelements in case of infection with a viral disease leads to profound changes in the metabolism of plants, i.e. activity of photosynthesis, transpiration and other important processes.
The cations Na +, Ca+2 in plants passing through semi-prescribed membranes of the cell wall change the ionic strength (pH) of the cell sap.
Reduction of these elements adversely affects the complete course of physiological processes in the plant cell.
-macro-elements in mg
№ Elements Healthy plant, mg Slightly infected, mg Medium infected, mg Strongly infected, mg
1 2 3 4 5 6
1. Mg 3000 3500 3300 4700
2. Cl 965 4100 2850 3500
3. Mn 67 40 43 40
4. Cu 19 13 15 17
5. Na 160 150 99 88
6. R 28000 41000 32000 34000
7. Sm 0.0078 0.012 0.0079 0.0096
8. Mo 13 <0.1 5.0 5.5
9. Lu <0.001 <0.001 <0.001 <0.001
10. U <0.1 <0.1 <0.1 <0.1
11. Au 0.0099 0.0054 0.0092 0.019
12. As <0.1 <0.1 <0.1 <0.1
13. Br 1.1 2.0 3.9 4.7
14. Ca 23000 15100 16100 15700
15. La 0.059 0.054 0.066 0.088
16. Ce 0.13 0.18 <0.1 0.22
17. Hg 0.015 <0.01 0.015 <0.01
18. Th 0.019 0.022 0.034 0.031
19. Cr 0.23 0.36 0.28 0.38
20. Hf 0.014 0.0087 0.0087 0.011
21. Ba 36 21 17 18
22. Sr 230 260 180 170
23. Cs 0.020 0.025 0.042 0.060
24. Ni 4.9 <1.0 8.2 7.1
25. Sc 0.018 0.020 0.023 0.028
1 2 3 4 5 6
26. Rb 8.1 13 14 30
27. Zn 36 40 49 51
28. Co 0.31 0.26 0.40 0.39
29. Fe 110 130 150 130
30. Eu <0.001 0.0026 0.0039 0.0024
31. Sb 0.022 0.027 0.031 0.041
32. Ag <0.01 <0.01 <0.01 <0.01
33. Ti <10 <10 <10 <10
34. V 3.0 <1.0 <1.0 <1.0
35. W <0.1 <0.1 <0.1 <0.1
Cu+2 is a microelement that affects the amount of chlorophyll and a decrease in its amount adversely affects the process of photosynthesis.
In the conducted studies it was found that infecting plants with AMV, the amount of heavy metals in the plant is slightly reduced, and a decrease in the amount of light metals can lead to profound changes in the cell.
The enzyme complex, photosynthesis, respiration and oxidation-regeneration processes are involved in the performance of very important physiological func-
peroxidase enzyme contained in the oxidoreductase is considered a stress enzyme, penetration into the cell of a foreign antigen increases its activity [10].
The peroxidase enzyme is an iron-porphyrin enzyme, in the cell it has a functional dependence in plants, horseradish plants are studied in detail, where the activated oxidative phosphorylation catalyzes the oxidation of pyridine coenzymes. Table 2 shows the results of studies on the enzyme activity in the cell of alfalfa plants when infected with viruses.
tions occurring in plants. According to literary sources,
Table 2.- Influence of virus infection on the activity of the peroxidase enzyme in the leaves of the alfalfa plant (mg / U)
№ Samples Number of replicates Average
Experience 1 Experience 2 Experience 3
1. Healthy alfalfa 0.192 ± 0.03 0.152 ± 0.01 0.143 ± 0.019 0.162 ± 0.01
2. Slightly infected 0.192 ± 0.02 0.166 ± 0.01 0.142 ± 0.02 0.165 ± 0.01
3. Medium infected 0.476 ± 0.06 0.408 ± 0.008 0.363 ± 0.053 0.416 ± 0.008
4. Strongly infected 2.00 ± 0.34 1.428 ± 0.22 1.538 ± 0.11 1.655 ± 0.117
From the results presented, it can be seen that the amount of enzyme and ego activity in healthy and slightly infected plants does not significantly differ. The amount of enzyme and its activity sharply increases in medium and strongly infected plants. When a pathogenic virus enters the cell of the plant, genes are activated that perform stressful functions in extreme cases and, so an increase in the amount and activity of the enzyme may occur. Some literature cited information that this enzyme is a stress enzyme and when affected by various pathogens, and in other extreme cases, its amount increases several times compared to control [9, 10].
Physiological and biochemical processes, including respiration, photosynthesis, transpiration, photo-
chemical reactions, mineral nutrition and many other processes taking place in the living organism occur with the participation of enzymes. In the classification and nomenclature this enzyme is introduced into the group of oxidoreductases, it activates oxidative phosphorylation, catalyzes the oxidation of pyridine coenzymes in a plant cell. Participates in the management of a number of processes associated with oxidation-reduction reactions [10].
The obtained data indicates the expediency of carrying out further experimental studies to clarify the participation of the enzyme in photosynthesis disturbance, transpiration intensity, photochemical activity and a number of other physiological and biochemical
processes following the penetration of the virus into micro and macroelements, pigment complexes, enzyme
the plant cell, and subsequent changes in the number of peroxidase.
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