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Section 1. Biology
Ismailova Karomat Mahmudjonovna, Ablakulova Nodira Abduvaxobovna, Kushiev Khabibjon Khojiboboevich, Gulistan State University, Uzbekistan E-mail: [email protected]
Fungitoxic and growth-promoting propoties of the complexs copper component and glycyrrhizin acid
Abstract: It has been defined height stimulation and fungi toxic activity of the complex glycerin acid with a copper component. The complex created under local materials glycerin acid and a copper component. The stability mechanism of rust to disease of wheat on the formation of phenol compounds was studied. The development of the Puccinia striformis f.sp.tritici from concentration of phenol compounds in wheat fabrics is shown dependence. It is defined that the formation of phenol compounds is increased under the influence of complex that it has a positive impact on stability of plants to rust.
Keywords: Wheat, a yellow rust, glycerin acid, copper component, phenol compounds.
For the last years in the region of Central Asia phytopathologic situations have aggravated in connection with epiphithotical distributions of a rust (yellow and brown), sheet spotty (helmintiosporios, physiological and yellow spots) and others which puts considerable economic damages, reducing a crop and quality of grain of wheat.
The yellow rust is developed on wheat, both on irrigation, and on arid lands [1] and is observed frequent flashes with presence of natural sources of an infection [2]. The capacity of penetration of spore is more than wheat growing under irrigation conditions that the vegetative weight is developed, a mouth of leaf is found opened longer. The affection of wheat leads to decrease in assimilation activity, in intensification transpiration of breath and breach of other physiological and biological processes. There is a decrease in starch and protein synthesis in endosperm that reduces the quality and weight of grain [3-5]. It is observed the changes in balance of regulators of growth in fabrics of plants: indolacetic acid (IAA) [6] is accumulated, ethylene is intensively formed, the contents of absizine acid is increased. It results in loss of a crop and its quality.
The protection ofplants is the important condition of the intensification of agriculture. Therefore, together with agrotechnical and selection methods for struggle against fungi diseases, various chemical preparations are widely used just against a yellow rust of wheat.
The application of chemotherapeutical substances has an object to protect plants from an infection not only in the present year, but also in the subsequent generations. Thanks to this resistance to a parasite is increased both in a year of application, and in the subsequent generations. Besides, growth and development of plants is stimulated and in consequence the crop is increased and the quality of production is improved [7].
Lately various chemical preparations have been widely applied in struggle for wheat rust.
Bayleton, baytan and others fungicides of triazole compounds possess some properties of cytokinine in cytokininesensitive tests [6] and also are characterized multiprotector impact on plants as the plants preprocessed by them, are less damaged under the influence ofnot only biotic, but also such abiotic stress factors, as high and low temperatures, a drought [8-12]. It is necessary to emphasis that all these fungicides
are effective against various kinds of fungies. But all of them are imported, therefore it is necessary to conduct work on creation from accessible raw materials of the domestic cheap, ecologically safe preparations, which are capable to enhance resistance ofplants to fungi diseases.
In connection with this we worked out a stimulator (complex glycerin acid and a copper component) for the growth of plants, possessing fungitoxic on the basis of glycyrrhizin acid and copper component (GACC).
The objective of this inquire is to investigate antirust activity of this preparation and find out feasible mechanisms of active resistance of wheat to rusts, connected with phenolic metabolisms.
Materials and methods
We conducted an investigation on plants of winter soft wheat (Triticum avestium) sorts of Sayhun, Bayaut 1, Sanzar 8, Chillaki and Yonbosh in laboratory and field conditions. Before sowing seeds of wheat we processed a solution of GACC. Sowing the samples of wheat on an infectious site pickled by preparation GACC and without it was in 1 m 2 50-60 grains with row-spacing 20 sm. And we have planted special pots by 10 pieces in laboratory conditions pickled and control samples of grains of wheat.
We studied antifungal and growstimulation activity of the GACC preparation. We compared the antifungal activity of GACC with fungicides of bayleton and tilt. For this purpose, we sprayed the solution of GACC and fungicides of bayleton and tilt before and after artificial infections of a yellow rust.
Artificial infection with local populations of a yellow rust of adult plants on an infectious site in field conditions and sowed in special pots in laboratory conditions we spent in the period of vegetation ofwheat. For inoculation plants we mixed spores with talc in the ratio 1:50 and sprayed mixture on plants. After that, we covered affected plants with polyethylene and put the into refrigerator within 5 days. As for field conditions affected plants were isolated from other wheat fields and watered 7-8 times to increase humidity of soil.
We made an accounting of affection in 14-18 days after infection. The degree of affection of samples was estimated on a percentage scale.
The definition of phenol compounds, both free and connected, in various parts of the specified plants was made on work
of Berner [13]. After gathering fresh material was directly divided into separate parts (leaves, an ear or an inflorescence, a stalk and roots). The divided material was frozen, exposed to liofil drying and was stored at temperatures till the analysis in such kind.
We filled 5-10 gr. the dry crushed material with water (1:10) and insisted at 26 °C within 24 hours in toluol presence. And we filtered the received extract from a vegetative material, acidified till pH 3.0-3.5 and repeatedly extracted with ether. We dried up the connected ether extracts over calcinated Na2SO4 and made them evaporate at low temperature dry. The rest after ether removal was dissolved in 1-2 ml. by 96 %-s' ethyl spirits and subjected chromato-graphic to division into a paper.
In order to reveal stains on chromatograms we used solutions of dinitrogenecting sulfonil acids and n-nitroaniline, ethanol solution
120 -t
floroglucyne and concentrated HCI, a solution 2N 2.4-dinitrofe-nilgidrazina.
Results
At epiphithotical developments of a rust and septorios, agro-technical receptions are inefficient and there are huge losses of a crop. In connection with this, constant attention was paid to work out chemical receptions of protection of crops of wheat [14].
We studied growstimulation and fungitoxic properties of the preparation of GACC created by us in the whole period of vegetation of winter grades of wheat. It was observed that marked differences of impact of the preparation of GACC and fungicides of Bayleton and tilt on developments of a yellow rust and biological signs of wheat after inoculation with spores of a rust (Fig. 1 and Table 1).
100 |—|
•mmm
GACC (I) GACC (II) GACC (I) Bayleton Tilt
+ GACC (II) Fungicides □ Bayaut-1 «Saykhun nYonbosh nSanzar-8 ■ Chillaki
Fig. 1. Fungitoxic effects of fungisides of bayleton, tilt, and GACC Table 1. - Influence of preparation GACC and fungicides of bayleton and tilt on biological signs of sorts of wheat
Sorts Scheme of experience Mass 1000 Grain Crop grain, t/ha Supplement
t/ha %
Bayaut-1 Control 40.2 4.75
GACC (I) 44.2 5.32 0.57 12
GACC (II) 43.4 5.03 0.28 5.9
GACC (I) + GACC (II) 44.5 5.76 1.01 21.3
Bayleton 43.6 5.12 0.46 9.7
Tilt 43.3 4.98 0.23 4.8
Saykhun Control 39.3 4.12
GACC (I) 43.1 4.98 0.86 20.9
GACC (II) 44 4.71 0.59 14.3
GACC (I) + GACC (II) 44.7 5.56 1.44 35
Bayleton 44.1 4.66 0.54 13.1
Tilt 43.2 4.45 0.33 8
Sanzar-8 Control 40.1 4.32
GACC (I) 44 5.32 1.0 23.2
GACC (II) 45.5 4.83 0.51 11.8
GACC (I) + GACC (II) 45.7 5.45 1.13 26.2
Bayleton 44.3 4.56 0.24 5.6
Tilt 43.4 4.5 0.18 4.2
Chillaki Control 37.9 3.91
GACC (I) 43 4.43 0.52 13.3
GACC (II) 44.4 4.37 0.46 11.8
GACC (I) + GACC (II) 44.9 4.76 0.85 21.7
Bayleton 42.8 4.32 0.32 8.2
Tilt 43.2 4.23 0.31 7.9
Yonbosh Control 40.4 4.55
GACC (I) 43.5 4.98 0.43 9.5
GACC (II) 44 4.71 0.16 3.5
GACC (I) + GACC (II) 44.6 5.12 0.57 12.5
Bayleton 43.4 4.66 0.11 2.4
Tilt 43.2 4.61 0.06 1.3
During the conducted experiences we observed a positive influence of the preparation of GACC on decrease in affection of sorts of wheat by a yellow rust in comparison with known fungicides of bayleton, tilt. The influence of GACC preparation shows a special attention after spraying. While in the first variant of experience (I) seeds of sorts of wheat is pickled before the sowing, in the second variant (II) preparation was sprayed before inoculation spores of a yellow rust, and in the third variant (I) + (II) seeds of sorts of wheat is pickled with the preparation of GACC before sowing and conducted spraying before inoculation.
The received results show diagnostics and a condition of fields by the period of performance of these researches. Apparently from the data affection of sorts of wheat of Sayhun, Yonbosh are approximately identical, a rather low sensitive Bayaut-1, Sangzar-8 and Chillaki are high affected. GACC is rather effective, which on 100 % protects from a yellow rust of Bayaut-1, 85 % of Sayhun and Yonbosh and on 65 % of Sanzar-8 and Chillaki.
At the same time bayleton shows 83 % of efficiency at sorts of Bayaut-1 and Sayhun, 80 % at Yonbosh, 70 % at Sanzar-8 and 60 % at Chillaki. As for tilt, it protects from desease by yellow rust a sort of Bayaut on 70 %, Sayhun and Yonbosh on 60 %, Sanzar-8 and Chillaki of 50 % accordingly.
Moreover, on the conducted experience on protection of winter sorts of wheat with usage of the preparation of GACC
and fungicides of bayleton and tilt are counted up the productivity of sorts of wheat (Table 1). The received results show that the appreciable supplement of productivity of grains of wheat is observed at two-multiple processing of wheat with the preparation of GACC: before sowing and in the period of vegetation of wheat.
Thus, the created GACC preparation on the basis of natural compound proved much more effective than the fungicides of bayleton and tilt.
It has been defined the presence of phenolic compounds in fabrics of samples ofwheat after inoculation of disputes of a yellow rust (table 2). Phenolic compound, pickling doesn't admit the growth of rust fungis. It enhances the resistence of wheat to desease.
It is important to make comparison between the impact of preparation GACC and fungicides of bayleton and tilt on the formation of wheat phenolic compounds. Therefore we took 100 mg. phenol compound and we put its solution on paper chromatography. And then we isolated separately all phenolic compound. We defined them through the standard phenol compounds. They are following: Ferulic acid (Rf-0.31), n-kumaric acid (Rf-0.46), Cofein acid (Rf-0.28), n-oxybenzoic acid (Rf-0.61), gall acid (Rf-0.72), hallocatehin (r-0.49), catehin (R-0.64), kvercetin (Rf-0.70) kempherol (Rf-0.77), rutine (Rf-0.64). Then we determined the quantity of phenolic compounds (table 2).
Table 2. - Changing in the quantity of phenolic compounds in wheat under the influence of GACC and Bayleton
Scheme of experience Phenol compounds Saykhun Chillaki Sanzar-8
Mass of phenol quantity, mk/100 gr
Control Ferulic acid 155 143 151
n-kumaric acid 132 101 112
Cofein acid 99 112 123
n-oxybenzoic acid 25 33 41
Gall acid 89 77 83
Hallocatexin 91 89 93
Catexin 77 100 106
Bayleton Ferulic acid 156 161 149
n-kumaric acid 138 98 124
Cofein acid 102 117 133
n-oxybenzoic acid 25 36 39
Gall acid 98 85 95
Hallocatexin 108 95 111
Catexin 71 113 99
GACC Ferulic acid 405 342 311
n-kumaric acid 198 168 177
Cofein acid 122 171 189
n-oxybenzoic acid 59 55 52
Gall acid 213 198 208
Hallocatexin 169 137 145
Catexin 131 127 128
It turns out the quantity of ferulic acid was by far high both in control and under the influence of GACC and bayleton. However, n-oxybenzoic acid was made minimal quantity than others. Besides, the table dates showed that a number of phenolic compounds were changed little than the influence of GACC.
The results of the investigation showed that the quantity of ferulic acid, n-kumaric acid, gall acid, hallocatexin, and katex-in is increased under the influence of GACC preparation. It is known [15], these phenolic compounds display high physiological activity on the formation of lignin and protect plants from biotic and abiotic stresses [16-19].
Discussion
GACC was characterized by the property of immunostimulus that has been mainly shown by cytological methods in relation to pathogen causing a rust. In modeling experiences it has been also established that GACC behaves as a regulator of growth possessing cytokinin effect (Dalimov and Kushiev). While in structure GACC there is glycyrrhizin acid.
From the literary data it is known [20-21] that glycyrrhizin acid in concentration of 10-8-10-6 M has stimulating action of plants. Besides, in laboratory tests it is established that glycyrrhizin acid and its aglycon glycyrrhitic acid in concentration
of 10-8-10-6 M increase indicators of growing cotton seeds. The maximum stimulating effect was observed in concentration (10-7). On physiological tests on auccyne glycyrrhizin acid of 10-7-10-6 M similarly IAA stimulates a gain of primary growing of wheat and the formation of roots at sheet shanks of a string bean, and in concentration of 10-8-10-7 M also stimulates S-IAA transport in primary growing of a cotton.
Proceeding from it is possible to tell that GACC has stimulating activity and consequently shows high efficiency in the conducted experiments.
Besides the preparation of GACC contains a biological active copper component in the structure. The role of the copper in a life of plants is rather specific: copper cannot be replaced by any other element or their sum.
Bringing in high doses of nitric fertilizers strengthens requirement of plants for copper and promotes an aggravation of symptoms of copper insufficiency. It specifies that copper plays an important role in a nitric exchange and makes a peculiar part of specific enzymes structure which participates at biosynthesis im-munomodelators.
One of the immunity mechanisms is capacity of plants to form phenol compounds in reply to antibiotic substances infection. As for characteristic features of coppercontained proteins that they enhance of resistance of plants against fungi diseases by stimulation of synthesis of phenol compounds in cells of plants.
Lately it has been obvious that phenol compounds in comparison with simple structure are capable to carry out a role of warning (signal) substances in interrelations of a plant — owner with fungus and some parasitizing plants. The Plant — owner synthesises these signal substances, parasitizing organisms respond to their the genes, which are necessary for realization of the subsequent stages of interaction.
From conducted experimental researches we have determined that many phenolic compounds considerably suppress the vital activity of fungies, including rusts ofwheat. Besides there are many experimental data showing higher toxicity of oxidised polyphenols in comparison with their restored forms. From here there is an assumption that during the affection of plants an oxidation polyphenols of oxidising enzymes are arised (in particular, polyphenoloxidaz) with formation of toxic substances which it is more active than the restored polyphenols, oppress the growth of fungi. As to the formation of phenoloxydaz, it depends on an available microelement of copper. Thus, from an available copper component in the structure of GACC the formation of phenoloxydaz is entensified that it will lead to the oppression of fungi with the increase in the quantity of phenolic compounds (Table 2).
Thus, glycyrrhizin acid available in the structure of GACC stimulates the growth and development of wheat, and the copper component suppresses the development fungus with stimulation the formation of phenol compounds.
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