Physiological and Biochemical Grounding of Different Nanomaterials Use WhenGrowing Corn Seeds

We have investigated in labandfield conditions the influence of cornpre-plant treatment with cuprum and cobalt nano particles and humicacids. We have determined the vitaland morpho-physiological parameters of plants and shown that the effective concentrations are within the limit 0.1-1.0 gper hectare norm of seeding rate. We have discovered that nano particlesi ncrease photosynthesis efficiency andactivate oxidation-reduction ferments and parameters of mineral metabolismthat increases fat in grown seeds by 0.63% as compared with the control.


Introduction
Growth stimulating bio-drugs are important in determining the level of agricultural crops development and their chemical composition.The long-term investigations of the Center of Nanotechnologies and Nanomaterials for the RF Agro-Industrial Complex (FSBEI HPE RSATU) have shown that bio-drugs based on different nanomaterials can be alternative to modern drugs and activate biochemical and physiological processes in plants to increase the agricultural crops yield and quality (Kutskir, Nazarova, Polishchuk, 2012, pp. 135-152;Nazarova, Polishchuk, Stepanova, Churilov, 2014;Golubeva, Ivanycheva, Nazarova, Polishchuk, Churilov, 2013;Polishchuk, Nazarova, Zheglova, 2013, pp. 9-12).
The aim of the investigations is determining in lab conditions the optimal concentrations of biologically active cobalt and cuprum nanoparticles and highly dispersed humic acidswhen treating corn seeds and studying the influence of these nanostructures optimal concentrations on growth, development, productivity and chemical composition of corn seeds and plants.

Method
We had our lab investigations at the Center of Nanotechnologies and Nanomaterials for RF Agro-Industrial Complex at FSBEI HPE RSATU.We used the seeds of the first generation corn hybrid "Obskiy 140".We determined germination energy and lab germination according to GOST 12038-84 "Crops seeds.Methods of determining germination and germination energy" and methods "Determining nanomaterials effect on food and fodder plants in vital and morpho-physiological parameters"developed and tested while working at grant of FCP"Development of RF Nanoindustry Infrastructure for 2008-2011" (Golubeva, Ivanycheva, Nazarova, Polishchuk, Churilov, 2013).We determined germination energy on the third day and lab germination, length and mass of sprouts and roots on the seventh day of the experiment with corn seeds germination.
We had field tests in 2010-2013 at the agrotechnological station of FSBEI HPE RSATU in township Stenkino, Ryazan district of Ryazan oblastand at the demonstration ground of the Ministry of Agriculture at JSC "Agrotechnology" in township Malinishchi, Pronsk district of Ryazan oblast.
We had the experiment according to field tests methods by Dospekhov (Dospekhov, 1985).The plots' arrangement was systematic and the replication number was four.The plots' area was 56 m 2 , the harvesting area was 38 m 2 and replication was evenly even.It was a one factor experiment.The factor was the pre-plant treatment of corn seeds with different biologically active nanosubstances: cobalt and cuprum nanoparticles and ultrafine humic acids.During the vegetation we determined the following parameters: field germination, leaf-area duration, height and mass of plants in vegetation phases, ferments activity, yield and chemical composition of corn seeds.Table 1 presents the scheme of our tests.Ultra-fine humic acids (HA) 0.5; 1.0; 5.0

Results
We have had preliminary lab tests to determine biological activity of cobalt and cuprum nanoparticles.The results have shown that nanoparticles at concentrations of 0.1-10.0g per hectare seeding ratestimulate sprouts development.There is a supposition that nanoparticles have the mode of action different from that of microelements.So as compared with cuprum and cobalt nanoparticles we have investigated the same concentrations of humic acids (soil humus component).We have used humic acids being natural growth stimulatorsin an ultra-fine state.Table 2 presents the facts about germination energy and corn seeds germinationinfluenced by biologically active nanomaterials.The use of cobalt and cuprum nanoparticles and ultra-fine humic acids in lab tests has led to the increase of corn seeds germination energy.Cobalt and cuprum nanoparticles in all concentrations have had the positive result andthe germination energy on the third day of the experiment has on the average been 4.0 % higher than that of the control.We have observed the maximum 3.4 % increase of germination energy as compared with the control under the influence of humic acidsonly at 0.1 g concentration of the active agent.Biologically active nanoparticles of cobalt and cuprum and ultra-fine humic acids have also increased seeds germination.When using cobalt nanoparticles and humic acids the lab germination has exceeded the control one on the average by 1.5% -2%.This parameter value has been 100 % when using 0.1 g cuprum nanoparticles and that proves the high stimulation of growth processes.Table 3 presentsthe facts about the corn sprouts and roots length when treated with nanomaterials on the 7 th day of the experiment.The length of the corn plantlet's aerial portion when treated with cobalt nanoparticles and humic acids has exceeded the control maximum by 9.1% -7.1% correspondingly at concentration of 0.5 g.For cuprum nanoparticles the maximum result has been at active agent concentration of 0.1 g.The positive difference has been 10% as compared with the control.The biologically active substances have improved the plantlet's underground part growth as well.So the roots' length in a case with 0.5 g cobalt nanoparticles treatment has exceeded the control one by 3% and in a case with cuprum nanoparticles by 8%.Ultra-fine humic acids have not changed the root length.
We have also determined the corn sprouts and roots massin the lab tests.It deserves to be mentioned that the plantlets' root massin all experimental variants has several times exceeded the control.The maximum mass of the corn plantlets treated with cobalt nanoparticles has 2.5 times exceeded the control, in a case with ultrafine humic acids it has been 3 times more and with cuprum nanoparticles 4 times more.The experiment data prove that biologically active nanosubstances promote the plantlets' roots development and this is a necessary condition for their survival at next vegetation stages.
The lab tests have proved that cobalt, cuprum and ultra-fine humic acids nanoparticles have high biological activity.That is why we have chosen for field tests the active agents concentrations having maximum efficiency and these are 0.1 g, 0.5 g and 1.0 g per hectare seeding rate.We have also determined peroxydase, superoxide dismutase (Table 8) and catalase (Table 9) activity during corn vegetation process.Peroxydase activity in corn roots treated with cobalt nanoparticles at maximum concentration has exceeded the control by 29.5%.And superoxide dismutase activity in roots has been 9.9% lower than that of the control.
Humic acids have increased peroxydase activity in corn roots on average by 16%.This ferment activity in leaves has been higher than the control by 2.3 % at concentration of 0.1 g and by 15 % at concentration of 1.0 g.Superoxide dismutase activity has not differed considerably from that of the control.
Cuprum nanoparticles in all doses have also increased peroxydase activity in roots of the experiment plants by up to 26% as compared with the control.At concentrations of 0.1 g and 0.5 gperoxydase activity in leaves has been 0.8% and 8.6% higher than the control correspondingly.Superoxide dismutase activity at concentration of 0.1 g has been 8.2 % higher in roots than those of the control and 9.1% higher in leaves.With the increase of the active agent concentration superoxide dismutase activity has decreased.Catalase activity in corn is within acceptance limits.When using cobalt nanoparticles catalase activity in roots exceeds the control by 2.6 -8.1%.We have noticed this ferment activity true increase in corn leaves only at concentration of 0.1 gby 5.1% as compared with the control.Catalase activity increase shows large amount of metabolic hydrogen peroxide but the experiment plants have not got any considerable activity increase.At maximum concentrations of humic acids and cuprum nanoparticles (1.0 g) catalase activity in corn roots exceeded the control one by 7.5% and 10.7% correspondingly.Catalase activity in the herbage has been lower than the control at cuprum nanoparticles concentration of 1.0 g by 9.1%.
Table 10 shows that corn treated with cuprum nanoparticles have considerable 61 % increase of its amount in seeds and flowers as compared with the control but the leaves practically do not have any changes.Leaves are the place where basic processes connected with energy metabolism take place so cobalt increase has strengthened the biochemical processes.The leaves when the seeds treated with cobalt nanoparticles have got 26 % less ferrum than those of the control.
We have determined the yield (Table 11) and chemical composition of corn seeds ( The use of cuprum nanoparticles at concentration of 0.1 ghascaused 0.43 % fat increase as compared with the control.Cobalt nanoparticles at the same concentration have got 0.63 % fat increase in comparison with the control plants. Total ash characterizes seeds mineral metabolism.Cuprum nanoparticles at concentration of0.1 ghave caused total ash decline in seeds by 0.05% but with the increase of concentration total ash has increased up to the positive difference with the control (+0.29%).Cobalt nanoparticles at concentration of 0.5 ghave caused total ash increase in seeds by 0.2% in comparison with the control.This can prove the more intensive absorption of mineral substances by the plant.And the dry matter of the experiment plants in all variants has been higher than that of the control.
The lab tests with corn seeds and sprouts have proved biological activity of nano materials being studied, i.e. cobaltand cuprum nano particles and ultra fine humicacids.The study of vital, morphological and physiological parameters has shown that the most efficient concentrations are in the range of 0.1-1.0g per hectare seeding rate.
The field tests to study the influence of nanomaterials on corn in vegetation have shown that nanoparticles increase field germination of seeds, growth and development of plants, leaf area and photosynthesis efficiency; strengthen redox enzyme activity and mineral metabolism parameters.Nanoparticles increase the yield of corn seeds and change their chemical composition as for fat, total ash and protein increase.
On the whole, as a result of lab and field tests we can say that cuprum nanoparticles at concentration of 0.1 g per hectare seeding rate are most efficient to increase corn yield and quality.

Table 1 .
The Scheme of Lab and Field Tests

Table 3 .
Morphological Parameters of Corn Seeds when Treated with Biologically Active Nanoparticles

Table 8 .
Peroxydase Activity (inopt.dens.units / g raw material • sec) and Superoxide DismutaseActivity (in act.stand.units / g raw material) in Corn Roots and Leaves Influenced by Different Nanoparticles

Table 9 .
Catalase Activity in Corn Plants and Roots

Table 10 .
Biogenic Metals in CornSeeds treated with cuprum nanoparticles do not have any changes of cobalt.Seeds treated with cuprum nanoparticles have 51 % more ferrum in seeds and flowers than those of the control and it can be connected with biological antagonism of ferrum and cuprum in biochemical processes of plants.The leaves of experiment plants have 18.8 % less ferrum.Corn treatment with cobalt nanoparticles has caused 119 % cuprum increase in leaves in comparison with the control ones.The leaves have also had 200 % more cobalt than the control variant.But flowers and seeds have got 45 % less cobalt.