Project's information

Project's title Research and application of nanotechnology in agriculture
Project’s code VAST.TĐ.NANO.NN/15-18
Research hosting institution Institute of Environmental Technology
Project leader’s name Assoc.Prof. Nguyen Hoai Chau
Project duration 01/03/2015 - 31/12/2019
Project’s budget 56 million VND
Classify Excellent
Goal and objectives of the project

Effectively application of nanotechnology to improve productivity, quality, cost reduction and resistance increase against adverse conditions of weather, soil and epidemics in the production of key agricultural products in Vietnam. Development of research and application of nanotechnology in agriculture through links and cooperation between research institutions in the fields of chemistry, materials, biology, cultivation, animal husbandry and fisheries for agricultural development. Promote the training of scientific staff and publish research results in Vietnam and foreign journals of sciences and technologies on nanotechnology research and application.

Main results

8.1.    Theoretical results:
8.1.1. Set up technological process to produce  nano products for agricultural applications
- Has obtained scientific data related to manufacturing nanomaterials such as the effect of metal nano concentration, stabiliz er concentration, reducing agent ratio, particle size, dispersion ... as the basis to set up technological process of nano preparations used in agriculture. Has obtained scientific data on the effect of nanomaterials on microorganisms such as the ability to inhibit bacteria and fungi that cause diseases in plants and animals, stimulating the plant growth  ...
           - A set of technological processes to produce products used in agriculture has been built. It is noteworthy that the trace element nanoparticles in humic chelate form of foliar fertilizer have been successfully produced to increase the fertilizer efficiency  and copper-silver alloy nanoparticles have also been maked to use for controlling  plant diseases in a more effective way compared to existing plant protection chemicals
8.1.2.Research and application of nanotechnology in crop production
- In maize production: Based on the impact assessment of single metal nanoparticles and its combination were identified combination of nano nutritious microelements used in maize seed treatment (priming) and foliar spraying to stimulate germination and obtain better yield. Maize cultivation protocols have been established to apply nano preparations for seed treatment and foliar fertilizer for the Mekong Delta region and the South Central Coast,  Southeast region. In the Mekong Delta, the protocol allows for a 25% reduction of chemical fertilizers but the yield is equal to or higher than that of the controle. The contents of Cadimi, Lead and Mercury in maize kernels produced by protocol are within the safety threshold when used as human food and animal feed and starch and crude protein content equal to controle. In the South Central Coast and Southeast region, the protocol allows for a 20% reduction in chemical fertilizer compared to the current practice, a slight increase of maize yield and economic efficiency up 17.8-26.5% higher than controls.
Technical protocol of maize cultivation using nano preparations in Mekong Delta has been submitted to Ministry of Agriculture and Rural Development for approval to be the advanced technical protocol at national level  
-In soybeen cultivation: Identified the appropriate formula and dosage of nano preparatiocultivationns for soybean seed treatment and foliar spraying to make soybean grow better, give higher yields. Soybean cultivation protocols using nano preparations in seed treatment and foliar spraying for some northern provinces, Southeast and Mekong Delta region have been established. In some northern provinces, the protocol allows for a 10-30% reduction of macronutrient fertilizer, a 11.4-5.7% yield increase, higher net interest while the quality of soybeen and heavy metal residues in soil are not affected. In the southeast and the Mekong Delta region, the application of protocol help to increase the yield up 14-20% and increase profits. Using nano preparations at the recommended concentration increases the nitrogen fixation capacity of soybean and without affecting the Rhizobium nitrogen fixing bacteria.
-For black pepper, coffee and dragon fruit production :
Determined the appropriate concentrations of Cu, Ag and Zn metal nanoparticles in controlling some important diseases of black pepper, coffee and dragon fruit tree at the nursery stage as well as seedlings.
At the basic construction stage of pepper, the combined-nanoparticles foliar fertilizer of Cu, Zn and B  with dosage of 38.4-43.2 g / ha / year increased the growth parameters while at business peroid,  dosage of 47.0g/ha/ year increased productivity up 12.9-14.2%, increased economic efficiency from 19.8 to 22.9% compared with local farming practices. With coffee at basic construction stage, combined-nanoparticles of Cu, Zn and B  with dosage of 30.7g / ha / year increased growth and dosage of 38.3 g / ha / year suitable for business peroid , increased productivity up 11.1-14.3%, increased economic efficiency from 14.5 to 19.5%. With dragon fruit, the appropriate dosages are 17.8 g / ha / year and  25.0g / ha / year for the basic construction stage and business peroid respectivelly. Dragon fruit productivity and economic efficiency at business peroid  are 12.6-13.6% and 16.8 to 18.2%  higher than the control.
-  Application of nano products in controling important fungal diseases on fruit trees  before and after havesting
The efficacy of nano products in inhibiting some fungi causing rot and droping fruits (C. gloeosporioides and Phytophthora spp) under in vitro, in vivo, nursery and field conditions has been evaluated. The results showed that Cu-Ag alloy nanoparticles had high efficacy equivalent to that of plant protection chemicals, but the dose used was much lower (25 ppm compared with 400 ppm of Carbendazim and Hexaconazole).
A protocol using Cu-Ag alloy nanoparticles (25ppm) in preventing fungi C. gloeosporioides and Phytophthora spp. on oranges before harvest with preventive eficacy of 78.35% and 75.74%, respectively, reduced  friut drop by 76.20% (equivalent to pesticides), increased profits of orange growers to 22 % compared to the control. There is no silver metal residue on orange peel and copper metal residue within the permitted level in fruit and vegetable products.
The protocol using Cu-Ag alloy nanoparticles (50 ppm) was established to control some typical fungi on oranges / mangoes after harvest. The protocol helps to minimize the number of rot fruit, reduce the rate of loss compared to controle, maintain the fruit quality
8.1.3. Research and application of nano-products in pig, cow and shrimp breeding
- Experimental results showed that it’s possible to replace inorganic minerals by nano minerals with appropriate doses in the diet without affecting the their reproductive performance of sows, reducing  incidence of diarrhea of weaned piglets and increase the growth while reducing the amount of feed needed of growing pigs
At the fattening stage, a supplement of a mixture containing nano iron (600 µg), copper (150 µg), cobalt (1.5 µg) and selenium (1.5 µg) per kg of body weight for cows helps better feed intake and increasing body weight gain without changing their blood physiological indicators.
- At the lactation stage, a supplement of nanoparticles Fe (100 - 200 µg), Cu (50 - 100 µg), Co (25 - 50 µg) and Se (10 - 20 µg) per kg of body weight helps improving feed intake, increasing milk production and reducing feed consumption per kg of milk.
- For young calves of 60 - 120 days old, a supplement of Fe (50 - 300 µg), Cu (25-250 µg), Co (12.5-75 µg) and Se (5-30 µg) per kg of body weight helps them grow quicker. This also reduces the number of Salmonella bacteria in their digestive tract and prevent diarrhea.
- Preventing and treating reproductive diseases in cows with 150ppm of nano Ag solution showed a higher efficiency of 2-3 times compared to a treatment with 100-150ml each time.
- Sprays solution for cows’ nail has a preventive effect of 93.33%, which is 13.33% higher than using iodine.
- Nano antibiotics complex Alg-Doxycyclin-TiO2-Ag  & Alg-Doxycyclin-OTC-TiO2-Ag have a good effect against V. cholerae, V. parahaemolyticus and V. alginolyticus
- Shrimps grew better both in their body length and body weight in the experimental ponds using Alg-Doxycyclin-TiO2-Ag compared to the control ones. Their survival rate in experiment was also higher at 80.83% compared to the control group at 70.66%. There were 62 shrimps/kg (experiment) compared to 68 shrimps/kg (control). The productivity reached 4332-5446 kg/pond corresponding to the yield of 17.3 - 21.8 tons/ha.
- No antibiotic residues were recorded in shrimps of experiment groups, while the heavy metal content remained at the allowed threshold. Feed consumption ratio was relatively low at 1.07 for experiment ponds compared to 1.18 for the control ponds.
- First results showed that the profit is doubled in the experimental pond comparing to the control one.
8.1.4. Study on the effect mechanism and biosafety of nano products in the project
    - In germination stage, the positive effects of metal nanoparticles (nFe, nCu, nCo) and metal oxide nanoparticle (nZnO) on germination process of DT26 soybean variety were determined. The analyzed parameters such as germination percentage, germination rate, growth of primary roots and leaves in the soybean seed treated with the above nano particles were higher than those in the control. The enhancement of primary root length has been proved by the increase of root mitotic index in primary root cells.  The most suitable concentration of 4 types of nano particles for treatment of DT26 soybean seed is: nFe 50 mg/L, nCu 50 mg/L, nZnO 50 mg/L and nCo 0.05 mg/L. Among the studied nano particles (NP), nFe exhibited the best effects for germination under normal condition; nZnO showed the best improvement for germination under drought condition. The treatment of soybean seeds with the above NP at tested concentrations does not affect to morphology and development of primary soybean seedlings. At the molecular levels (DNA, gene, protein), the results showed that NP altered the global DNA methylation level of soybean genome, especially at 18 hour- and 48 hour- primary roots  in  germination stage. Moreover, NP treatments lead to the up-regulation of genes involved in ethylene biosynthesis and catabolism of reserves (protein, lipid, carbohydrate), thus increasing germination process of soybean seeds. The above positive effects NP were extended to growth stages (VE, VC, V1, V2, V3 and V4) of soybean seedlings. Especially, seed treatment with NP at suitable concentrations enhanced the drought tolerance at both germination stage and growth stage (V4) of soybean plants. These effects have been proven by a range of adapted properties of morphology (i.e., root length) and physiology (water storage content, drought tolerant index, biomass reduction) of V4 soybean plants under drought condition. Among the studied NP, iron NP showed the best results for DT 26 soybean variety under drought condition. Moreover, molecular analysis of drought samples has showed the higher expression of several drought-responsive genes, which important in the stress signaling pathways, as well as increasing anti-oxidant enzyme in plant cells. Based on all results, we have proposed a hypothesis model of NP acting mechanism in soybean, which can explain to the positive effects of NP in germination, growth and drought tolerance in soybean.
  - Positive effect of nanoscale zerovalent cobalt (NZVC) has been determined (single form, at doses of 0.17 and 16.67 mg/kg of seed) as well as mix of nanoparticle types (0.5 and 2.5 mg/kg of seeds) on the growth indices of cultivars DT26 and DT51. All growth parameters such as plant height, root length, dry weight of stems, leaves and leaf area were higher (4 - 38%) compared with that of the control (no treatment with NZVC). Nanoscale zerovalent cobalt from Vietnam exhibited similar efficiency compared to those from USA. There were no significant differences between doses of 0.17 and 16.67 mg of NZVC/kg of seeds and mix of nanoparticle types. Therefore, we recommended to use NZVC produced by Vietnam at the dose of 0.17 mg/kg of seeds to save and increase economic efficiency. The change of the pigment content in soybean leaves has been determined at different growth stages. The concentration of chlorophyll a and b were tended to increase and reached maximum at fifth (V5) trifoliate stage. This value of the experimental formula (treated with NZVC) was higher than that in the control formula. Photosynthetic parameters expressed by Pn (net photosynthetic rate), Fm (maximal fluorescence), Fo (initial fluorescence), Fv/Fm (efficiency of use light energy absorbed in photosystem II used in photochemical reactions), ETR (photosynthetic electron transport rate), PSII (effective quantum yield of photosystem II) at different stages in soybean leaves showed that the treatment with NZVC helped to improve the efficiency using absorbed light energy by photosystem II, increase the photosynthetic efficiency and maintain photosynthetic machinery’saction when the soybeen growth was decreased. This  contributed to improve productivity (5-7 % compared to the control) as well as grain quality.The analysis of the expression levels of the major genes involved in the structure and functions of photosynthetic apparatus showed that the expression levels of these genes varied according to the growth stages and NZVC concentration. The psaA, Lhca,Cytb6f (belonging to photosystem I) and the psbA (belonging to photosystem II) of the experimental formula tended to increase gradually during the growth stages. The genes coding for photosystem II’s proteins such as psbB, psbC, psbD, psbE and psaB gene of photosystem I tended to increase in expression levels at 20 and 70 days and reduced their expression levels at 40 days. High concentration of NZVC (16.67 mg/kg of seed) have not negatively affected on the photosynthetic function of treated soybean plant. The activities of nitrate and nitrite reductase of soybeans being treated with NZVC were increased continuously over time and reached a higher value than that of the control formula. This demonstrated that the treatment with NZVC improved the nitrogen fixation, conversion and uptake in soybean plants. Treatment soybean seeds with NZVC has caused slight "oxidative stress" in soybean plant DT26 and increased in the activity of antioxidant enzymes such as SOD (superoxide dismutase), CAT (catalase), POX (Peroxidase) and APX (ascorbate peroxidase). This is considered as a protective mechanism of the organisms to reduce these harmful effects caused by oxidative stress. We showed preliminarily proposed a hypothesis model of the mechanism of NZVC impact on the physiological and biochemical processes of soybean and maybe help to explain the mechanism to increase in the productivity of soybean in the above conditions.
- Improve the quality of plantlets as well as other products derived from plant cell tissue culture. Ensuring the stability and efficiency of the culture media with the ability to limit the microbial contamination of additional ingredients into the culture medium. Overcoming a number of abnormal physiological and biochemical phenomena of plants in micropropagation.
- All of single metal nanoparticles such as Ag, Co, Cu, and Fe showed positive effects on bar gene transfer into DT22 soybean cultivar. Among them, Ag, Co, and Cu nanoparticles exhibited increased transgenic efficiency. Especially, Ag nanoparticles showed the best effect in the rate of multi-shoots, regeneration rate after screening by glufosinate, rooting rate, and significantly increased efficiency of gene transfering compared with control condition (without nanoparticles) by 0,011 M concentration. Combinations of two types of metal nanoparticles such as Ag-Co, Ag-Cu, and Cu-Co also showed positive effects on bar gene transfer into DT22 soybean cultivar with different levels depending on each stage of culturing. Particularly, Cu-Co (Cu: 0,01M; Co: 0,01M)  and Ag-Co (Ag: 0,011M; Co: 0,01M) combinations exhibited increase the transgenic efficiency by 0.4% and 0.1%, respectively. The Ag-Co-Cu (Ag: 0,011M; Co: 0,01M; Cu: 0,01M) combination only showed positive effect to the multi-shoot and post-selective regeneration stages. While this combination exhibited a lower transgenic efficiency than control condition (without nanoparticles). The Ag-Co-Cu-Fe combination showed only positive effect to the multi-shoot stage (Ag: 0,05mM, Co: 0,0004mM, Cu: 0,004mM, Fe: 0,5mM) and post-selective regeneration stage (Ag: 0,025mM, Co: 0,0002mM, Cu: 0,002mM, Fe: 0,25mM). But transgenic efficiency did not change when compared with control condition (without nanoparticles).
- Assessed the safety of maize and soybean when using metal nano products for treating seeds before sown; Assessed the safety when using metal nano products as foliar fertilizers for vegetables; Assessed the safety of metal nanoparticles on animal cell lines; Assessed the safety of animal feed supplemented with metal nano products on animals (rabits and pigs).
- Metal nanoparticles and metal oxide nanoparticles (ZnO, Cu, Fe, MnO2, MoO3 and B2O3) at dose of 1µg/g of soil did not reduce the number and types of microorganisms (aerobic bacteria, total aerobic actinomycetes, fungi, phosphate solubilizing microorganism, cellulose decomposing microorganisms and aerobic nitrogen fixing microorganisms); did not reduce the activity of soil enzyme groups (urease, dehydrogenase, acid and alkaline phosphatase and β-D glucosidase) and did not kill Steinernema loci nematodes. Metal nanoparticles and metal oxide nanoparticles (ZnO, Cu, Fe, MnO2, MoO3 and B2O3) applied at dose of greater than 50µg/g of soil caused reduction of the diversity of some microorganism groups (total aerobic bacteria, total aerobic actinomycetes, total fungi, phosphate solubilizing microorganism, cellulose decomposing microorganisms and aerobic nitrogen fixing microorganisms). Applying foliar fertilizer containing metal nanoparticles and nano metal oxides at concentrations of ZnO 350 mg/L; Cu 350 mg/L; Fe 525 mg/L; MnO2 450 mg/L; MoO3 25 mg/L; B2O3 250 mg/L with dose of 0.2 L/ha did not affect the population number and number of species of soil microorganisms (total aerobic bacteria, total aerobic bacteria, total fungi, phosphate solubilizing microorganism, cellulose decomposing microorganisms and aerobic nitrogen fixing microorganisms); reduced soil enzyme activity (urease, dehydrogenase, acid and alkaline phosphatase and β-D glucosidase) and reduced the number of nematodes in the soil. Soil after having used metal nanoparticles and nano metal oxides containing fertilizer at a dose of 0.2 l / ha showed no difference in metal residues, pH, EC, total N, total phosphorus, Ca and Mg when compared to control (fertilizer without nanoparticles).
8.2. Applied results:
          8.2.1. Set up technological process to produce  nano products for agricultural applications
    The produced nano materials that have been tested in experimental pilot agricultural production are highly effective and are now being tested on a large scale by businesses to receive the technology to produce all kinds of products: silver nano skin cream for dairy cows and nano silver solutions for livestock disinfection, foliar fertilizers based on humic nanochelates containing microelements, copper nanoparticles for antifungal plant protection.
    8.2.2.Research and application of nanotechnology in crop production
The protocols using nano products of the project have been established in the cultivation of a number of high economic value crops  in agriculture to increase economic efficiency, reduce costs and reduce environmental pollutions.
- The maize cultivation protocol using nano products for seed treatment and foliar spraying  to help increase economic efficiency and productivity while reduced chemical fertilizers. Protocol applyed for Mekong Delta and South Central Coast regions
- Protocol using nano products (seed treatment and foliar fertilizer) for soybean production in some northern provinces, Southeast and Mekong Delta regions.
        - The Protocols using combined-nanoparticles foliar fertilizer of Cu, Zn and B instead of traditional micronutrients in production of pepper, coffee and fruit dragon in the South
      - The protocols  using Cu-Ag nano alloy products to prevent diseases (fruit rot and drop )  on orange caused by fungi C. gloeosporioides and Phytophthora spp.
       - The protocol  using Cu-Ag nano alloy products  in the control of some typical fungi on oranges and mangoes post-harvest
8.2.3. Research and application of nano-products in pig, cow and shrimp breeding
The  research results showed the effectiveness of the nano complex in pigs, cows and shrimp breeding. Therefore, nano-products are considered a promising material for use in dairy and beef cattle in particular and for ruminants in general as well as in aquaculture. Nanoparticles applied in this work have reduced the impact on the environment, and also brought health and high efficiency.
    8.2.4. Study on the effect mechanism and biosafety of nano products in the project    
- The obtained results are provided the scientific basic for employing studied NP in agriculture. The treatment of  studied nano particles, especially, iron NP with concentration at 50 mg/L (equivalent to dose 17.5 mg/kg seeds) and cobalt NP with  concentration at 0.05 mg/L (equivalent to dose 0.0175 mg/kg seeds) can be used as a very effective approach for enhancement of germination rate, growth and drought tolerance of soybean and other crops. The used concentrations were very low, economical but highly effective.
- The suitable dose of nanoscale zerovalent cobalt for the treatment of soybean seeds was 0.17 mg/ kg of seeds. This concentration was low, economical but having high effective. The obtained results of the second branch project provided the scientific basis for implementing studies using nanoscale zerovalent cobalt in a larger scale in practical production.
- Silver nanoparticle can be applied in transgenic procedure on DT22 soybean cultivar through Agrobacterium tumefaciens.
- Research is novelty, high scientific content as well as high applicability, and can contribute to the improvement of productivity and quality of micropropagation products in production practices.

Novelty and actuality and scientific meaningfulness of the results
  • Set up technological process to produce  nano products for agricultural applications

Engineered nanomaterials  have been produceded and successfully applied in cultivation and animal husbandry, typically:
- Nano products for seed treatment of maize and soybeen before sowing
- Trace elements - containing foliar nanofertilizers for maize, soybean, coffee, pepper..
           - Copper-silver alloy nanoparticles to prevent fungal diseases  caused fruit rot and drop on oranges before and after harvesting;
- Nanosilver cream used in dairy farming
- Nanosilver solution used for disinfection in animal husbandry
- Multi-functional nano preparations carrying antibiotics to prevent acute hepatopancreatic necrosis disease in White Leg shrimp (Litopenaeus vannamei)

  • Research and application of nanotechnology in crop production
  • Research and application of nano-products in pig, cow and shrimp breeding
  • Study on the effect mechanism and biosafety of nano products in the project    

- The obtained results  are provided the scientific basic for employing studied NP in agriculture. The treatment of  studied nano particles, especially, iron NP with concentration at 50 mg/L (equivalent to dose 17.5 mg/kg seeds) and cobalt NP with  concentration at 0.05 mg/L (equivalent to dose 0.0175 mg/kg seeds) can be used as a very effective approach for enhancement of germination rate, growth and drought tolerance of soybean and other crops. The used concentrations were very low, economical but highly effective.
- The suitable dose of nanoscale zerovalent cobalt for the treatment of soybean seeds was 0.17 mg/ kg of seeds. This concentration was low, economical but having high effective. The obtained results of the second branch project provided the scientific basis for implementing studies using nanoscale zerovalent cobalt in a larger scale in practical production.
- Silver nanoparticle can be applied in transgenic procedure on DT22 soybean cultivar through Agrobacterium tumefaciens.
- Research is novelty, high scientific content as well as high applicability, and can contribute to the improvement of productivity and quality of micropropagation products in production practices.

Products of the project

- International papres (10)
- National papers (59)
- Patents (06):
       1- Utility solution. Cream incorporated with silver nanoparticles  in prevention and treatment of mastitis in dairy cows. Utility solution patent No 2002 granted by decision No 18291/QĐ-SHTT dated March 12th, 2019. Institute of Environmental Technology  
      2-  Invention. Preparations for processing seeds before sowing. Application No 1-2016-04592 according to the Decision accepting the application No 84595/QĐ-SHTT dated  December 16th ,2016. Institute of Environmental Technology
    3- Invention.  Method of producing Ag-Cu / Silica nanocomposite materials to treat hoof inflammatory diseas in dairy cows  and materials obtained from this method.
Application No. 1-2018-02629 according to the Decision accepting the application No. 50747 / QD-SHTT dated July 18th, 2018. Institute of Environmental Technology
4- Invention.  Process of production of  nanochelat foliar fertilizer of trace elements using electrochemical activationed catolite as a reaction medium and the preparations obtained from this process.  Application No 1-2018-00571 according to the Decision accepting the application No 8932/QĐ-SHTT dated Fabruary 12th ,2018. Institute of Environmental Technology
5-Invention. The process of preparing antibacterial preparations containing nano silver, nano chitosan and hydrogen peroxide used in veterinary and antibacterial preparations obtained from this process. Application No 1-2018-03786 according to the Decision accepting the application No  68172/QĐ-SHTT dated September 28th , 2018. Institute of Environmental Technology
6- Invention. Method of making Doxycyclin nano antibiotic materials. Application No 1-2019-01324 according to the Decision accepting the application No  34903/QĐ-SHTT, datef May 7th ,2019. Institue of Material Sciences
    - Other products (if applicable):
Project has trained 14 master students, 04 Ph.Dcandidates and many
undergraduate students

Research area

The project's products have been tested and applied in practice in many  provinces such as Hanoi, Thai Nguyen, Thanh Hoa, Thua Thien-Hue, Binh Dinh, Quang Ngai, Ninh Thuan, Dong Nai, Dak. Lak, Gia-Lai, Binh Phuoc, Hau Giang, Dong Thap, Vinh Long ...
       - To apply the protocols using nano products in maize production created by project in the Mekong Delta and South Central Coast region, for soybean production in some northern provinces, Southeast and Mekong Delta region, for cultivation of black pepper, coffee and Dragon fruit in the South and for orange cultivation in the northern provinces of VN
        - Enterprises and laboratories can change their approach to new methods of using nanomaterials in research and production as well as through their ability to overcome some abnormal phenomena encountered by in-vitro plants, can help improve seedling quality and labor productivity (reduce wastage) ...
       - To Suggest the application of nano Ag in gene transformation experimence into DT22 soybeens through Agrobacterium tumefaciens bacteria.

 

Recommendations

      - To continue researching to perfect, improve and expand the scale of manufacturing nanomaterials; continue to research and build a manufacturing process of nano products in an organic and environmentally friendly manner.
    - To Continue  research and application of nano products for Animal husbandry àn cultivation  according to intensive manner  such as using preparetions corresponding to each stage of growth and development.
     -To perform further research of interaction and accumulation of NP within plant cells. The positive effects of studied nanoparticles on growth, development and drought tolerance of plants are depended on types of NP and doses of treatment, Therefore, it is important to carry out the evaluation of type NP and dose treatment which suitable to each crop in field trials.
       -To continue research on: Research on molecular mechanism of cobalt nanoparticles impacting on photosynthetic activity in soybean plants by assessing the level of protein expression - products of genes related to the structure of the photosynthetic apparatus such as subunits of CP43, CP47, cytochrome electronic transport complex, light harvesting complex. Research on the interaction between cobalt and ion transport channels in cell membranes and the balance of essential nutritional factors inside plants