Source: CONNECTICUT AGRICULTURAL EXPERIMENT STATION submitted to
NANOSCALE ELEMENTS SUPPRESS PLANT DISEASE, ENHANCE MACRONUTRIENT USE EFFICIENCY, AND INCREASE CROP YIELD
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1009029
Grant No.
2016-67021-24985
Project No.
CONH00146
Proposal No.
2015-07766
Multistate No.
(N/A)
Program Code
A1511
Project Start Date
Mar 1, 2016
Project End Date
Dec 26, 2019
Grant Year
2016
Project Director
White, J. C.
Recipient Organization
CONNECTICUT AGRICULTURAL EXPERIMENT STATION
PO BOX 1106
NEW HAVEN,CT 06504
Performing Department
Analytical Chemistry
Non Technical Summary
Theprimary goal is to use nanoscale elements as a comprehensive agricultural amendment strategy to suppress crop disease, improve nutritional content, and enhance overall yield. Soil pathogens significantly limit agricultural production, reducing crop yield by 10-20% and resulting in billions of dollars in annual losses. Separately, pathogen control efforts by fungicides exceed $600 million per year in the US. At the same time, macronutrient utilization remains low due to environmental losses and low availability. This shortfall in food production will worsen with a changing climate and an increasing population. Nanotechnology can play a critical role in maximizing global food production and achieving food security. Current approaches have focused on nano-enabled conventional agrichemicals, nanosensors, and waste treatment strategies. However, little is known about the effects of nanoparticle (NP) elements on disease suppression, macronutrient uptake, and crop growth. For example, plant micronutrients are pivotal in disease resistance through activation of defense barrier production and by affecting the systemic acquired resistance pathway. Unfortunately, element availability in soil is limited and foliarly applied micronutrients are not significantly translocated to roots. Although NP intra plant translocation has been reported, there is no information on whether enhanced translocation of NP elements can deter root or shoot pathogens. Based on preliminary data and a recent review of the literature, our central hypothesis is that NP elements can be used to strategically suppress disease, improve nutritional status and enhance crop growth and yield. Our four objectives are:Obj.1- Demonstrate NP efficacy upon foliar or root application at suppressing fungal pathogen infection in model vegetable and grain species.Obj.2- Determine the role of soil type and NP source in disease suppression and crop yield.Obj.3- Determine the impact of NP element treatment on macronutrient (NPK) utilization.Obj.4- Characterize NP absorption and translocation mechanisms in plant tissues by scanning/ transmission electron microscopy (S/TEM-EDS).Using greenhouse and field trials, we will grow tomato, eggplant, corn, and squash in pathogen (Fusarium, Verticillium, Sphaerotheca fuliginea) infested areas. Foliar or root applications of NP metal oxides (CuO, MnO, ZnO, CeO2) will be made at the transplant stage. Bulk and ion controls will be included. A broad range of NP application rates will be used to quantify the dose-response nature of disease suppression or of macronutrient uptake efficiency. Measured parameters include biomass/yield and disease severity; plant tissues will be analyzed for element/nutritional content using ICP-MS and S/TEM-EDS, which also enables an assessment of risk from any residual NPs.Although NP use in agriculture is increasing, the stresses from a changing climate and increasing population require novel strategies for enhancing food production. The use of NP metal oxides to suppress disease and promote yield has great potential to maximize agricultural output. The multidisciplinary team of investigators will establish a Center of Excellence that deploys a unique combination of educational and dissemination strategies to engage a range of stakeholders.
Animal Health Component
0%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
20501101150100%
Knowledge Area
205 - Plant Management Systems;

Subject Of Investigation
0110 - Soil;

Field Of Science
1150 - Toxicology;
Goals / Objectives
Theprimary goal of this project is to use nanoscale elements as a comprehensive agricultural amendment strategy to suppress crop disease, improve nutritional content, and enhance overall yield. Soil pathogens significantly limit agricultural production, reducing crop yield by 10-20% and resulting in billions of dollars in annual losses. Separately, pathogen control efforts by fungicides exceed $600 million per year in the US. At the same time, macronutrient utilization remains low due to environmental losses and low availability. This shortfall in food production will worsen with a changing climate and an increasing population. Nanotechnology can play a critical role in maximizing global food production and achieving food security. Current approaches have focused on nano-enabled conventional agrichemicals, nanosensors, and waste treatment strategies. However, little is known about the effects of nanoparticle (NP) elements on disease suppression, macronutrient uptake, and crop growth. For example, plant micronutrients are pivotal in disease resistance through activation of defense barrier production and by affecting the systemic acquired resistance pathway. Unfortunately, element availability in soil is limited and foliarly applied micronutrients are not significantly translocated to roots. Although NP intra plant translocation has been reported, there is no information on whether enhanced translocation of NP elements can deter root or shoot pathogens. Based on preliminary data and a recent review of the literature, our central hypothesis is that NP elements can be used to strategically suppress disease, improve nutritional status and enhance crop growth and yield. Our four objectives are:Obj.1- Demonstrate NP efficacy upon foliar or root application at suppressing fungal pathogen infection in model vegetable and grain species.Obj.2- Determine the role of soil type and NP source in disease suppression and crop yield.Obj.3- Determine the impact of NP element treatment on macronutrient (NPK) utilization.Obj.4- Characterize NP absorption and translocation mechanisms in plant tissues by scanning/ transmission electron microscopy (S/TEM-EDS).
Project Methods
Obj. 1: Greenhouse and field trials will be conducted to assess NP element efficacy for suppressing disease for 4 crops. A determination of particle toxicity to the plants will also be made. Healthy transplants of eggplant, tomato, squash and corn will be produced from seed in a greenhouse. These crops are known to be susceptible to fungal pathogens. When seedlings are at the 2 or 4 leaf stage, plants will be removed from the potting mix and the roots will be washed. For the foliar treatments, the shoots tissue will be immersed in each treatment at rates of 0-500 mg/L of the elements CuO, MnO, ZnO, or CeO2. The element solutions will be prepared separately as NP, bulk or chelated materials. Plants will be placed on paper towels until leaves are dry and then transplanted into 10-cm pots filled with silica sand (1 plant/pot). For the root amendments, seedlings will be transplanted into 10 cm pots filled with silica sand and then exposed to 50 ml of 0, 10, 100, or 500mg/L of the element solutions described above for foliar application. Plants will be watered as needed with normal fertilizer regiments of 20-20-20 NPK. Growth conditions approximating 25oC with a 14 h photoperiod of 200 µmol m-2 s-1 PAR will be used for the full period of the experiment. There will be 12 replicates for both foliar and root applications, but one week after element metal oxide treatment, half of the plants will be inoculated with disease by drenching the sand with a spore suspension (106 spores/ml dH20) of host specific strains of Fusarium oxysporum for tomato and corn and Verticillium dahliae for eggplant. The uninoculated plants will receive dH20 as control. Since powdery mildew (Sphaerotheca fuliginea) of squash is a foliar obligate pathogen and cannot be cultured, it will be inoculated via naturally infested tissue saved from previous field trials. The fungicide chlorothalonil will be used to provide a healthy (disease free) control plants for this treatment. Disease severity will be monitored weekly after symptom appearance. Fresh and dry weights will be recorded after 6 weeks. Subsamples of foliar tissues and roots will be freeze-dried, ground and analyzed for chlorophyll and total nonstructural carbohydrates (TNC). Field plots will be established at the Lockwood farm in Hamden CT. Plots measuring 3.6 m long and that are 1.2 m apart will be planted with tomato, eggplant, squash or corn based on results above. Seedlings will be treated (roots or shoots) with NP or other solutions as described above in the greenhouse prior to transplanting into uncontaminated potting mix. After an additional 7 days, healthy plants will be transferred to field plots. Soil in these plots will either be infested with the root/shoot pathogens, or transplants will be inoculated at field planting by dipping transplant roots into a spore suspension of each pathogen. Plants will receive standard fertilization regiments and incidental foliar pests will be controlled as needed. Pathogen growth and crop yield will be monitored weekly by visually inspection and disease progression estimation. During growth and at harvest of both greenhouse and field-grown plants, predetermined subsamples of foliar/root tissue of each plant under each treatment (particle type, concentration, disease exposure) will be taken and delivered to either the CAES PD or UTEP CoPD. The plant tissues will be oven dried and digested on a digital programmable hot block in concentrated acid.. After digestion, the total element content of tissues will be determined by inductively coupled plasma optical emission spectroscopy (ICP-OES) or ICP with mass spectrometry (ICP-MS).Obj. 2: Greenhouse trials as described for objective 1 will be established but instead of sand, soils from 4 locations will be evaluated. All 4 soils differ significantly in key agronomic parameters. Three of the soils will be from the three CAES research farms in Hamden, Windsor and Griswold CT. A fourth soil will be used from a research farm in Fabens TX. The experimental approach will be identical to that described in the greenhouse sand-based studies under objective 1. Analysis of disease severity, biomass/yield, chlorophyll and nutritional/element content will occur as described under objective 1 and will be correlated with the key soil properties mentioned above. For select soil-plant/pathogen systems, multiple sources of two separate NPs will be evaluated. Specifically, three separate vendors will be used to acquire NP CuO and MnO. All materials will be thoroughly characterized at CAES and UTEP for particle size and zeta potential Based on data from objective 1, soil-plant/pathogen systems where nanoscale effects were particularly robust will be selected and amendment experiments will be conducted as above with the CuO and MnO NPs acquired from the different sources. Observed differences in disease suppression, biomass/yield, physiological parameters and element content will be correlated with particle source and properties.Obj. 3: Greenhouse studies will evaluate the influence of NP amendment on the use efficiency of NPK fertilizers by tomato and corn (in the absence of pathogens). This will involve greenhouse trials at the VFRC facility using a locally acquired agricultural soil frequently used in other work. Two macronutrient (NPK) fertilizer rates will be tested with the NPs showing the most promise to enhance crop yield. Based on preliminary data, NP ZnO and MnO will be used. This will be compared with conventional elements of each. Plants will be grown in 8-liter pots, with the NPK fertilizers applied at sowing by soil amendment with scaled amounts per pot. Four weeks after germination, the plants will be divided into two groups, and one sub-group will be soil-treated while the other is foliar-treated with the conventional or NP elements at 100 mg/kg. The study will involve 8 replicates per treatment. Midway through the plant growth, four plants of each treatment will be destructively sampled for biomass and shoot element/nutrient content; the remaining plants will be sampled at the end of the study for biomass, yield and nutrient content in grains/fruit. For the foliar treatment, shoot will be washed thoroughly to eliminate surface adsorbed NPs prior to preparing samples for ICP-MS. The use of high and low NPK rates will provide information on whether use of NP elements can lead to the reduction in the amount of macronutrient input required, and if so, whether there are differences between NPs and salt forms of the element.Obj. 4: During growth and at harvest, subsamples of foliar and root tissue will be taken and delivered to the CAES PD or to the UTEP CoPD. Thin tissue sections of all appropriate treatments will be prepared using an ultramictrotome and analyzed by scanning/transmission electron microscopy (S/TEM) to identify particle distribution and size within shoot/root samples. Particle elemental composition is then determined with energy dispersive X-ray spectroscopy (EDS). The particle elemental content of sampled tissues will then be correlated with metal oxide treatment, disease progression and biomass/yield data.

Progress 03/01/16 to 02/28/20

Outputs
Target Audience:The primary target audience for this project includes plant pathologists and growers interested in novel methods of suppressing root crop disease and enhancing overall agricultural productivity. State and federal regulators interested in nanotechnology are also a target audience for this project. Last, those concerned with the sustainable use of nanotechnology in agriculture are also a target audience for this project. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Five UTEP graduate students participated in this grant over the course of the project, as well as one post-doctoral researcher at CAES. A technician was supported at IFDC. Graduate students or post-docs gave approximately 20% of the 58 presentations that occurred over the course of the project and were obviously involved in all of 35 publications. How have the results been disseminated to communities of interest?A total of 19 publications were published during the last year, bringing the total to 35 for the whole project. We currently have approximately 8 papers in various stages of writing and/or publication. A total of 58 presentations were given during the project period at a range of domestic (37) and international venues (21).Last, we created a website (http://www.ct.gov/caes/cwp/view.asp?a=4898&q=585400 ) for our USDA NIFA AFRI Center of Excellence entitled The Center for Nanotechnology and Agricultural Pathogens Suppression (CeNAPS). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? A number of studies were conducted and published this final year of the project. Significant findings include the following. In two studies focused on the tuning of particle properties, we showed that copper phosphate nanosheets could effectively suppress root disease in both watermelon and tomato at a dose that was 10 times less than amorphous copper oxide nanoparticles. In the second study, time-dependent transcriptomic analysis demonstrated that this enhanced efficacy was a function or more rapid activation of defense-related genes in the roots as a function of particle characteristics. Notably, non-nanoscale forms of copper had no effect on disease. A third paper on copper oxide nanoparticles should similar beneficial effects in soybean that was suffering from fungal infection (sudden soybean death). In an important study with zinc nanoparticles, we showed that many of the effects experienced in bean, both positive and negative as a function of dose, were not observed in the next generation of plants (seeds and seedlings). These findings are important to demonstrating the safety and sustainability of these types of nano-enabled platforms. Three other studies completed this last year also focused on zinc oxide nanoparticles. Studies with zinc are particularly important given the deficiency of this nutrient in soils globally, which then results in subsequent nutritional deficiencies in humans. In one study, we showed that both fresh and aged nanoscale zinc oxide in soil can modulate the uptake of other macro- and micronutrients by wheat. In the second study, an addition-omission strategy demonstrated that in a mixture of zinc, copper and boron nanoscale micronutrients, zinc was the most important element. Last, exposure of drought-stressed sorghum to nanoparticle zinc oxide in the soil significantly increased the drought tolerance of this plant. Given the likely need to grow crops on more marginal soils in the future, these findings are particularly significant. Last, we published two important critical reviews of this field this year; the first was in Environmental Science: Nano and the second was published in Nature Nanotechnology and was featured on the cover. Across the entire 4 years of the project, all objectives of the project were met and significant knowledge was gained on the use of nanoscale elements to suppress disease, enhance macronutrient use efficiency, and increase crop yield. This has included work done at both the greenhouse and field scale. Much of the work was mechanistic in nature and will provide significant knowledge to others working in this field, guiding future basic research as well as application oriented studies. As with any successful research grant, many questions were answered and many new questions have arisen. We are currently seeking both AFRI and Hatch funds to bring this research to the next level of understanding and technology development. ?

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adisa, I., V.L.R. Pullagurala, S. Rawat, J.A. Hernandez-Viezcas, C.O. Dimkpa, W.H. Elmer, J.C. White, J.R. Peralta-Videa, J.L. Gardea-Torresdey. 2018. Role of cerium compounds in Fusarium wilt suppression and growth enhancement in tomato (Solanum lycopersicum). J. Agric. Food Chem. 66:5959-5970.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Borgatta, J., C. Ma, N. Hudson-Smith, W.H. Elmer, C.D. Plaza P�re, R. De La Torre-Roche, N. Zuverza-Mena, C.L. Haynes, J.C. White, R.J. Hamers. 2018. Copper nanomaterials suppress root fungal disease in watermelon (Citrullus lanatus): Role of particle morphology, composition and dissolution behavior. ACS Sustain. Chem. Eng. 6(11):1484714856.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dimkpa, C.O., I.O. Adisa, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2018. Effect of manganese nanoparticle exposure on nutrient acquisition in wheat. Agronomy 8(9):158, doi.org/10.3390/agronomy8090158.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dimkpa, C.O., U. Singh, P.S. Bindraban, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2018. Exposure to weathered and fresh nanoparticle and ionic Zn in soil promotes grain yield and modulates nutrient acquisition in winter wheat (Triticum aestivum). J. Agric. Food Chem. 66 (37): 9645-9656.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dimkpa, C.O., U. Singh, P.S. Bindraban, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2019. Zinc oxide nanoparticles alleviate drought-induced alterations in sorghum performance, nutrient acquisition, and grain fortification. Sci. Tot. Environ. 688:926-934.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dimkpa, C.O., U. Singh, P.S. Bindraban, I.O. Adisa, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2019. Addition-omission of zinc, copper, and boron nano and bulk oxide particles demonstrate element and size -specific response of soybean to micronutrients exposure. Sci. Tot. Environ. 665:606-616.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ma, C., J. Borgatta, R. De La Torre Roche, N. Zuverza-Mena, J.C. White, R.J. Hamers, W.H. Elmer. 2019. Time-dependent transcriptional response of tomato (Solanum lycopersicum L.) to Cu nanoparticle exposure upon infection with Fusarium oxysporum f. sp. Lycopersici. ACS Sust. Chem. Eng. 7(11):10064-10074.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Medina-Velo, I.A., N. Zuverza-Mena, C. Tamez; Y. Ye, J.A. Hernandez-Viezcas, J.C. White, J.R. Peralta-Videa, J.L. Gardea-Torresdey. 2018. Minimal transgenerational effect of ZnO nanomaterials on the physiology and nutrient profile of Phaseolus vulagaris. ACS Sustain. Chem. Eng. 6:7924-7930.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adisa, I., V.L.R. Pullagurala, J.R. Peralta-Videa, C.O. Dimkpa, C. Ma, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2019. Recent advances in nano-enabled fertilizers and pesticides: A critical review of mechanisms of action. Environ. Sci.: Nano. DOI: 10.1039/C9EN00265K.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Elmer, W.H., C. Ma, J.C. White. 2018. Nanoparticles for plant disease management. Curr. Op. Environ. Sci. Health 6:66-70.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Guo, H., J.C. White, Z. Wang, B. Xing. 2018. Nano-enabled fertilizers to control the release and use efficiency of nutrients. Curr. Op. Environ. Sci. Health 6:77-83.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kah, M., N. Tufenkji, J.C. White. 2019. Nano-enabled strategies to enhance crop nutrition and protection. Nature Nano. 14:532-540.
  • Type: Book Chapters Status: Published Year Published: 2019 Citation: Tan W., Deng, C., Peralta-Videa, J., Gardea-Torresdey, J. L., 2019. Interaction of nanomaterials in secondary metabolites accumulation, photosynthesis, and nitrogen fixation in plant systems. In: Verma, S. K., Das, A. K. (Eds) Analysis, fate, and toxicity of engineered nanomaterials in plants. Comprehensive Analytical Chemistry. Elsevier, chapter 3, pp. 55-74.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Rawat, S., Adisa I. O., Wang, Y., Sun, Y., Fadil, A. S., Niu, G., Sharma, N., Hernandez-Viezcas, J. A., Peralta-Videa, J. R., Gardea-Torresdey, J. L. 2019. Differential physiological and biochemical impacts of nano vs micron Cu at two phenological growth stages in bell pepper (Capsicum annuum) plant. NanoImpact 14, 100161.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Tamez, C., Hernandez, M., Hern�ndez-Viezcas, J. A. Gardea-Torresdey, J. L. 2019. Uptake, Transport, and Effects of Nano-Copper Exposure in Zucchini (Cucurbita pepo). Science of the Total Environment 665,100-106.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Tamez, C., Morelius, E., Hernandez-Viezcas, J., Peralta-Videa, J.R., Gardea-Torresdey, J. 2019. Biochemical and physiological effects of copper compounds/nanoparticles on sugarcane (Saccharum officinarum). Science of the Total Environment 649, 554-562.
  • Type: Book Chapters Status: Published Year Published: 2019 Citation: Pullagurala, V. L. R., Adisa, I.O., Rawat, S., White, J.C., Zuverza-Mena, N., Hernandez-Viezcas, J.A., Peralta-Videa, J.R., Gardea-Torresdey, J.L. (2019). Fate of engineered nanomaterials in agroenvironments and impacts on agroecosystems. In: Marmiroli, N., White, J.C., Song, J. (Eds.) Exposure to Engineered Nanomaterials in the Environment. Elsevier, chapter 4, pp. 105-142 .
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Per�z, C.D.P.; De La Torre Roche, R.; Zuverza-Mena, N.; Ma, C.; Shen, Y.; White, J.C.; Amp�lio Pozza, E.; Pozza, A.A.A.; Elmer, W.H. 2019. Metalloid and metal oxide nanoparticles suppress Sudden Death Syndrome of soybean. J. Ag. Food Chem. doi.org/10.1021/acs.jafc.9b06082.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pagano, L., E. Maestri, J.C. White, N. Marmiroli, M. Marmiroli, M. 2018. Quantum dots exposure in plants: Minimizing the adverse response. Curr. Op. Environ. Sci. Health 6:71-76.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pagano, L., E. Maestri, M. Caldara, J.C. White, N. Marmiroli, M. Marmiroli, M. 2018. Engineered nanomaterial activity at the organelle level: Impacts on the chloroplast and mitochondria. ACS Sustain. Chem. Eng. 6(10):12562-12579.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ye, Y., Medina-Velo, I. A., Cota-Ruiz K., Moreno-Olivas, F., Gardea-Torresdey, J. L. 2019. Can Abiotic Stresses in Plants be Alleviated by Manganese Nanoparticles or Compounds? Ecotoxicology and Environmental Safety 184, 109671.


Progress 03/01/16 to 12/26/19

Outputs
Target Audience:The primary target audience for this project includes plant pathologists and growers interested in novel methods of suppressing root crop disease and enhancing overall agricultural productivity. State and federal regulators interested in nanotechnology are also a target audience for this project. Last, those concerned with the sustainable use of nanotechnology in agriculture are also a target audience for this project. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Five UTEP graduate students participated in this grant over the course of the project, as well as one post-doctoral researcher at CAES. A technician was supported at IFDC. Graduate students or post-docs gave approximately 20% of the 58 presentations that occurred over the course of the project and were obviously involved in all of 35 publications. How have the results been disseminated to communities of interest?A total of 19 publications were published during the last year, bringing the total to 35 for the whole project. We currently have approximately 8 papers in various stages of writing and/or publication. A total of 58 presentations were given during the project period at a range of domestic (37) and international venues (21).Last, we created a website (http://www.ct.gov/caes/cwp/view.asp?a=4898&q=585400 ) for our USDA NIFA AFRI Center of Excellence entitled The Center for Nanotechnology and Agricultural Pathogens Suppression (CeNAPS). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? A number of studies were conducted and published this final year of the project. Significant findings include the following. In two studies focused on the tuning of particle properties, we showed that copper phosphate nanosheets could effectively suppress root disease in both watermelon and tomato at a dose that was 10 times less than amorphous copper oxide nanoparticles. In the second study, time-dependent transcriptomic analysis demonstrated that this enhanced efficacy was a function or more rapid activation of defense-related genes in the roots as a function of particle characteristics. Notably, non-nanoscale forms of copper had no effect on disease. A third paper on copper oxide nanoparticles should similar beneficial effects in soybean that was suffering from fungal infection (sudden soybean death). In an important study with zinc nanoparticles, we showed that many of the effects experienced in bean, both positive and negative as a function of dose, were not observed in the next generation of plants (seeds and seedlings). These findings are important to demonstrating the safety and sustainability of these types of nano-enabled platforms. Three other studies completed this last year also focused on zinc oxide nanoparticles. Studies with zinc are particularly important given the deficiency of this nutrient in soils globally, which then results in subsequent nutritional deficiencies in humans. In one study, we showed that both fresh and aged nanoscale zinc oxide in soil can modulate the uptake of other macro- and micronutrients by wheat. In the second study, an addition-omission strategy demonstrated that in a mixture of zinc, copper and boron nanoscale micronutrients, zinc was the most important element. Last, exposure of drought-stressed sorghum to nanoparticle zinc oxide in the soil significantly increased the drought tolerance of this plant. Given the likely need to grow crops on more marginal soils in the future, these findings are particularly significant. Last, we published two important critical reviews of this field this year; the first was in Environmental Science: Nano and the second was published in Nature Nanotechnology and was featured on the cover. Across the entire 4 years of the project, all objectives of the project were met and significant knowledge was gained on the use of nanoscale elements to suppress disease, enhance macronutrient use efficiency, and increase crop yield. This has included work done at both the greenhouse and field scale. Much of the work was mechanistic in nature and will provide significant knowledge to others working in this field, guiding future basic research as well as application oriented studies. As with any successful research grant, many questions were answered and many new questions have arisen. We are currently seeking both AFRI and Hatch funds to bring this research to the next level of understanding and technology development. ?

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dimkpa, C.O., U. Singh, P.S. Bindraban, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2019. Zinc oxide nanoparticles alleviate drought-induced alterations in sorghum performance, nutrient acquisition, and grain fortification. Sci. Tot. Environ. 688:926-934.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Dimkpa, C.O., U. Singh, P.S. Bindraban, I.O. Adisa, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2019. Addition-omission of zinc, copper, and boron nano and bulk oxide particles demonstrate element and size -specific response of soybean to micronutrients exposure. Sci. Tot. Environ. 665:606-616.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ma, C., J. Borgatta, R. De La Torre Roche, N. Zuverza-Mena, J.C. White, R.J. Hamers, W.H. Elmer. 2019. Time-dependent transcriptional response of tomato (Solanum lycopersicum L.) to Cu nanoparticle exposure upon infection with Fusarium oxysporum f. sp. Lycopersici. ACS Sust. Chem. Eng. 7(11):10064-10074.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Medina-Velo, I.A., N. Zuverza-Mena, C. Tamez; Y. Ye, J.A. Hernandez-Viezcas, J.C. White, J.R. Peralta-Videa, J.L. Gardea-Torresdey. 2018. Minimal transgenerational effect of ZnO nanomaterials on the physiology and nutrient profile of Phaseolus vulagaris. ACS Sustain. Chem. Eng. 6:7924-7930.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adisa, I., V.L.R. Pullagurala, S. Rawat, J.A. Hernandez-Viezcas, C.O. Dimkpa, W.H. Elmer, J.C. White, J.R. Peralta-Videa, J.L. Gardea-Torresdey. 2018. Role of cerium compounds in Fusarium wilt suppression and growth enhancement in tomato (Solanum lycopersicum). J. Agric. Food Chem. 66:5959-5970.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Borgatta, J., C. Ma, N. Hudson-Smith, W.H. Elmer, C.D. Plaza P�re, R. De La Torre-Roche, N. Zuverza-Mena, C.L. Haynes, J.C. White, R.J. Hamers. 2018. Copper nanomaterials suppress root fungal disease in watermelon (Citrullus lanatus): Role of particle morphology, composition and dissolution behavior. ACS Sustain. Chem. Eng. 6(11):1484714856.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dimkpa, C.O., I.O. Adisa, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2018. Effect of manganese nanoparticle exposure on nutrient acquisition in wheat. Agronomy 8(9):158, doi.org/10.3390/agronomy8090158.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dimkpa, C.O., U. Singh, P.S. Bindraban, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2018. Exposure to weathered and fresh nanoparticle and ionic Zn in soil promotes grain yield and modulates nutrient acquisition in winter wheat (Triticum aestivum). J. Agric. Food Chem. 66 (37): 9645-9656.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adisa, I., V.L.R. Pullagurala, J.R. Peralta-Videa, C.O. Dimkpa, C. Ma, W.H. Elmer, J.L. Gardea-Torresdey, J.C. White. 2019. Recent advances in nano-enabled fertilizers and pesticides: A critical review of mechanisms of action. Environ. Sci.: Nano. DOI: 10.1039/C9EN00265K.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Elmer, W.H., C. Ma, J.C. White. 2018. Nanoparticles for plant disease management. Curr. Op. Environ. Sci. Health 6:66-70.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Guo, H., J.C. White, Z. Wang, B. Xing. 2018. Nano-enabled fertilizers to control the release and use efficiency of nutrients. Curr. Op. Environ. Sci. Health 6:77-83.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kah, M., N. Tufenkji, J.C. White. 2019. Nano-enabled strategies to enhance crop nutrition and protection. Nature Nano. 14:532-540.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pagano, L., E. Maestri, J.C. White, N. Marmiroli, M. Marmiroli, M. 2018. Quantum dots exposure in plants: Minimizing the adverse response. Curr. Op. Environ. Sci. Health 6:71-76.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pagano, L., E. Maestri, M. Caldara, J.C. White, N. Marmiroli, M. Marmiroli, M. 2018. Engineered nanomaterial activity at the organelle level: Impacts on the chloroplast and mitochondria. ACS Sustain. Chem. Eng. 6(10):12562-12579.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ye, Y., Medina-Velo, I. A., Cota-Ruiz K., Moreno-Olivas, F., Gardea-Torresdey, J. L. 2019. Can Abiotic Stresses in Plants be Alleviated by Manganese Nanoparticles or Compounds? Ecotoxicology and Environmental Safety 184, 109671.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Tamez, C., Hernandez, M., Hern�ndez-Viezcas, J. A. Gardea-Torresdey, J. L. 2019. Uptake, Transport, and Effects of Nano-Copper Exposure in Zucchini (Cucurbita pepo). Science of the Total Environment 665,100-106.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Tamez, C., Morelius, E., Hernandez-Viezcas, J., Peralta-Videa, J.R., Gardea-Torresdey, J. 2019. Biochemical and physiological effects of copper compounds/nanoparticles on sugarcane (Saccharum officinarum). Science of the Total Environment 649, 554-562.
  • Type: Book Chapters Status: Published Year Published: 2019 Citation: Pullagurala, V. L. R., Adisa, I.O., Rawat, S., White, J.C., Zuverza-Mena, N., Hernandez-Viezcas, J.A., Peralta-Videa, J.R., Gardea-Torresdey, J.L. (2019). Fate of engineered nanomaterials in agroenvironments and impacts on agroecosystems. In: Marmiroli, N., White, J.C., Song, J. (Eds.) Exposure to Engineered Nanomaterials in the Environment. Elsevier, chapter 4, pp. 105-142 .
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Per�z, C.D.P.; De La Torre Roche, R.; Zuverza-Mena, N.; Ma, C.; Shen, Y.; White, J.C.; Amp�lio Pozza, E.; Pozza, A.A.A.; Elmer, W.H. 2019. Metalloid and metal oxide nanoparticles suppress Sudden Death Syndrome of soybean. J. Ag. Food Chem. doi.org/10.1021/acs.jafc.9b06082.
  • Type: Book Chapters Status: Published Year Published: 2019 Citation: Tan W., Deng, C., Peralta-Videa, J., Gardea-Torresdey, J. L., 2019. Interaction of nanomaterials in secondary metabolites accumulation, photosynthesis, and nitrogen fixation in plant systems. In: Verma, S. K., Das, A. K. (Eds) Analysis, fate, and toxicity of engineered nanomaterials in plants. Comprehensive Analytical Chemistry. Elsevier, chapter 3, pp. 55-74.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Rawat, S., Adisa I. O., Wang, Y., Sun, Y., Fadil, A. S., Niu, G., Sharma, N., Hernandez-Viezcas, J. A., Peralta-Videa, J. R., Gardea-Torresdey, J. L. 2019. Differential physiological and biochemical impacts of nano vs micron Cu at two phenological growth stages in bell pepper (Capsicum annuum) plant. NanoImpact 14, 100161.


Progress 03/01/18 to 02/28/19

Outputs
Target Audience:The primary target audience for this project includes plant pathologists and growers interested in novel methods of suppressing root crop disease and enhancing overall agricultural productivity. State and federal regulators interested in nanotechnology are also a target audience for this project. Last, those concerned with the sustainable use of nanotechnology in agriculture are also a target audience for this project. Changes/Problems:The UTEP graduate student will be finishing his PhD by January 2019 (and will be joining the laboratory of the CAES PI as a post-doc). The UTEP CoPI will move another graduate student onto this project for the last year. What opportunities for training and professional development has the project provided?CAES hosted a graduate student from the Federal University of Lavros in Brazil from March 2017-March 2018. This collaboration resulted in a submitted a manuscript and since returning to Brazil, the student has become part of an active nanoparticle program on coffee diseases. A second student from this institution has arrived and will be working on this project until March 2019. In addition, three undergraduate interns from local colleges (University of New Haven, Albertus Magnus College) volunteered in CAES laboratories in 2018 to work on the NP projects. Separately, a second year of a USDA-funded research experiences for undergraduates program was concluded (CAES-Southern Ct State University) and as part of this, 10 undergraduates worked on a field study using NP to suppress fusarium wilt of chrysanthemum. Separately, the UTEP PhD student working on this project gave a presentation at Chemistry Research Day Seminar at the University of Texas at El Paso. Last, the CAES PI has recently hired the UTEP PhD student as a Post-doctoral Research Associate working on a project related to emerging contaminants in food; the UTEP graduate student will defend his PhD in the coming weeks and join the CAES in Feb-March 2019. How have the results been disseminated to communities of interest?During the current reporting period, results were presented at a number of professional meetings/venues. White (CAES PI) made presentations of project results at the University of Massachusetts Food Science Scholarship program, at Georgia Southern University (invited lecture), the University of Massachusetts Stockbridge School of Agriculture seminar series, at Jiangnan University in Wuxi China (invited lecture), at a Gordon Conference focused on nanotechnology and agriculture, at the International Conference on the Environmental Effects of Nanoparticles and Nanomaterials (Duke University), at the 15th annual International Phytotechnologies Conference (Novi Sad, Serbia), at the second Quantifying the Exposure to Engineered Nanomaterials workshop (Washington DC), at Zhejiang University in Hangzhou China (invited lecture), and at the annual Sustainable Nanotechnology Organization (SNO) Conference (Arlington VA). Elmer (CAES CoPI) made presentations of project data at the CAES seminar series (New Haven CT), at the University of Connecticut graduate student seminar series (Storrs, CT), at the annual International Integrated Pest Management symposium on "The role of Nanoparticles in IPM in Baltimore MD, at the Ag-Innovate conference in San Francisco, CA, at the Annual meeting of the American Phytopathological Society in Boston, MA, at the CAES Plant Science Day Short talk in Hamden, CT; at the Center for Sustainable Nanotechnology fall meeting in New Haven, CT and at the Second Quantifying Engineered Nanomaterials in Manufactured Products Workshop (Washington, DC). Dimkpa (IFDC CoPI) gave presentations of project data at the Materials Innovation for Sustainable Agriculture Symposium at the University of Central Florida (Orlando, FL), at the annual Agronomy Society of America Meeting (Baltimore, MD), at the annual "USA Training and Study Tour on Technology Advances'' (a program is organized annually by the International Fertilizer Development Center for foreign and local agricultural practitioners involved with fertilizers and crop nutrition), at the Myanmar Soil Fertility and Fertilizer Management Conference (Nay Pyi Taw, Myanmar), at a conference on the "Connections Between Soil Health and Human Health'' (Silver Spring, MD). Gardea-Torresdey (UTEP CoPI) presented project findings at the North China Electric Power University (Beijing, China), at the Institute of High Energy Physics, Chinese Academy of Sciences (Beijing, China), at the Gordon Research Conference on Nano-Enabled Technologies to Improve Efficiency, Quality, and Health in Food and Agriculture (Mount Holyoke College, South Hadley, MA) at the IAP2018 10th International Conference "Interfaces Against Pollution, Institut Charles Gerhardt Montpellier, Université de Montpellier, 2 place Eugène Bataillon (Montpellier, France), at the Stevens Institute of Technology (Hoboken, NJ), at the Seventh Annual Sustainable Nanotechnology Organization Conference (SNO) (Arlington VA). What do you plan to do during the next reporting period to accomplish the goals?At CAES, the ICP and electron microscopic analyses for all the samples in the above-mentioned experiments will be completed and analyzed. Manuscripts on the eggplant and pumpkin studies will be prepared for publication for peer review in journals such as Plant disease, Crop protection and/or Environmental Science: Nano. A graduate student from the Univesidad Autónoma de Querétaro in Mexico will be visiting CAES from January to July 2019; that student will conduct work on the impact of silica-based nanoparticles on pepper plants, including endpoints related to toxicity, disease and nutrition. At IFDC, a manuscript describing a study on the effect in crops of mixed exposure to metallic nanoparticles and their omissions will be written. In addition, a study on the role of ZnO nanoparticles in plant (sorghum) productivity and nutrient acquisition under drought stress will be completed. At UTEP, a manuscript focused on the impact of nanoceria on the nutritional profile of tomato fruit will be completed. Additional students focused on the impact of nanoceria on different cultivars of tomato and potentially other crops will be initiated.

Impacts
What was accomplished under these goals? At CAES, work was completed and a manuscript was submitted that assessed the role of metal oxides and metalloid nanoparticles of Ag, B, Cu, Ce, Mn, and Zn on the suppression of Soybean Sudden death syndrome. NP of CuO and ZnO were the most disease suppressive in four greenhouse experiments. Digestion of root and above ground tissue found NP application increased the respective elements but did not affect the uptake of other elements. A third year of field studies examining the individual and combined effects of NP CuO, MnO, and ZnO on the yield of eggplant was completed. When data from all years were combined, no differences were observed in total bulk yield, but trends each year suggested the combination treatments of CuO, MnO and ZnO or MnO and ZnO provided the greatest yield. Further analysis of yield components (early vs late yield, marketable vs total yield) are underway. A two year pumpkin field study at two sites comparing NP of CuO, MnO, SiO2, and ZnO to their respective salts for control of powdery mildew was also completed. Treatments that consistently reduced the disease severity indices in Hamden both years and in Griswold in 2017 were CuO NPs and the conventional fungicide sprays. No significant difference was observed in Griswold in 2018, but the trend was for plots treated with CuO NPs to have the lowest disease severity. Although none of the treatments resulted in statistically significant increases in yield, NPs CuO tended to have the highest yield values. At UTEP, a follow up study on a previously published paper (Adisa et al., 2018) was conducted. The impact of the cerium compounds (nano-CeO2 and cerium acetate) on the nutritional value of tomato fruit, grown in Fusarium oxysporum infested soil, was investigated. Fruit biomass, water content, size, and nutritional properties (lycopene, reducing and total sugar, cerium, micro- and macro-elements) were evaluated. Fusarium infection decreased lycopene in the fruit by 17%, compared to the control. Cerium treatments did not alter the lycopene content in both infested and non-infested fruits. Exposure of nCeO2 at 250 mg/kg, via roots, increased Ca by 261% in fruit of infested plants, compared with non-infested nCeO2 treated plants (p ≤ 0.05). CeAc, at 50 mg/L, decreased fruit biomass by 22% in infested tomato plants (p ≤ 0.05). Overall, the findings indicate that NP CeO2 has potential to suppressed Fusarium wilt disease and has negligible effects on the nutritional value of tomato fruit (article in preparation). Currently, another experiment is underway to examine the morphological (microscopy), physiological and biochemical impacts, of root and foliar applications of copper compounds (nCuO and CuSO4) at 250 and 500 mg/L , and commercial fungicide, on Fusarium infested tomato plants. Using a confocal microscope, the reactive oxygen species (ROS) in the root tissues will be measured and correlated with the enzyme activities, as well as the pathogenesis-related gene expression. SEM will also be employed to view any morphological changes in the root architecture. At IFDC, investigations on Zn and Mn effects on crop use of NPK initiated in a previous reporting period was completed, and manuscript preparation was initiated. Separately, an addition-omission strategy was used to address particle-size and nutrient-specific effects in soybean exposed to a mixture of nano or bulk oxide particles of Zn (2 mg Zn/kg), Cu (1 mg Cu/kg) and B (1 mg B/kg) in soil. The mixture of oxide particles of both sizes significantly promoted grain yield and Zn accumulation, but suppressed P accumulation. However, exposure to the nano-oxides specifically stimulated shoot growth (47%), flower development (63%), shoot biomass (34%), and N (53%) and K (42%) accumulation. Omission of individual nutrients from the mixtures evoked significant responses that were nano or bulk-specific, including promotion of shoot growth by bulk-B; inhibition of flower development by nano-Cu; stimulation of flower development by bulk-B; suppression of grain yield by nano-Zn; enhancement of Cu uptake by nano-B; enhancement of B uptake by bulk-Cu; stimulation of P uptake by nano-Zn or bulk-B; enhancement of residual soil N and Zn levels by nano-Cu; and enhancement of residual soil Cu level by nano-Zn and nano-B. Collectively, Zn was largely responsible for driving the agronomic (biomass and grain yield) responses in this soil, with concurrent ramifications in environmental management (N and P) and human health (Zn nutrition). Overall, compared to bulk nutrients, nano-scale nutrients played a greater role in evoking plant responses. Last, the IFDC CoPI visited CAES laboratories for several days during the current reporting period. Last, we note that this project (with the help of the USDA NIFA NPL, Dr. Hongda Chen) was featured in the FY 2019 National Nanotechnology Initiative (NNI) Supplement to the President's 2019 Budget. This also led directly to a meeting with the White House Office of Science and Technology Policy (OSTP) staff at the Eisenhouer Executive Office Building where project results were discussed.

Publications

  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Elmer, W.; De La Torre-Roche, R.; Pagano, L.; Majumdar, S.; Zuverza-Mena, N.; Dimkpa, C.; Gardea-Torresdey, J.; White, J.C. 2018. Effect of foliarly applied nanoparticle CuO on Fusarium wilt of watermelon. Plant Disease 102:1394-1401.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Liao, Y.Y.; Strayer-Scherer, A.; White, J.C.; Mukherjee, A.; De La Torre-Roche, R.; Ritchie, L.; Clark, D.; Vallad G. E.; Freeman, J.; Jones, J. B; Paret, M.L. Nano-Magnesium oxide: A novel bactericide against copper-tolerant Xanthomonas perforans causing tomato bacterial spot. Phytopathol. J. 108 (10), 146-147.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adisa, I.; Pullagurala, V.L.R.; Rawat, S.; Hernandez-Viezcas, J.A.; Dimkpa, C.O.; Elmer, W.H.; White, J.C.; Peralta-Videa, J.R.; Gardea-Torresdey, J.L. 2018. Role of cerium compounds in Fusarium wilt suppression and growth enhancement in tomato (Solanum lycopersicum). J. Agric. Food Chem. 66:5959-5970.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dimkpa, C.O.; Adisa, I.O.; Elmer, W.E.; Gardea-Torresdey, J.L.; White, J.C. 2018. Size-specific effects of nanoparticle Mn exposure on nutrient acquisition and distribution in wheat. Agronomy 8(9), 158; doi.org/10.3390/agronomy8090158.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dimkpa, C.O.; Singh, U.; Bindraban, P.E.; Elmer, W.E.; Gardea-Torresdey, J.; White, J.C. 2018. Exposure to weathered and fresh nanoparticle and ionic Zn in soil promotes grain yield and modulates nutrient acquisition in winter wheat (Triticum aestivum). J.Agric. Food Chem. 66 (37), 9645-9656.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Guo, H.; White, J.C.; Wang, Z.; Xing, B. 2018. Nano-enabled fertilizers to control the release and use efficiency of nutrients. Curr. Op. Environ. Sci. Health doi.org/10.1016/j.coesh.2018.07.009.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Elmer, W.H.; Ma, C.; White, J.C. 2018. Nanoparticles for plant disease management. Curr. Op. Environ. Sci. Health 6:66-70.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Borgatta, J.; Ma, C.; Hudson-Smith, N.; Elmer, W.; Plaza Pére, C.D.; De La Torre-Roche, R.; Zuverza-Mena, N.; Haynes, C.L.; White, J.C.; Hamers, R.J. 2018. Copper nanomaterials suppress root fungal disease in watermelon (Citrullus lanatus): Role of particle morphology, composition and dissolution behavior. ACS Sustain. Chem. Eng. 6 (11), pp 14847⿿14856.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pullagurala, V.L.R., Adisa, I O., Rawat, S., Kalagara, S., Hernandez-Viezcas, J.A., Peralta-Videa, J.R., Gardea-Torresdey, J.L. (2018). ZnO nanoparticles increase photosynthetic pigments and decrease lipid peroxidation in soil grown cilantro (Coriandrum sativum). Plant Physiology and Biochemistry 132, 120⿿127.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pullagurala, V.L.R., Adisa, I.O., Rawat, S., Kim, B, Barrios, A.C., Medina-Velo, I.A., Hernandez-Viezcas, J.A., Peralta-Videa, J.R., Gardea-Torresdey, J.L. (2018). Finding the conditions for the beneficial use of ZnO nanoparticles towards plants-A review. Environmental Pollution 241, 1175-1181.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Medina-Velo, I. A., Zuverza-Mena, N., Tamez, C., Ye, Y., Hernandez-Viezcas, J. A., White, J.C., Peralta-Videa, J. R., Gardea-Torresdey, J.L. (2018). Minimal transgenerational effect of ZnO nanomaterials on the physiology and nutrient profile of Phaseolus vulgaris. ACS Sustainable Chemistry & Engineering 6 (6), 7924⿿7930.


Progress 03/01/17 to 02/28/18

Outputs
Target Audience:The primary target audience for this project includes plant pathologists and growers interested in novel methods of suppressing root crop disease and enhancing overall agricultural productivity. State and federal regulators interested in nanotechnology are also a target audience for this project. Last, those concerned with the sustainable use of nanotechnology in agriculture are also a target audience for this project. Changes/Problems:In August of this year, PI J.C. White was awarded a seed grant from the Center for Sustainable Nanotechnology (CSN). The CSN is funded by the NSF Center forChemical Innovation program at approximately20 million dollars over 5 years and is focused on the synthesis and characterization of novel advanced materials. The Center leadership decided to fund a post-doctoral researcher at CAESfor two years (approximately 125,000 dollars per year) to explore the new avenue of disease suppression through the application of novel nanoscale materials they synthesize. The work as part of the CSN will run directly parallel to the current USDA project and the additional resources afforded by CSN should significantly advance the USDA project goals and products. What opportunities for training and professional development has the project provided?The UTEP graduate student and CAES post-doc each gave platform presentions at the 6th Annual Sustainable Nanotechnology Organization conference in Los Angeles CA. The IFDC CoPI also gave a presentation at the USA Training and Study Tour on Technology Advances that was organized by IFDC and focused on the application of nanotechnology in the fertilizer industry. How have the results been disseminated to communities of interest?During the current reporting period, results from this project were presented at a number of professional meetings/venues, including at the University of Connecticut Department of Pathobiology and Veterinary Science Seminar Series, a WebEx to the 11 universities that are part of the NSF-funded "Center for Sustainable Nanotechnology, at the University of Nebraska Water Center seminar series, a WebEx to the Florida Department of Agriculture and Consumer Services "Ag Science Café", at the University of Connecticut Department of Nutritional Sciences seminar series, at the 253rd American Chemical Society National Meeting in San Francisco CA, at the Harvard School of Public Health, at the International Symposium on Emerging Contaminants and Environmental Nanotechnology which was held at Nankai University in Tianjin China, at the 3rd Annual Parma Nano-Day at the University of Parma in Parma Italy, at the 2017 International Conference on the Biogeochemistry of Trace Elements (ICOBTE) in Zurich Switzerland, at Zhejiang University in Hangzhou China as part of a new 111 Project entitled "Crop Produce Quality and Safety" which was awarded this summer from the Chinese Ministry of Education and the State Administration of Foreign Experts Affairs, at the 14th International Phytotechnologies Conference in Montreal Canada, as a Brace Seminar at McGill University in Montreal Canada, at the AgroBio Nanotechnology Conference in Saltillo Mexico which was organized for the Center for Applied Analytical Chemistry (CIQA), at the 6th annual Sustainable Nanotechnology Organization (SNO) Conference in Los Angeles CA, at the Materials Research Society (MRS) in Boston MA, at the Bazilian Congress of Soil Science in Belem City Brazil, and at Nanjing Agricultural University in Nanjing China. In addition, data on this project was presented at the annual Connecticut Agricutlural Experiment Station open house event (Plant Science Day). Last, three peer reviewed manuscripts were published. What do you plan to do during the next reporting period to accomplish the goals?In the upcoming reporting period, at IFDC the elemental and microscopic analysis for the Zn and Mn nanofertilizer experiments will be completed. At all institutions, greenhouse and field investigations will be initiated to evaluate the effect of multiple applications of nanoscale nutrients on disease suppression and plant growth. A number of plant-disease-nanoparticle combinations will be explored. In addition, a molecular-based study will be initiated at CAES with sobyean and copper nanomaterials to evaluate the mechanistic nature of crop response. Investigations will also begin to evaluate composite and multi-nutrient nanoscale materials (i.e.; chitosan coated Zn-Cu nanoparticle). At UTEP, the potential of different forms of ceria to suppress tomato fungal pathogens and enhance yield will continue. Last, the use of scanning/transmission electron microscopy with energy dispersive X-ray spectroscopy (S/TEM-EDX) as an endpoint for various investigations will begin in earnest.

Impacts
What was accomplished under these goals? At CAES, themanuscript focused on the control of Fusarium in water melon by foliar nanoscale micronutrient amendments was submitted for publication and is now in press. A series of field and greenhouse trials focusing on the use of various new nanoscale materials (some from the Center for Sustainable Nanotechnology) to control fusarium was also initiated and completed; biomass, disease data, pigment production and other physiological data is currently being analyzed and digestion/ICP analsyis of the tissues for nutritional content is ongoing. A series of field trials looking at root fungal pathogen control in strawberry, as well as foliar pathogen control in pumpkins and grapes were conducted. In all cases, data analysis is ongoing. At UTEP,a new batch of experiments was set up to examine the role of cerium compounds (nano-CeO2 and cerium acetate) in the suppression of fusarium wilt disease in Bonny Best tomato cultivar. The plants were cultivated until full maturity (126 days). Disease severity, fruit production, biochemical and physiological parameters were analyzed. Root and foliar applications of nano-CeO2 at 250 mg/L reduced the disease severity by 53 % and 57%, respectively, compared with the infested control. Similarly, there was a significant reduction in the spread of the disease in plants exposed to CeAc at 250 via roots (35%) or foliage (41 %), compared with the infested controls. Foliar exposure of nano-CeO2 at 250 mg/L increased the chlorophyll content in infested plants by 28 %. This treatment also increased fruit weight of infested and non-infested plants by 37% and 60 %, compared with the respective controls. Overall, the data show that nano-CeO2 has potential to control fusarium wilt and increase production in tomato plants. This data is currently being drafted into a manuscript. At IFDC, a study is evaluating the separate responses of wheat to zinc (Zn) and manganese (Mn) micronutrient fertilization as part of efforts to further our understanding of how micronutrients influence use of NPK by crops. The micronutrients were applied as ZnO nanopowder (NP) or Zn-sulfate (salt), and MnO NP, bulk MnO powder, and Mn-chloride (salt). The rate of Zn used was 6 mg/kg soil, and that of Mn was 10 mg/kg soil. N, P, and K rates were 200, 75 and 200 mg/kg soil, respectively. In the Zn study, the experiment was conducted using fresh and used (previously treated with the same Zn types and amount, and cropped with sorghum) soils. This later treatment will demonstrate whether Zn as NP or salts (ions) has any residual value as fertilizer for subsequent crops, compared to fresh Zn applications. Compared to the respective control (NPK only), Zn treatment as NP or salts (ions) increased grain yield by 15% or 19%, respectively, in the fresh soil. Wheat grain yield was similarly increased in the old soil, by 10.5% and 12.5% respectively by the residual NP and ionic Zn. These findings indicate that Zn treatment has both immediate and residual effects on crop production. However, a greater effect was observed in the used soil compared to the fresh soil, with a 33% and 31% grain yield increases by NP and ionic Zn, respectively in the used soil. Also, ionic Zn caused greater yield enhancing effect than NP Zn in both soil types. Also, compared to the control (NPK only), Mn treatment in soil as NPs, salts (ions), or bulk Mn increased grain yield by 16%, 9% and 12%, respectively. Mn NPs resulted in 4.6% more grain yield when applied as a foliar treatment, compared to soil application. These findings indicate that NPs of Mn are more effective than other Mn forms in enhancing wheat grain yield. Also, compared to soil application, foliar application of NPs of Mn may be more effective at increasing grain yield. Elemental and electron microscopic analysis of various tissues from these trials is currently being undertaken.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Dimkpa, C; White, J.C.; Elmer, W.; Gardea-Torresdey, J. 2017. Nanoparticle and ionic Zn promote nutrient loading of sorghum grain under low NPK fertilization. J. Agric. Food Chem. 65:85528559.
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Elmer, W.; De La Torre-Roche, R.; Pagano, L.; Majumdar, S.; Zuverza-Mena, N.; Dimkpa, C.; Gardea-Torresdey, J.; White, J.C. Effect of foliarly applied nanoparticle CuO on Fusarium wilt of watermelon. Plant Disease In press.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Dimkpa C., Bindraban P. 2017. Nanofertilizers: new products for the industry? 2017. Journal of Agricultural and Food Chemistry, DOI: 10.1021/acs.jafc.7b02150


Progress 03/01/16 to 02/28/17

Outputs
Target Audience:The primary target audience for this project includes plant pathologists and growers interested in novel methods of suppressing root crop disease andenhancing overall agricultural productivity. State and federal regulators interested in nanotechnology are also a target audience for this project. Last,those concerned with the sustainable use of nanotechnology in agriculture are also a target audience for this project. Changes/Problems:One of the CoPIs on this project, C. Dimpka, hasmoved from the VFRC in Washington DC to the parent organization, IFDC, which is in Muscle Shoals Alabama. This move has allowed the investigator more direct and continual access to greenhouse facilities (the original plan was remote supervision), which will increase project output. Second, sorghum has been added as a crop to replace corn; this new crop better aligns with IFDC goals and related projects. At UTEP, additional investigations looking at the effect of cultivar type on treatment effects have been added. This was in part based on initial data showing the first selected cultivar was highly resistant to infection. Last, at CAES transcriptomics has been added as another parameter to measure plant response to infection and nanoscale nutrient amendment. What opportunities for training and professional development has the project provided?A CAES Post-doctoral Associate was sent to the 5th annual meeting of the Sustainable Nanotechnology Organization and gave a platform presentation entitled "Nanoscale micronutrients suppress plant disease and increase crop yield." How have the results been disseminated to communities of interest?Results were presented in both platform and poster form at4 professional scientific meetings. One of these venues was an international conference hosted by the Chinese Academy of Agricultural Sciences and USDA that focused on sustainable nanotechnology in agricutlure. In addition, CAES project results (including an ongoing field demonstration) were presented at the agency annual open house (Plant Science Day) held in August of 2016; the event was open to the public and over 1100 attendees were present. The CAES investigators spoke to threeseparate reporters during the current period; the linksare: https://www.chemistryworld.com/news/metal-micronutrients-get-to-the-root-of-antifungal-defence/1017334.article http://www.ozy.com/fast-forward/could-nanotechnology-end-hunger/70771 http://www.nhregister.com/health/20161226/connecticut-scientists-studying-nanoparticles-to-help-grow-disease-resistant-food Last, we have created a website (http://www.ct.gov/caes/cwp/view.asp?a=4898&q=585400 ) for our USDA NIFA AFRI Center of Excellence entitled The Center for Nanotechnology and Agricultural Pathogens Suppression (CeNAPS). What do you plan to do during the next reporting period to accomplish the goals?Ongoing greenhouse trials described above at CAES, UTEP, and IFDC will be completed, including extensive analysis by ICP-MS for element content in various tissues and by electron microscopy on specific tissues to detect the presence of nanoparticle forms of amended nutrients. The CAES eggplant trials described above will be repeated in 2017 to confirm findings. At UTEP, additionalcultivars of tomato will be investigated under the current protocols. Last, preliminary studies assessing the impacts of soil type and NP source will be planned. A manuscript describing the CAES greenhouse and field work with watermelon will be prepared for submission to a peer reviewed journal, as will the IFDC project on micronutrients and drought stress.

Impacts
What was accomplished under these goals? CAES: At CAES, experiments investigating eggplant and tomato were initiated during the current period. First, a series of greenhouse and field experiments were established to assess combinations of nanoparticles (NP) of CeO2, CuO, MnO, and ZnO on eggplant growth upon infection with Verticillium wilt. In an analysis of preliminary data, all treatments performed better than control in total yield, and combinations of Mn and Zn performed the best. Tissue samples of fruit, roots, stems, and leaves are currently being processed for elemental composition by ICP-MS. Second, a series of greenhouse experiments with watermelon were conducted to compare NP CuO to other products. Two greenhouse studies showed that NP CuO were superior to NP B, CeO, MnO, TiO, SiO or ZnO in suppressing Fusarium wilt on Watermelon and for promoting growth. ICP-MS showed twice as much Cu in the roots of plants sprayed with NP of CuO than controls or other treatments. Three field trials have been completed. A two year study conducted in Hamden, CT in 2015 and 2016 compared NP CuO to bulk CuO, Kocide 2000 (a CuOH-containing fungicide), Cu octanoate (an organic Cu fungicide) and an untreated control. Although yield differences were not statistically significant, in both years seedling transplants sprayed with NP CuO produced the highest yield and had lowest disease ratings. Fruit digests from 2015 field studies found no differences in elemental content of the edible tissue. Another field study conducted in Griswold, CT compared NP B, CeO, CuO, MnO, or ZnO to controls in Fusarium-infested soil or in noninfested soil. As before, plants treated with NP of CuO produced the highest yield in infested soil, but significant differences were not observed. Transcriptomic analysis was performed on roots of watermelons treated with Cu, Fusarium, the combination, and then were compared to controls in three separate experiments. Significant upregulation of polyphenol oxidase genes (a Cu activated enzyme associated with host defense) and of the Plant Resistance 1 protein (associated with resistance) were consistently observed when NP CuO and Fusarium were combined. We hypothesize that NP CuO may activate defense mechanisms in plants, likely via basipetal translocation of the nanoscale nutrient. UTEP: At UTEP, an experiment was set up in the greenhouse in mid July 2016 to examine the efficacy of root and foliar application of ZnO nanoparticle and ZnSO4 to control fusarium wilt disease on tomato. Plants were exposed to the Zn compounds at the concentrations previously specified in the protocol. Three week old seedlings were transplanted to soil and allowed to grow for one week before inoculation with the pathogen Fusarium oxysporum. Plants grew for 72 days without showing symptoms of infection. Roots were examined to determine the disease severity but no visual presence of fungal hyphae in the roots was found. This was due to either a small amount of inoculum or the strong resistance of the cultivar Tiny Tim Cherry Tomato used for the experiment. A second greenhouse experiment was set up early November 2016 using the tomato variety Bush Beefsteak, which is more susceptible to Fusarium infection. Plants were transplanted in late November and were inoculated with high amount of pathogen. Different variables such as disease severity, photosynthetic parameters, production, and fruit quality are currently being collected until full maturity. IFDC: At IFDC, a greenhouse study was established in July 2016 to evaluate the effect of ZnO nanoparticles versus Zn salts on the growth, yield and NPK use efficiency of sorghum as a function of soil versus foliar application. The plants have recently been harvested and data is now being collated and assessed for different parameters. Samples for microscopic analysis to determine presence or absence of ZnO NPs in shoot were sent to CAES for imaging. ICP-MS is ongoing for nutrient content. Second, a winter wheat crop was established during late November 2016 to evaluate the effect of residual ZnO nanoparticles (and Zn salt) from prior soil application versus new ZnO application on growth, yield, NPK use and Zn content enhancement in grain. Third, another winter wheat crop was established during late November 2016 to evaluate the effect on growth, yield and NPK use efficiency of NP Mn2O3 versus Mn salt applied as either a soil or foliar treatment. Last, a project conducted in conjunction with US AID and current grant funds was completed. The study was on the effect of composite NP micronutrients (ZnO, CuO and B2O3) versus salts of these elements on soybean vegetative and reproductive development, as well as NPK use, as a function of drought stress.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Dimkpa C., Bindraban P., Fugice J., Agyin-Birikorang S., Singh U., Hellums D. 2017. Composite micronutrient nanoparticles and salts decrease drought stress in soybean. Agronomy for Sustainable Development. DOI: 10.1007/s13593-016-0412-8.