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Phyto-toxicological Effects of Copper Nanoparticles in Bell Pepper ( Capsicum annum ) plants Swati Rawat ESE PhD Student Gardea Group, University of Texas at El Paso Sustainable Nanotechnology Organization Orlando, FL, November, 2016 1

Structure of the presentation  Introduction - Nanoparticles - Bell pepper plants  Methodology  Results  Conclusions 2

Introduction 3

Factors affecting NPs induced toxicity towards terrestrial plants Size nZnO Surface charge nCeO 2 H nSi Ca 2+ C=O MWCNT’s COO Mg 2+ nCuO P H H + OH - Structure and defects Reddy, P. V. L., Hernandez-Viezcas, J. A., Peralta-Videa, J. R., & Gardea-Torresdey, J. L. (2016). Lessons learned: Are engineered nanomaterials toxic to terrestrial plants?. Science of The Total Environment , 568 , 470-479.

Copper Nanoparticles (NPs) SEM Micrographs of copper nanoparticles Hong, Jie, Cyren M. Rico, Lijuan Zhao, Adeyemi S. Adeleye, Arturo A. Keller, Jose R. Peralta-Videa, and Jorge L. Gardea-Torresdey. "Toxic Effects of Copper-Based Nanoparticles Or Compounds to 5 Lettuce (Lactuca Sativa) and Alfalfa (Medicago Sativa)." Environmental Science: Processes & Impacts 17, no. 1 (2015): 177-185.

Applications of Copper NPs 6

Global flows for Cu and oxides of Cu (metric tons/yr) in 2010 Keller, Arturo A., Suzanne McFerran, Anastasiya Lazareva, and Sangwon Suh. "Global Life Cycle Releases of Engineered Nanomaterials." Journal of 7 Nanoparticle Research 15, no. 6 (2013): 1-17.

Bell pepper plants Capsicum annum  Rich in anti-oxidants like carotenoid, sugars, vitamin C.  Fruit is 92% water, rest are carbohydrates and small amount of protein and fat https://authoritynutrition.com/foods/bell-peppers 8

Methodology 9

Soil  Soil collected on the east side of El Paso, TX.  Soil characterization conducted on Malvern Mastersizer Hybrid 2000G - Sand : 19.7 % - Silt : 64.92 % - Clay : 15.38 %  Natural soil : silt loam 10

Sowing seeds at the green house for seedling transplantation 11

Preparing pots in the lab 12

Plant growth stages : full growth cycle 90 days Seedlings growing Plants 10 days post transplantation Seedlings ready for Freshly transplanted transplantation seedlings 13

Plant growth stages : full growth cycle 90 days Plants 30 days Fully matured plants, post 90 days post transplantation transplantation Plants 60 days post Plants 45 days post transplantation, fruiting transplantation, flowering 14

Conditions at the green house  Controlled environment, temperature, relative humidity, and light intensity - Average light 10.1 mol/m 2 /d - Average day temperature 27.2± 1.6˚C - Average night temperature 25± 2.1˚C  Water every other day, or as need be with fertilizer solution, 15-5-15 ratio of N-P 2 O 5 -K 2 O, pH: 5.8, EC: 1.00 mS/cm  Abamectin, Avid 0.15 EC , to treat aphids or white fly 15

Harvesting 16

Gas exchange measurement: LI-6400XT portable photosynthesis system 17

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Acid digestion and sample analysis on the ICP-OES 19

Results 20

Chlorophyll content, nCuO vs ionic copper treatments Chlo lorophyll ll Con ontent 80 Relative Chlorophyll Content (SPAD) 70 60 50 nCuO 40 CuCl2 30 20 10 0 Control 62.5 125 250 500 Concentration of the treatments, mg/kg 21

Gas Exchange : Evapotranspiration, nCuO vs ionic copper treatments Evapotranspiration 6 b ab Evapotranspiration mmol/m 2 /s 5 ab ab a 4 3 nCuO a a CuCl2 a ab 2 1 0 Control 62.5 125 250 500 Concentration of the treatments, mg/kg 22

Gas Exchange: Stomatal conductance, nCuO vs ionic copper treatments St Stomatal l Con onductance 500 450 c bc Stomatal Conductance mol/m2/s 400 abc 350 abc abc 300 250 nCuO 200 CuCl2 150 a ab 100 a a 50 0 Control 62.5 125 250 500 Concentration of the treatments, mg/kg 23

Gas exchange : Photosynthesis, nCuO vs ionic copper treatments Photosynthesis 18 b ab 16 ab ab 14 Photosynthesis µmol/m 2 /s ab 12 10 nCuO ab a 8 a ab CuCl2 6 4 2 0 Control 62.5 125 250 500 Concentration of the treatments, mg/kg 24

Elemental analysis of root samples, copper 350 c 300 mg of Cu/kg of root dry wt . 250 200 b b b b 150 ab ab ab 100 a a a a 50 0 Control 62.5 62.5 Control 125 125 Control 250 250 Control 500 500 nCuO CuCl2 nCuO CuCl2 nCuO CuCl2 nCuO CuCl2 Treatment Concentration, mg/kg 25

Elemental analysis of leaves samples, copper 30 c bc 25 ab bc bc mg of Cu/kg leaves dry wt. abc abc 20 abc a 15 a a a 10 5 0 Control 62.5 62.5 Control 125 125 Control 250 250 Control 500 500 nCuO CuCl2 nCuO CuCl2 nCuO CuCl2 nCuO CuCl2 Treatment Concentration, mg/kg 26

Elemental analysis of fruit samples, copper 14 a a a 12 a a a a a 10 a a a a mg of Cu/kg of fruit dry wt. 8 6 4 2 0 Control 62.5 62.5 Control 125 125 Control 250 250 Control 500 500 nCuO CuCl2 nCuO CuCl2 nCuO CuCl2 nCuO CuCl2 Treatment Concentration, mg/kg 27

Conclusions  Gas exchange : evapotranspiration, stomatal conductance, and photosynthesis were not significantly different with respect to the control but were statistically different with respect to each other at the different concentrations of nCuO and CuCl 2 .  The copper content in root samples was significantly increased at 125 mg/kg CuCl 2 , 250 mg/kg nCuO and CuCl 2, andat 500 mg/kg nCuO and CuCl 2 wrt the control. The two treatments were significantly different at the highest concentration.  The leaf samples found significantly higher amount of copper at 250 mg/kg and 500 mg/kg concentration of both the compounds wrt the control.  Significantly higher amount of copper was found in the fruit samples at 125 mg/kg ionic treatment. 28

Acknowledgements  UCCEIN for funding the research  Texas AnM Agrilife Research and Extension Centre at El Paso, TX.  University of Texas at El Paso  Lab Mates  Faculty - Dr Youping Sun - Dr Jose A. Hernandez - Dr Jose R. Peralta - Dr Jorge Gardea Torresday  The SNO conference organizers 29

References  Reddy, P. Venkata Laxma, J. A. Hernandez-Viezcas, J. R. Peralta-Videa, and J. L. Gardea-Torresdey. "Lessons Learned: Are Engineered Nanomaterials Toxic to Terrestrial Plants?" Science of the Total Environment 568, (10/15, 2016): 470-479.  Hong, Jie, Cyren M. Rico, Lijuan Zhao, Adeyemi S. Adeleye, Arturo A. Keller, Jose R. Peralta-Videa, and Jorge L. Gardea- Torresdey. "Toxic Effects of Copper-Based Nanoparticles Or Compounds to Lettuce (Lactuca Sativa) and Alfalfa (Medicago Sativa)." Environmental Science: Processes & Impacts 17, no. 1 (2015): 177-185.  Keller, Arturo A., Suzanne McFerran, Anastasiya Lazareva, and Sangwon Suh. "Global Life Cycle Releases of Engineered Nanomaterials." Journal of Nanoparticle Research 15, no. 6 (2013): 1-17. 30

Thank You! Questions ? 31