Abstract
Widespread application of silver nanoparticles (AgNPs), due to their antibacterial and antifungal properties, increases their release into the environment and potential detrimental impact on living organisms. Plants may serve as a potential pathway for AgNPs bioaccumulation and a route into the food chain, hence investigation of AgNP phytotoxic effects are of particular importance. Since proteins are directly involved in stress response, studies of their abundance changes can help elucidate the mechanism of the AgNP-mediated phytotoxicity. In this study, we investigated proteomic changes in tobacco (Nicotiana tabacum) exposed to AgNPs and ionic silver (AgNO3). A high overlap of differently abundant proteins was found in root after exposure to both treatments, while in leaf, almost a half of the proteins exhibited different abundance level between treatments, indicating tissue-specific responses. Majority of the identified proteins were down-regulated in both tissues after exposure to either AgNPs or AgNO3; in roots, the most affected proteins were those involved in response to abiotic and biotic stimuli and oxidative stress, while in leaf, both treatments had the most prominent effect on photosynthesis-related proteins. However, since AgNPs induced higher suppression of protein abundance than AgNO3, we conclude that AgNP effects can, at least partially, be attributed to nanoparticle form.
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Abbreviations
- ADH:
-
alcohol dehydrogenase
- AgNPs:
-
silver nanoparticles
- ATP:
-
adenosine triphosphate
- CBB:
-
Commassie Brilliant Blue
- CBP:
-
cap-binding protein
- CHAPS:
-
3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate
- CHCA:
-
alpha-cyano-4-hydroxycinnamic acid
- Cu/Zn-SOD:
-
copper/zinc-dependent superoxide dismutase
- DTT:
-
dithiothreitol
- EF Tu:
-
elongation factor Tu
- EDTA:
-
ethylenediaminetetraacetic acid
- EDX:
-
electron dispersive X-ray
- Fd:
-
ferredoxin
- Fe-SOD:
-
iron-dependent superoxide dismutase
- FNR:
-
ferredoxin-NADP reductase
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- GDH:
-
glutamate dehydrogenase
- GLP:
-
germin-like protein
- GO:
-
gene ontology
- GS:
-
glutamine synthetase
- GST:
-
glutathione S-transferase
- ICP-MS:
-
inductively coupled plasma mass spectrometry
- IDH:
-
isocitrate dehydrogenase
- IEF:
-
isoelectric focusing
- IPG strips:
-
immobilized pH gradient strips
- KCl:
-
potassium chloride
- LOQ:
-
limit of quantification
- MALDI-TOF/TOF:
-
matrix-assisted laser desorption/ionization–time-of-flight/time-of-flight
- MDH:
-
malate dehydrogenase
- Mn-SOD:
-
manganese-dependent superoxide dismutase
- NADP+ :
-
nicotinamide adenine dinucleotide phosphate
- NADPH:
-
reduced form of NADP+
- NCBIprot:
-
National Center for Biotechnology Information protein database
- NDPK1:
-
nucleoside diphosphate kinase
- PCA:
-
principal component analysis
- PMSF:
-
phenylmethylsulfonyl fluoride
- PPIase:
-
peptidyl-prolyl cis-trans isomerase
- PR proteins:
-
pathogenesis-related proteins
- PSI:
-
photosystem I
- PSII:
-
photosystem II
- RbcL:
-
Rubisco large subunit
- ROS:
-
reactive oxygen species
- RPE:
-
ribulose-phosphate 3-epimerase
- Rubisco:
-
ribulose-1,5-bisphosphate carboxylase/oxygenase
- SDS-PAGE:
-
sodium dodecyl sulphate-polyacrylamide gel electrophoresis
- TCA cycle:
-
tricarboxylic acid cycle
- 2-DE:
-
two-dimensional electrophoresis
- TEM:
-
transmission electron microscope
- TFA:
-
trifluoroacetic acid
- TPI:
-
triose phosphate isomerase
- UDP:
-
uridine diphosphate
- UniProt:
-
Universal Protein Resource
- UTP:
-
uridine triphosphate
- VCaB42:
-
vacuole-associated annexin protein 1
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Acknowledgments
Tobacco seeds were provided from the Zagreb Tobacco Institute. Authors wish to thank Mr. Juraj Balen for technical assistance in graphical design of figures.
Funding
Performed analyses were supported by the Croatian Science Foundation [grant number IP-2014-09-6488] and the University of Zagreb [grant number 20283115]. TEM analyses have received funding from the European Union Seventh Framework Programme under Grant Agreement 312483 - ESTEEM2 (Integrated Infrastructure Initiative - I3).
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Fig. S1
Silver nanoparticles coated with citrate (AgNP-citrate) (a) TEM image, (b) bright field image, (c) energy-dispersive X-ray spectrum (PDF 305 kb)
Fig. S2
(a) Absorbance spectra of 100 μM AgNP solution in Milli-Q® water during the period of 7 days illustrating the AgNP stability during the exposure period. (b) Physico-chemical characteristics of 100 μM AgNP solution in Milli-Q® water after 0 min and 2, 5 and 7 days by means of hydrodynamic diameter (dH), ζ potential values, position of SPR peak and percentage of ionic Ag+ (PDF 111 kb)
Fig. S3
AgNP localization in the root cells after exposure of tobacco plants to 100 μM AgNPs. (a) TEM image of AgNP, (b) bright field images, and (c) energy-dispersive X-ray spectrum (PDF 177 kb)
Fig. S4.
Principal component analysis (PCA) of protein abundance values of roots (a) and leaves (b) of tobacco plants exposed to either 100 μM AgNPs or 100 μM AgNO3. The first two components accounted for 99.2% and 99% of the variability in roots and leaves, respectively. Ac – cytosol-associated annexin; AMT – aminomethyltransferase; Av – vacuole-associated annexin; ATPsyn – ATP synthase; BG – beta glucanase; CA – carbonic anhydrase; CAP – cysteine rich secretory protein; CAT – catalase; CBP20 – cap-binding protein 20; chla/b 8 – chlorophyll a-b–binding protein 8; Do-like – protease Do-like; EF Tu – elongation factor Tu; EI – elicitor inducible; ENO – enolase; Fe-SOD – iron-dependent superoxide dismutase; FNR – ferredoxin-NADP reductase; GAPDH – glyceraldehyde-3-phosphate dehydrogenase; GDH – glutamate dehydrogenase; GH – glycosyl hydrolase; GLP – germin-like protein; GRP – glycine-rich RNA-binding protein; GS – glutamine synthetase; GST – gluthatione-S transferase; HBP1 – ankyrin-repeat protein HBP1; IDH – isocitrate dehydrogenase; MDH – malate dehydrogenase; MDHARc – cytosolic monodehydroascorbate reductase; MDHARp – plastid monodehydroascorbate reductase; Mn-SOD – manganese-dependent superoxide dismutase; mRBP – mRNA binding protein; NDPK1 – nucleoside diphosphate kinase 1; Omp-A – outer membrane protein A-like; PA – plastidic aldolase; PPIase – peptidyl-prolyl cis-trans isomerase; PPX – peroxidase; PSI – photosystem I proteins; PSII – photosystem II proteins; QR – quinone reductase; RbcAc – Rubisco activase; RbcL – Rubisco large subunit; RbcL-part – partial Rubisco large subunit; RHRE – rhicadhesin GLP-like protein; RPE – ribulose-phosphate 3-epimerase; TPI – triose phosphate isomerase; UDP-GluDH – UDP-glucose dehydrogenase (PDF 15 kb)
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Peharec Štefanić, P., Jarnević, M., Cvjetko, P. et al. Comparative proteomic study of phytotoxic effects of silver nanoparticles and silver ions on tobacco plants. Environ Sci Pollut Res 26, 22529–22550 (2019). https://doi.org/10.1007/s11356-019-05552-w
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DOI: https://doi.org/10.1007/s11356-019-05552-w