Please use this identifier to cite or link to this item:
https://hdl.handle.net/11681/33388Full metadata record
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.author | Kennedy, Alan James, 1976- | - |
| dc.contributor.author | Smith, Natalie. | - |
| dc.contributor.author | Linan, Alexander. | - |
| dc.contributor.author | Kovacs. Laszlo. | - |
| dc.date.accessioned | 2019-07-11T19:04:26Z | - |
| dc.date.available | 2019-07-11T19:04:26Z | - |
| dc.date.issued | 2019-05 | - |
| dc.identifier.govdoc | ERDC/EL SR-19-3 | - |
| dc.identifier.uri | https://hdl.handle.net/11681/33388 | - |
| dc.identifier.uri | http://dx.doi.org/10.21079/11681/33388 | - |
| dc.description | Special Report | - |
| dc.description.abstract | Determining the toxicity of engineered nanomaterials (ENMs) to plants is important from an environmental perspective. Due to the absence of standardized phytotoxicity test methods for ENMs, a number of methods have been used and have produced inconsistent—and, consequently, unusable—results. This document describes a scientific operating procedure (SOP) to test the effects of plant chronic exposure to low doses of ENMs. The SOP is not specific to any particular ENM, but is designed to be applicable to any ENM that can be dispersed in water. Relying on Col-0, the highly homozygous wild-type laboratory strain of Arabidopsis, the method mitigates biological variance inherent in bioassay-based tests. In vitro growth conditions allow for the use of uniform synthetic growth medium and the exclusion of microbes. The detection of toxicity threshold is based on measuring the number of rosette leaves and the total exposed leaf area. If a physiological impact on the plant is detected, it is confirmed by the detection of ENM-induced changes in gene expression at the toxicity threshold to ensure that gene expression analysis is performed on plants grown at sublethal ENM concentrations. ENM-induced gene expression changes are then validated with the independent method of real-time quantitative PCR (qPCR). | en_US |
| dc.description.sponsorship | Environmental Consequences of Nanotechnologies (U.S.) | en_US |
| dc.description.tableofcontents | Abstract ................................................................................................................................................... ii Contents ................................................................................................................................................. iii Figures and Tables .................................................................................................................................. v Preface .................................................................................................................................................... vi Acronyms and Abbreviations ............................................................................................................... vii Unit Conversion Factors ...................................................................................................................... viii 1 Introduction ..................................................................................................................................... 1 1.1 Background .................................................................................................................... 1 1.2 Objective ........................................................................................................................ 3 1.3 Approach ........................................................................................................................ 3 2 Terminology ..................................................................................................................................... 5 2.2 Definitions ...................................................................................................................... 5 3 Materials and Apparatus ............................................................................................................... 6 3.1 Materials ........................................................................................................................ 6 3.2 Apparatus ....................................................................................................................... 6 4 Procedure ........................................................................................................................................ 8 4.1 Specimen preparation ................................................................................................... 8 4.1.1 Medium preparation ................................................................................................... 8 4.1.2 Seed sterilization ......................................................................................................... 9 4.1.3 Sowing, seed stratification, and plant culture ......................................................... 10 4.2 Toxicity gradient test .................................................................................................... 10 4.3 RNA-seq analysis ......................................................................................................... 11 4.3.1 RNA extraction .......................................................................................................... 11 4.3.2 RNA sample cleanup ................................................................................................ 12 4.3.3mRNA purification and complementary DNA (cDNA) synthesis ..................................... 13 4.3.4 cDNA sequencing ...................................................................................................... 13 4.4.2 qPCR .......................................................................................................................... 14 5 Reporting ....................................................................................................................................... 16 5.1 Analysis of results: ....................................................................................................... 16 5.1.1 Toxicity gradient test ................................................................................................. 16 5.1.2 RNA-seq analysis....................................................................................................... 16 5.2 Key results provided .................................................................................................... 18 5.2.1 Toxicity gradient test ................................................................................................. 18 5.2.2. RNA-seq Analysis ...................................................................................................... 19 5.2.3. qPCR validation ......................................................................................................... 19 5.3 Quality Assurance/Quality Control Considerations .................................................... 20 6 Conclusions ................................................................................................................................... 22 References ............................................................................................................................................ 23 Appendix: Notes and Supplementary Data ....................................................................................... 26 Report Documentation Page | - |
| dc.format.extent | 42 pages / 1.365 Mb | - |
| dc.format.medium | - | |
| dc.language.iso | en_US | en_US |
| dc.publisher | Environmental Laboratory (U.S.) | en_US |
| dc.publisher | Engineer Research and Development Center (U.S.) | - |
| dc.relation.ispartofseries | Special Report (Engineer Research and Development Center (U.S.)) ; no. ERDC/EL SR-19-3 | - |
| dc.rights | Approved for Public Release; Distribution is Unlimited | - |
| dc.source | This Digital Resource was created in Microsoft Word and Adobe Acrobat | - |
| dc.subject | Bioassay | en_US |
| dc.subject | Carbon nanomaterials | en_US |
| dc.subject | Engineered nanomaterial toxicity | en_US |
| dc.subject | ERDC NanoGRID | en_US |
| dc.subject | Filmetrics | en_US |
| dc.subject | Thin Film Analyzer | en_US |
| dc.subject | Toxicology | en_US |
| dc.subject | Spectral reflectance | en_US |
| dc.subject | Refractive index | en_US |
| dc.title | Bioassay to assess toxicity of water-dispersed engineered nanomaterials in plants; Scientific Operating Procedure Series : Toxicology (T) | en_US |
| dc.type | Report | en_US |
| Appears in Collections: | Special Report | |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| ERDC-EL SR-19-3.pdf | 1.4 MB | Adobe PDF |  View/Open |