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Research Project: Towards Resilient Agricultural Systems to Enhance Water Availability, Quality, and Other Ecosystem Services under Changing Climate and Land Use

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Title: Adaption of microarray primers for iron transport and homeostasis gene expression in pseudomonas fluorescens exposed to nano iron

Author
item Fortuna, Ann Marie
item SINHA, SANJIVNI - North Dakota State University
item DAS, TONOY - North Dakota State University
item BEZBARUAH, ACHINTYA - North Dakota State University

Submitted to: MethodsX
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/5/2019
Publication Date: 4/30/2019
Citation: Fortuna, A., Sinha, S., Das, T.K., Bezbaruah, A.N. 2019. Adaption of microarray primers for iron transport and homeostasis gene expression in pseudomonas fluorescens exposed to nano iron. MethodsX. 6:1181-1187. https://doi.org/10.1016/j.mex.2019.04.006.
DOI: https://doi.org/10.1016/j.mex.2019.04.006

Interpretive Summary: Nanoparticles (<100 nm) are currently in use for biomedical, personal care, industrial, and environmental remediation applications. Therefore, assessing their impact on environmentally beneficial microorganisms such as Pseudomonas fluorescence is imperative. Nanomaterials such as to nanoscale zero-valent iron (NZVI) have different chemical properties and react differently than microscale (bulk) materials. Therefore, there is a need to develop and adapt current methods for use with nanomaterials in order to account for the unique properties that nanomaterials exhibit. The current methods were developed to address the needed modifications to existing protocols for use to evaluate NZVI. Specific methods may have to be developed for each nanomaterial tested.

Technical Abstract: Modifications were made to protocols used for PCR and culture based methods for the analysis of Pseudomonas fluorescens cells exposed to nanoscale zero-valent iron (NZVI), and atomic absorption spectrometric (AAS) analysis of iron in nutrient media. In furtherance of our understanding the effects of NZVI on this bacterium that chelates Fe by producing siderophores, we adapted sets of microarray primers used to quantify gene expression of pvdS and a bacterioferritin-associated ferredoxin gene for use in real-time quantitative reverse transcription (qRT-PCR) analysis. pvdS is one of a cluster of genes regulating the synthesis of the siderophore pyoverdine, also measured using chrome azrul S (CAS) plates. Pyoverdine bacterioferritin-associated ferredoxin is involved in mobilization of Fe from the protein (bacterioferritin B). Gene expression is dependent on the amount of Fe available to a bacterial cell. Therefore, reductions in the activity of these genes were measured after application of NZVI to P. fluorescens cultures.