|Kupper, Hendrik - CORNELL UNVIERSITY|
|Seib, Laura - CORNELL UNIVERSITY|
Submitted to: American Society of Plant Biologists Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: March 19, 2004
Publication Date: July 24, 2004
Citation: Kupper, H., Seib, L.O., Kochian, L.V. 2004. Analysis of zinc metabolism in thlaspi and arabidopsis by observing zn transporter gene expression in situ and in vivo in comparison to photosynthesis measured with fluorescence kinetic microscopy. American Society of Plant Biologists Annual Meeting. p. 190. Technical Abstract: Heavy metal uptake by plants is an important process both in terms of plant stress physiology and human nutrition. Little is known, however, about the regulation and cellular expression patterns of heavy metal transporters in plants. In this work we use a novel in situ hybridisation technique as well as YFP and DsRed promoter-reporter constructs to elucidate expression patterns (mRNA levels) of several members of the ZNT/ZIP gene family in Thlaspi caerulescens, Thlaspi arvense, Arabidopsis thaliana and Arabidopsis halleri. In addition to observing tissue- and age-dependent differences in gene expression patterns, the quantitative nature of the applied in situ hybridisation technique furthermore allowed for an analysis of the response to different metal treatments. In addition, our use of whole mounts combined with optical sectioning (instead of microtome sectioning) dramatically increased the throughput of our protocol compared to conventional in situ hybridisation methods. YFP and DsRed promoter-reporter gene constructs were used to follow the above-mentioned aspects of transporter gene regulation also in vivo. For several reasons this approach is in principle less quantitative that our new in situ hybridisation technique. But in contrast to in situ hybridisation, it allows for a direct comparison of ZNT/ZIP transcription with photosynthesis in the same cells, because both fluorescent proteins and photosynthesis can be measured concurrently in vivo using the recently developed technique of Fluorescence Kinetic Microscopy.