Cell/Development/Systems

Abstract

CS-20-6 - Single-locus multi-hormone reporters for comprehensive plant phenotyping: a synthetic-biology approach

Monday, July 16
4:58 PM - 5:00 PM

Reductionist approaches have been traditionally used to understand how individual hormones carry out important functions in plant development, adaptation and survival. Nowadays, holistic approaches are arising as a powerful methodology to address the complexity of biological systems. In the field of hormone regulation, the contribution of different phytohormones to various processes should be addressed by monitoring the activity of multiple hormones simultaneously and with high temporal and spatial resolution. Even though a number of different methods have been developed for the detection and quantification of growth regulators in plants, none fulfill the requirements of both multiplexing and resolution. Herein, we are taking advantage of the power of the GoldenBraid (GB) gene assembly technology to enable combinatorial arrangement of multiple hormone transcriptional reporters to make a single-locus multi-hormone sensor: the hormometer. An ideal hormometer should consist of a single module comprised of multiple (up to nine) hormone-specific transcriptional reporters (HTR) for the nine major non-peptide plant hormones arranged in tandem. Each HTR unit contains five elements (phytobricks): a hormone-specific distal promoter, a core promoter (+5’UTR), a subcellular localization signal, a fluorescent protein coding sequence, and a terminator (+3’UTR). The GB modular DNA construction methods allow for the efficient generation of both individual transcriptional units (TUs) and multi-TU assembles in a matter of days. Our goal is to make the first hormometer (starting with one for just three hormones) and to characterize the resulting Arabidopsis and tomato stable transgenic lines. Towards this objective, we are generating the phytobrick collection of individual parts as well as assembling the single HTRs for all nine hormones. Functional validation of these HTRs is being performed in transient assays and, upon combining three HTRs and a resistance marker in a single module, will also be carried out in stably transformed plants.


 

Co-Authors

Jose Ascencio-Ibanñez – NCSU; Jose Alonso – NCSU; Anna Stepanova – NCSU

Josefina Patricia Fernandez. Moreno

PhD
NCSU

Presentation(s):

Send Email for Josefina Patricia Moreno


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CS-20-6 - Single-locus multi-hormone reporters for comprehensive plant phenotyping: a synthetic-biology approach



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