| Title: | High-throughput screening for in planta characterization of VOC biosynthetic genes by PTR-ToF-MS |
| Author(s): | Li M; Cappellin L; Xu J; Biasioli F; Varotto C; |
| Address: | "Department of Biodiversity and Molecular Ecology, Research and Innovation Centre, Fondazione Edmund Mach, 38010, San Michele all' Adige, TN, Italy. Dipartimento di Scienze Chimiche, Universita degli Studi di Padova, Via Marzolo 1, 35121, Padua, Italy. Dipartimento di Biologia, Universita di Padova, Viale G. Colombo 3, 35121, Padua, Italy. Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach, 38010, San Michele all' Adige, TN, Italy. Department of Biodiversity and Molecular Ecology, Research and Innovation Centre, Fondazione Edmund Mach, 38010, San Michele all' Adige, TN, Italy. claudio.varotto@fmach.it" |
| DOI: | 10.1007/s10265-019-01149-z |
| ISSN/ISBN: | 1618-0860 (Electronic) 0918-9440 (Print) 0918-9440 (Linking) |
| Abstract: | "Functional characterization of plant volatile organic compound (VOC) biosynthetic genes and elucidation of the biological function of their products often involve the screening of large numbers of plants from either independent transformation events or mapping populations. The low time resolution of standard gas chromatographic methods, however, represents a major bottleneck for in planta genetic characterization of VOC biosynthetic genes. Here we present a fast and highly-sensitive method for the high-throughput characterization of VOC emission levels/patterns by coupling a Proton Transfer Reaction Time-of-Flight Mass Spectrometer to an autosampler for automation of sample measurement. With this system more than 700 samples per day can be screened, detecting for each sample hundreds of spectrometric peaks in the m/z 15-300 range. As a case study, we report the characterization of VOC emissions from 116 independent Arabidopsis thaliana lines transformed with a putative isoprene synthase gene, confirming its function also when fused to a C-terminal 3xFLAG tag. We demonstrate that the method is more reliable than conventional characterization of transgene expression for the identification of the most highly isoprene-emitting lines. The throughput of this VOC screening method exceeds that of existing alternatives, potentially allowing its application to reverse and forward genetic screenings of genes contributing to VOC emission, constituting a powerful tool for the functional characterization of VOC biosynthetic genes and elucidation of the biological functions of their products directly in planta" |
| Keywords: | Gas Chromatography-Mass Spectrometry Mass Spectrometry Protons Volatile Organic Compounds/*metabolism Arabidopsis thaliana Genetic screening Isoprene Ptr-ms Time-of-flight VOCs; |
| Notes: | "MedlineLi, Mingai Cappellin, Luca Xu, Jia Biasioli, Franco Varotto, Claudio eng 659315/H2020 Marie Sklodowska-Curie Actions/ Japan 2019/11/09 J Plant Res. 2020 Jan; 133(1):123-131. doi: 10.1007/s10265-019-01149-z. Epub 2019 Nov 7" |
