Quantification of transgene expression in maize (Zea mays L.) throughout the vegetation period

Publication Type:Journal Article
Year of Publication:2011
Authors:Bakó, A, Gell, G, Balázs, E
Journal:Plant Breeding
Volume:130
Issue:1
Date Published:2011
ISBN Number:1439-0523
Keywords:gene expression, maize, quantitative PCR, RNA extraction, transgene
Abstract:

With 3 figures and 1 tableAbstract Evaluation of transgenic plants during their development is of major importance for breeders. To facilitate this evaluation, we applied a real-time, reverse transcription polymerase chain reaction method to maize tissues that allows quantitative expression profiling of transgenes. To obtain consistent results, sampling, transport, storage and extraction protocols were standardized for a wide range of samples (leaves, stems, roots, pollen and kernels) and phenological stages (from the 4–6 leaves stage to physiological maturity). RNA integrity was monitored by Agilent 2100 Bioanalyzer to ensure that decomposed RNA samples were excluded from further study. A simple and effective SYBR Green methodology was applied for relative quantification of transgene expression. Several endogenous controls were tested to achieve the best possible results. Quantitave results presented illustrate the application of our protocol to the quantification of cry3Bb1 (encoding corn root worm resistance) expression in MON88017 event derived transgenic maize lines. The methodology described here can be routinely performed for the quantification of gene expression in maize.

URL:http://dx.doi.org/10.1111/j.1439-0523.2010.01804.x
Short Title:Plant Breeding
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Scratchpads developed and conceived by (alphabetical): Ed Baker, Katherine Bouton Alice Heaton Dimitris Koureas, Laurence Livermore, Dave Roberts, Simon Rycroft, Ben Scott, Vince Smith