Development and validation of gas chromatography and real-time quantitative PCR for the quantification of landscape-scale gene flow from varieties of high erucic acid (HEAR) oilseed rape

A - Papers appearing in refereed journals

Cullen, D. W., Squire, G. R., Mcnicol, J. W., Jacobs, J. H., Osborne, J. L., Ford, L., Ramsay, G., Scrimgeour, C. and Young, M. W. 2008. Development and validation of gas chromatography and real-time quantitative PCR for the quantification of landscape-scale gene flow from varieties of high erucic acid (HEAR) oilseed rape. Journal of the Science of Food and Agriculture - jsfa. 88 (13), pp. 2253-2264. https://doi.org/10.1002/jsfa.3340

AuthorsCullen, D. W., Squire, G. R., Mcnicol, J. W., Jacobs, J. H., Osborne, J. L., Ford, L., Ramsay, G., Scrimgeour, C. and Young, M. W.
Abstract

BACKGROUND: High erucic acid oilseed rape (HEAR) was tested as a source crop for estimates of regional geneflow. Two methods to detect HEAR in low erucic acid oilseed rape (LEAR) were compared: real-time quantitative PCR (qPCR) and gas chromatography (GC). RESULTS: Fields (2.5 ha) of a LEAR variety (0.028% EA) in Tayside and Hertfordshire were juxtaposed adjacent to and I km distant from a HEAR (44% EA) field. The LEAR variety was a varietal association to ensure high cross-pollination (CP). The methods were highly correlated, measuring between 30% and 0.5% CP. However, the qPCR method became unreliable below 0.5% CP, whereas GC was robust enough to detect raised EA equivalent to one F1 seed in 500 (0.2%). A statistical mixture model was fitted to the distributions of EA in samples in order to assign a CP value to each 500-seed sample. Declines of CP from 30% to < 1%, and EA from 5% to 0.2%, with distance up to 150 m in the near fields was best fitted with a power function. The combined mean EA for both far fields was 0.11%, well above the background LEAR value of 0.028%, and mean CP was 0.36%. CONCLUSIONS: The GC method of detection raised %EA should be a reliable and high-throughput means of estimating %CP between fields, provided the %EA of single F1 seed in receptor fields is measured to confirm the presence of F1 seed. (c) 2008 Society of Chemical Industry.

KeywordsAgriculture, Multidisciplinary; Chemistry, Applied; Food Science & Technology
Year of Publication2008
JournalJournal of the Science of Food and Agriculture - jsfa
Journal citation88 (13), pp. 2253-2264
Digital Object Identifier (DOI)https://doi.org/10.1002/jsfa.3340
Open accessPublished as non-open access
FunderCentral Science Laboratory - CSL MAFF
ADAS
NIAB
NERC - Natural Environment Research Council - Centre for Ecology Hydrology - CEH
Funder project or codeSEF
Project: 4554
Functional biodiversity: mechanisms by which plant and invertebrate communities function in the arable ecosystem
ISSN00225142
0022-5142
PublisherWiley

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