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

• Published in: Journal of the science of food and agriculture Vol. 88; no. 13; pp. 2253 - 2264
• Main Authors: Cullen, Danny W, Squire, Geoffrey R, McNicol, Jim W, Jacobs, Jennifer H, Osborne, Juliet L, Ford, Linda, Ramsay, Gavin, Scrimgeour, Charlie, Young, Mark W
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-10-2008; Wiley; John Wiley and Sons, Limited
• Subjects: Biological and medical sciences; Brassica napus; Chromatography; Comparative analysis; cross pollination; erucic acid; fatty acid composition; fields; Food industries; Food science; Fundamental and applied biological sciences. Psychology; gas chromatography; gene flow; high erucic acid oilseed rape; low erucic acid oilseed rape; oilseed rape; Plant reproduction; pollen flow; polymerase chain reaction; Rape plants; rapeseed; rapeseed oil; real-time polymerase chain reaction; real-time qPCR; spatial variation; Validity; varieties; Scotland; Oilseed rape; Validation; gene flow; Monounsaturated fatty acid; Lipids; Erucic acid; Real time; real-time qPCR; Polymerase chain reaction; cross-pollination; Gas chromatography; Gene; Development; Molecular biology; Variety; Quantitative analysis
• ISSN: 0022-5142; 1097-0010
• DOI: 10.1002/jsfa.3340

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 1 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. Copyright