Fast tracking MAS through a new tissue sampling technique – An investigation

Abstract

Recent developments in molecular marker techniques have allowed the throughput of marker assisted selection (MAS) in breeding programs to increase rapidly over the past few years. This is largely a result of more efficient DNA extraction methods and the fluorescent-based detection of multiplexed PCR markers on semi-automated DNA fragment analysers. To streamline and facilitate the screening of larger populations a more efficient tissue sampling technique and planting strategy is required. Glasshouse space and seed / data management has now become one of the limiting steps in marker screening. This paper investigates the opportunity to implement a new tissue sampling technique that would alleviate limiting factors such as glasshouse space. Barley seeds are planted, grown and harvested in the laboratory in a 96 well format. The DNA extraction is performed on root tissue following freezing to capture a portion of the root tissue. By harvesting only some of the roots, the seedling is left viable. Hence after MAS, lines carrying the desired alleles can be transplanted in the glasshouse to reach maturity. This technique has been successfully used in Arabidopsis and rice (5).