Multiplexed PCR to screen for a major QTL carrying fusarium head blight resistance in Sumai-3 wheat

Abstract

After the initial identification of microsatellites linked to economically important traits of interest, the additional investment to simplify the screening procedure for more routine use is of interest to plant breeders. The objective was to simplify an acrylamide gel based marker-assisted selection (MAS) method to facilitate high-through put screening for Qfhs.ndsu-3BS, a major quantitative trait locus carrying fusarium head blight (FHB) resistance in Sumai-3 wheat (Triticum aestivum L.). Method simplifications included incorporating the use of (1) 96-well DNA extractions, (2) multiplexed PCR reactions using microsatellite primers gwm493 and gwm533, and (3) agarose gels. Our modified FHB-MAS method was verified by screening six resistant (Sumai-3, ND2710, McVey, BacUp, HY644, and Alsen) and 52 susceptible parents and subsequently used to screen 5567 common wheat breeding lines developed from Sumai-3 derivatives. This simple and rapid method allows for the screening of 1000 lines per week, which can be used to skew segregating populations towards more resistant types and thereby advance primarily early and advanced generation lines that carry at least the major FHB QTL.