Marker Assisted Selection in Wheat

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Disease resistance. Fusarium Head Blight (Scab)

Contributed by Jim Anderson

Microsatellite markers Xgwm493 and Xgwm533

DNA Extraction: chloroform-isoamyl alcohol (24:1) extraction procedure (detailed protocol).

Primers for Xgwm493-3B

Left primer  5'- TTC CCA TAA CTA AAA CCG CG -3'
Right primer 5'- GGA ACA TCA TTT CTG GAC TTT G -3'’

Primers for Xgwm533-3B

Left primer  5'- AAG GCG AAT CAA ACG GAA TA -3'
Right primer 5'- GTT GCT TTA GGG GAA AAG CC -3'

This primers were developed by Röder et. al. (Genetics 149: 2007-2023, August 1998).

PCR amplification mix:

• Template DNA (15ng/µL)	3.0 µL
• 10X PCR Buffer II	1.0 µL
• MgCl₂ (25 mM)	0.6 µL
• dNTPs (1.25 mM)	1.6 µL
• Forward Primer (1 µM)	1.0 µL
• Reverse Primer (1 µM)	1.0 µL
• Amplitaq Gold (5U/µL)	0.05 µL
• HPLC H₂O	1.75 µL
• Total	10.0 µL

PCR Program:

94°C for 10 minutes
Then 35 cycles of 94°C for 1 minute
60°C for 1 minute
72°C for 2 minutes
Final extension of 72°C for 10 minutes
Then 4°C hold.

Electrophoresis and expected products: Acrylamide gel (6.5%) with formamide (detailed protocol). Expected PCR product size: 290 and 140 bp for gwm493 and gwm533 primers, respectively.

Other markers: The BARC project has developed and mapped a set of microsatellite markers. Some of them are located located close to the peak of the QTL for FHB resistance from Sumai 3. This markers are: XBARC75, XBARC87 and XBARC73. You can find the primer sequences and the restrictions that apply for their use following this link.

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