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A PCR Marker for Growth Habit in Common Wheat Based on Allelic Variation at the VRN-A1 Gene

^a Dep. of Plant Science, Montana State Univ., Bozeman, MT 59718
^b Dep. of Agronomy and Range Science, Univ. of California, Davis, CA 95616

View larger version (25K): [in a new window]   Fig. 1. PCR amplification of primers VA1-F and VA1-R. DNAs were digested with restriction enzyme HinfI. Normal Chinese Spring in the first lane is followed by nulli-tetrasomic lines missing chromosomes 5A, 5B, and 5D, respectively. Note that there is no amplification product in the nulli-5A line, confirming that the primers are A-genome specific. The band size after restriction with HinfI was 690 bp as predicted by the A genome sequence.

View larger version (40K): [in a new window]   Fig. 2. Consensus sequences for winter and spring genotypes of the region amplified by VRN-A1-specific primers (indicated in bold-italic letters at the beginning and end of the sequence). Dots in the sequence from the spring varieties indicate identical bases with the sequences from the winter varieties. The three differences found between the ‘s’ and ‘w’ haplotypes are indicated by bold letters. AciI restriction sites are underlined. The location of the allele specific primer 361W-F is indicated in underlined italic letters.

View larger version (18K): [in a new window]   Fig. 3. PCR product amplified by VRN-A1-specific primers and digested with restriction enzyme AciI. The growth habit of each variety is indicated below the photograph. The PCR product from the winter varieties is 76 bp larger than the PCR product from the spring varieties, which have an additional AciI restriction site. The varieties used in this figure except HiLine are not part of Table 1: H-HiLine, E-Era, V-Vanguard, F-Federation, Re-Red Egyptian, R-Rocky, Mt-MTW94410, R-Reeder, Ru-Russ, No-Norstar, P-Pilot, Ph-Pronghorn, 3-PI372129.

View larger version (26K): [in a new window]   Fig. 4. PCR products from F2 plants from the cross Newana x Redwin amplified by IRDye800-labeled primers 361SF and VA1-R and run on a Licor DNA Analyzer. Products were digested with restriction enzyme AciI to confirm that they originated in the A genome. Note that the specific primers for the ‘s’ allele amplified a product only from the 20 spring plants (1–20). The band is absent in the 10 winter individuals (21–30). Digestion with AciI produced the expected band of 116 bp, confirming the presence of a different ‘s’-specific polymorphism in the PCR products. M indicates the molecular weight marker.