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PCR-based markers for the powdery mildew resistance gene Pm4a in wheat

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Abstract

Gene tagging is the basis of marker-assisted selection and map-based cloning. To develop PCR-based markers for Pm4a, a dominant powdery mildew resistance gene of wheat, we surveyed 46 group 2 microsatellite markers between Pm4a near-isogenic line (NIL) CI 14124 and the recurrent parent Chancellor (Cc). One of the markers, gwm356, detected polymorphism and was used for genotyping an F2 population of 85 plants derived from CI 14124 × Cc. Linkage mapping indicated that Xgwm356 was linked to Pm4a at a distance of 4.8 cM. To identify more PCR-based markers for Pm4a, we also converted the restriction fragment length polymorphism marker BCD1231 linked to it into a sequence-tagged site (STS) marker. The STS primer designed based on the end sequences of BCD1231 amplified an approximately 1.6-kb monomorphic band in both parents. Following digestion of the products with the four-cutter enzymes HaeIII and MspI, it was discovered that the band from CI 14124 consisted of at least two products, one of which had a digestion pattern different from the band from Cc. In the F2 population, the cleaved polymorphism revealed by the STS marker between the parents co-segregated with powdery mildew resistance. To design Pm4a-specific PCR markers, the 1.6-kb band from Cc and the fragment associated with Pm4a in CI 14124 were sequenced and compared. Based on these sequences a new PCR marker was identified, which detected a 470-bp product only in the Pm4a-containing lines. These PCR-based markers provide a cost-saving option for marker-assisted selection of Pm4a.

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Acknowledgements

This project was partially supported by the Outstanding Youth Fund (no. 30025030) of NSFC, the ‘863’ project 2003AA207100, and ‘2000’ Trans-century Talent Development Program of MOE, China. We thank X.W. Gu and L. Xiong for their help in resistance screening.

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Authors and Affiliations

  1. National Key Lab of Crop Genetics and Germplasm Enhancement, College of Agricultural Sciences, Nanjing Agricultural University, 210095, Jiangsu, China

    Z.-Q. Ma & J.-B. Wei

  2. Lixiahe Agricultural Research Institute, 225002, Yangzhou, China

    S.-H. Cheng

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  1. Z.-Q. Ma
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  2. J.-B. Wei
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  3. S.-H. Cheng
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Correspondence to Z.-Q. Ma.

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Communicated by F. Salamini

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Ma, ZQ., Wei, JB. & Cheng, SH. PCR-based markers for the powdery mildew resistance gene Pm4a in wheat. Theor Appl Genet 109, 140–145 (2004). https://doi.org/10.1007/s00122-004-1605-0

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  • DOI: https://doi.org/10.1007/s00122-004-1605-0

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