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AFLP and PCR-based markers linked to Rf3, a fertility restorer gene for S cytoplasmic male sterility in maize

Abstract

The Rf3 gene restores the pollen fertility disturbed by S male sterile cytoplasm. In order to develop molecular markers tightly linked to Rf3, we used amplified fragment length polymorphism (AFLP) technique with near isogenic lines (NILs) and bulk segregant analysis (BSA). A BC1F1 population from a pair of NILs with different Rf3 locus was constructed and 528 primer combinations was screened. A linkage map was constructed around the Rf3 locus, which was mapped on the distal region of chromosome 2 long arm with the help of SSR marker UMC2184. The closest marker E7P6 was 0.9 cM away from Rf3. Marker E3P1, 2.4 cM from Rf3, and E12M7, 1.8 cM from Rf3, were converted into a codominant CAPS and a dominant SCAR marker, and designated as CAPSE3P1 and SCARE12M7, respectively. These markers are useful for marker-assisted selection and map-based cloning of the Rf3 gene.

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Abbreviations

AFLP:

Amplified fragment length polymorphism

BF:

Fertile DNA bulks

BS:

Sterile DNA bulks

BSA:

Bulk segregant analysis

CAPS:

Cleaved amplified polymorphic sequence

CMS:

Cytoplasmic male sterility

E/M :

EcoRI/MseI

P/M :

PstI/MseI

P/E :

EcoRI/PstI

MAS:

Marker-assisted selection

NILs:

Near isogenic lines

ORF:

Open reading frames

PC:

Primer combinations

PPR:

Pentatricopeptide repeat

RAPD:

Random amplified polymorphic DNA

RFLP:

Restriction fragment length polymorphism

SCAR:

Sequence characterized amplified regions

SNP:

Singe nucleotide polymorphism

SSR:

Simple sequence repeat, microsatellite

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Acknowledgments

The authors thank Prof. Neil Forsberg for critical review of this manuscript. This research was supported by the National High Technique Foundation of China “863” (2004AA222160), “948” Fund in Ministry of Agronomy of China (201010), the State Key Basic Research and Development Plan of China “973” (2001CB1088).

Author information

Correspondence to Y. L. Zheng.

Additional information

Communicated by R. Hagemann

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Zhang, Z.F., Wang, Y. & Zheng, Y.L. AFLP and PCR-based markers linked to Rf3, a fertility restorer gene for S cytoplasmic male sterility in maize. Mol Genet Genomics 276, 162–169 (2006). https://doi.org/10.1007/s00438-006-0131-y

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Keywords

  • Maize
  • Fertility restorer
  • Near isogenic lines
  • Bulk segregant analysis
  • AFLP
  • PCR-based markers
  • Marker-assisted selection
  • Map-based cloning