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KMID : 0578320060210030411
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Molecules and Cells
2006 Volume.21 No. 3 p.411 ~ p.417
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Determination of Cytoplasmic Male Sterile Factors in Onion Plants (Allium cepa L.) Using PCR-RFLP and SNP Markers
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Cho Kwang-Soo
Yang Tae-Jin
Hong Su-Young
Kwon Young-Seok
Woo Jong-Gyu
Park Hyo-Guen
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Abstract
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We have developed a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) marker that can distinguish male-fertile (N) and male-sterile (S) cytoplasm in onions. The PCR-RFLP marker was located in a chloroplast psbA gene amplicon. Digesting the amplicons from different cytoplasm-containing varieties with the restriction enzyme MspI revealed that N-cytoplasm plants have a functional MspI site (CCGG), whereas the S-cytoplasm plants has a substitution in that site (CTGG), and thus no MspI target. The results obtained using this PCR-RFLP marker to distinguish between cytoplasmic male sterile factors in 35 onion varieties corresponded with those using a CMS-specific sequence-characterized amplified region (SCAR) marker. Moreover, the PCR-RFLP marker can identify N- ot S-cytoplasms in DNA sample mixtures in which they are in up to a 10-fold minority, indicating that use of the marker has high diagnostic precision. We also demonstrated the usefulness of the SNP detected in the psbA gene for high-throughput discrimination of CMS factors using Real-time PCR and a TaqMan probe assay.
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KEYWORD
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Cytoplasmic Male Sterility (CMS), Markerassisted Breeding (MAB), PCR-RFLP, Real-Time PCR, Sequence Characterized Amplified Region (SCAR) Marker
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