Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)
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Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for F1-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called “functional” markers for allelic selection of the restorer-of-fertility gene (Rfo) in F1-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating F1-hybrid cultivar development in radish.
Abstract
Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for F1-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called “functional” markers for allelic selection of the restorer-of-fertility gene (Rfo) in F1-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating F1-hybrid cultivar development in radish.
참고문헌 (32)
- Albert B, "Evolution of the plant mitochondrial genome: dynamics of duplication and deletion of sequences", J. Mol. Evol., 46, pp.155~158 (1998)
- Andersen JR, "Functional markers in plants", Trends Plant Sci., 8, pp.554~560 (2003)
- Bentolila S, "A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants", Proc. Natl. Acad. Sci., 99, pp.10887~10892 (2002)
- Bonhomme S, "A 2.5 kb Nco I fragment of Ogura radish mitochondrial DNA is correlated with cytoplasmic male-sterility in Brassica cybrids", Curr. Genet., 19, pp.121~127 (1991)
- Brown GG, "The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats", Plant J., 35, pp.262~272 (2003)
- Budar F, "The nucleo- mitochondrial conflict in cytoplasmic male sterilities revised", Genetica, 117, pp.3~16 (2003)
- Cui X, "The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize", Science, 272, pp.1334~1336 (1996)
- Delourme R, "Characterization of the radish introgression carrying the Rfo restorer gene for the ogu-INRA cytoplasmic male sterility in rapeseed (Brassica napus L.)", Theor. Appl. Genet., 97, pp.129~134 (1998)
- Desloire S, "Identification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family", EMBO Rep., 4, pp.588~594 (2003)
- Fauron CMR, "Maize as a model of higher plant mitochondrial genome plasticity", Plant Sci., 112, pp.11~32 (1995)
- Gothandam KM, "OsPPR1, a pentatricopeptide repeat protein of rice is essential for the chloroplast biogenesis", Plant Mol. Biol., 58, pp.421~433 (2005)
- Grelon M, "Ogura cytoplasmic male-sterility (CMS)-associated orf138 is translated into a mitochondrial membrane polypeptide in male-sterile Brassica cybrids", Mol. Gen. Genet., 243, pp.540~547 (1994)
- Handa H, "The complete nucleotide sequence and RNA editing content of the mitochondrial genome of rapeseed (Brassica napus L.): comparative analysis of the mitochondrial genomes of rapeseed and Arabidopsis thaliana", Nucleic Acids Res., 31, pp.5907~5916 (2003)
- Hanson MR, "Interactions of mitochondrial and nuclear genes that affect male gametophyte development", Plant Cell, 16, pp.S154~S169 (2004)
- Kadowaki K, "A chimeric gene containing the 5’ portion of atp6 is associated with cytoplasmic male-sterility of rice", Mol. Genet. Genomics, 224, pp.10~16 (1990)
- Koizuka N, "Genetic characterization of a pentatricopeptide repeat protein gene, orf687, that restores fertility in the cytoplasmic male-sterile Kosena radish", Plant J., 34, pp.407~415 (2003)
- Koizuka N, "Genetic analysis of fertility restoration and accumulation of ORF125 mitochondrial protein in the kosena radish (Raphanus sativus cv. Kosena) and a Brassica napus restorer line", Theor. Appl. Genet., 100, pp.949~955 (2000)
- Komori T, "Map-based cloning of a fertility restorer gene, Rf-1, in rice (Oryza sativa L.)", Plant J., 37, pp.315~325 (2004)
- Kubo T, "The complete nucleotide sequence of the mitochondrial genome of sugar beet (Beta vulgaris L.) reveals a novel gene for tRNAcys (GCA)", Nucleic Acids Res., 28, pp.2571~2576 (2000)
- Lurin C, "Genome-wide analysis of Arabidopsis pentatricopeptide repeat proteins reveals their essential role in organelle biogenesis", Plant Cell, 16, pp.2089~2103 (2004)
- Makaroff CA, "Extensive mitochondrial specific transcription of the Brassica campestris mitochondrial genome", Nucleic Acids Res., 15, pp.5141~5156 (1987)
- Meyers BC, "Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis", Plant Cell, 15, pp.809~834 (2003)
- Nieuwhof M, "Cytoplasmic-genetic male sterility in radish (Raphanus sativus L.). Identification of maintainers, inheritance of male sterility and effect of environmental factors", Euphytica, 47, pp.171~177 (1990)
- Nivison HT, "Identification of a mitochondrial protein associated with cytoplasmic male sterility in petunia", Plant Cell, 1, pp.1121~1130 (1989)
- Notsu Y, "The complete sequence of the rice (Oryza sativa L.) mitochondrial genome: frequent DNA sequence acquisition and loss during the evolution of flowering plants", Mol. Genet. Genomics, 268, pp.434~445 (2002)
- Ogura H, "Studies on the new male sterility in Japanese radish, with special reference to the utilization of this sterility towards the practical raising of hybrid seeds", Mem Fac Agr Kagoshima Univ, 6, pp.39~78 (1968)
- Oldenburg DJ, "Mitochondrial DNA from the Liverwort Marchantia polymorpha: Circularly permuted linear molecules, head-to-tail concatemers, and a 5’ protein", J. Mol. Biol., 310, pp.549~562 (2001)
- Pelletier G, "Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion", Mol. Gen. Genet, 191, pp.244~250 (1983)
- Schnable PS, "The molecular basis of cytoplasmic male sterility and fertility restoration", Trends Plant Sci., 3, pp.175~180 (1998)
- Small ID, "The PPR motif- a TPR-related motif prevalent in plant organellar proteins", Trends Biochem. Sci., 25, pp.45~47 (2000)
- Unseld M, "The mitochondrial genome of Arabidopsis thaliana contains 57 genes in 366,924 nucleotides", Nature genet., 15, pp.57~61 (1997)
- Yamagishi H, "Molecular and biological studies on male-sterile cytoplasm in the Cruciferae. I. The origin and distribution of Ogura male-sterile cytoplasm in Japanese wild radishes (Raphanus sativus L.) revealed by PCR-aided assay of their mitochondrial DNAs", Theor. Appl. Genet., 87, pp.996~1000 (1994)