Abstract
The Polima (pol) system of cytoplasmic male sterility (CMS) and its fertility restorer gene Rfp are used in hybrid rapeseed production in Brassica napus. To facilitate map-based cloning of the Rfp gene, we have successfully transferred the pol cytoplasm and Rfp from the amphidiploid B. napus to the diploid species B. rapa and generated a doubled haploid pol cytoplasm B. rapa population that segregates for the Rfp gene. This was achieved through interspecific crosses, in vitro rescue of hybrid embryos, backcrosses, and microspore culture. Male fertility conditioned by Rfp was shown to co-segregate in this population with Rfp-specific mitochondrial transcript modifications and with DNA markers previously shown to be linked to Rfp in B. napus. The selfed-progeny of one doubled haploid plant were confirmed to be characteristic B. rapa diploids by cytogenetic analysis. Clones recovered from a genomic library derived from this plant line using the RFLP probe cRF1 fell into several distinct physical contigs, one of which contained Rfp-linked polymorphic restriction fragments detected by this probe. This indicates that chromosomal DNA segments anchored in the Rfp region can be recovered from this library and that the library may therefore prove to be a useful resource for the eventual isolation of the Rfp gene.
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Abbreviations
- CMS:
-
cytoplasmic male sterility
- DH:
-
doubled haploid
- h:
-
hour
- min:
-
minute
- kb:
-
kilobase
- PPR:
-
pentatricopeptide
- Rf:
-
restorer of fertility
References
G.G. Brown N. Formanová H. Jin R. Wargachuk C. Dendy P. Patil M. Laforest J. Zhang W.Y. Cheung B.S. Landry (2003) ArticleTitleThe radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats Plant J. 35 262–272 Occurrence Handle12848830 Occurrence Handle1:CAS:528:DC%2BD3sXmvFSmu7Y%3D Occurrence Handle10.1046/j.1365-313X.2003.01799.x
G.G. Brown (1999) ArticleTitleUnique aspects of cytoplasmic male sterility and fertility restoration in Brassica napus J. Heredity 90 351–356 Occurrence Handle1:CAS:528:DyaK1MXjvVKmsLc%3D Occurrence Handle10.1093/jhered/90.3.351
F. Budar P. Touzet R. Paepe ParticleDe (2003) ArticleTitleThe nucleo-mitochondrial conflict in cytoplasmic male sterilities revisited Genetica 117 3–16 Occurrence Handle12656568 Occurrence Handle1:CAS:528:DC%2BD3sXhtFGjs70%3D Occurrence Handle10.1023/A:1022381016145
A.C. Cavell D.J. Lydiate I.A. Parkin C. Dean M. Trick (1998) ArticleTitleCollinearity between a 30-centimorgan segment of Arabidopsis thaliana chromosome 4 and duplicated regions within the Brassica napus genome Genome 41 62–69 Occurrence Handle9549059 Occurrence Handle1:CAS:528:DyaK1cXisVChu7o%3D Occurrence Handle10.1139/gen-41-1-62
P.V. Chuong K.P. Pauls W.D. Beversddorf (1988) ArticleTitleHigh efficiency embryogenesis in male sterile plants of Brassica napus through microspore culture Can. J. Bot. 66 1676–1680
S.J. Dellaporta J. Wood J.B. Hicks (1983) ArticleTitleA plant mini-DNA preparation: version II Plant Mol. Biol. Rep. 1 19–21 Occurrence Handle1:CAS:528:DyaL2cXksFWhtrk%3D
S. Desloire H. Gherbi W. Laloui S. Marhadour V. Clouet L. Cattolico C. Falentin S. Giancola M. Renard F. Budar I. Small M. Caboche R. Delourme A. Bendahmane (2003) ArticleTitleIdentification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family EMBO Rep. 4 588–594 Occurrence Handle12740605 Occurrence Handle1:CAS:528:DC%2BD3sXktFOgtrs%3D Occurrence Handle10.1038/sj.embor.embor848
Z. Fan B.R. Stefansson (1986) ArticleTitleInfluence of temperature on sterility of two cytoplasmic male-sterility systems in rape (Brassica napus L.) Can. J. Plant Sci. 66 221–227 Occurrence Handle10.4141/cjps86-035
G.H. Fang P.B.E. McVetty (1989) ArticleTitleInheritance of male fertility restoration and allelism of restorer genes for the Polima cytoplasmic male sterility system in oilseed rape Genome 32 1044–1047
A.M.R. Ferrie W.A. Keller (1995) Microspore culture for haploid plant production O.L. Gamborg G.C. Phillips (Eds) Fundamental Methods of Plant Cell, Tissue and Organ Culture and Laboratory Operations Springer Verlag Berlin 155–164
V. Grant (1975) Genetics of Flowering Plants Columbia University Press New York
H. Handa K. Nakajima (1992) ArticleTitleDifferent organization and altered transcription of the mitochondrial atp6 gene in the male-sterile cytoplasm of rapeseed (Brassica napus L.) Curr. Genet. 21 153–159 Occurrence Handle1533177 Occurrence Handle1:CAS:528:DyaK3sXhvVCltrs%3D Occurrence Handle10.1007/BF00318475
M. Hanson S. Bentolila (2004) ArticleTitleInteractions of mitochondrial and nuclear genes that affect male gametophyte development Plant Cell 16 S164–S169 Occurrence Handle10.1105/tpc.015966
M. Jean G.G. Brown B.S. Landry (1997) ArticleTitleGenetic mapping of nuclear fertility restorer genes for the ‘Polima’ cytoplasmic male sterility in canola (Brassica napus L.) using DNA markers Theor. Appl. Genet. 95 321–328 Occurrence Handle1:CAS:528:DyaK2sXmtFGrtrY%3D Occurrence Handle10.1007/s001220050566
N. Koizuka R. Imai H. Fujimoto T. Hayakawa Y. Kimura J. Kohno-Murase T. Sakai S. Kawasaki J. Imamura (2003) ArticleTitleGenetic characterization of a pentatricopeptide repeat protein gene, orf687: that restores fertility in the cytoplasmic male-sterile Kosena radish Plant J. 34 407–415 Occurrence Handle12753581 Occurrence Handle1:CAS:528:DC%2BD3sXkslOjtb4%3D Occurrence Handle10.1046/j.1365-313X.2003.01735.x
U. Lagercrantz (1998) ArticleTitleComparative mapping between Arabidopsis thaliana and Brassica nigra indicates that Brassica genomes have evolved through extensive genome replication accompanied by chromosome fusions and frequent rearrangements Genetics 150 1217–1228 Occurrence Handle9799273 Occurrence Handle1:CAS:528:DyaK1cXnsFCntb0%3D
T.H. Lan T.A. DelMonte K.P. Reischmann J. Hyman S.P. Kowalski J. McFerson S. Kresovich A.H. Paterson (2000) ArticleTitleAn EST-enriched comparative map of Brassica oleracea and Arabidopsis thaliana Genome Res. 10 776–788 Occurrence Handle10854410 Occurrence Handle1:CAS:528:DC%2BD3cXkt1eisLs%3D Occurrence Handle10.1101/gr.10.6.776
E.S. Lander P. Green J. Abrahamson A. Barlow M.J. Daly S.E. Lincoln L. Newburg (1987) ArticleTitleMAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations Genomics 1 174–181 Occurrence Handle3692487 Occurrence Handle1:CAS:528:DyaL1cXhsVCksrk%3D Occurrence Handle10.1016/0888-7543(87)90010-3
B.S. Landry N. Hubert T. Etoh J.J. Harada S.E. Lincoln (1991) ArticleTitleA genetic map of Brassica napus based on restriction fragment length polymorphisms detected with expressed DNA sequences Genome 34 543–552 Occurrence Handle1:CAS:528:DyaK38XhsVGitbo%3D
Y. L’Homme R.J. Stahl X.-Q. Li A. Hameed G.G. Brown (1997) ArticleTitleBrassica nap cytoplasmic male sterility is associated with expression of a mtDNA region containing a chimeric gene similar to the pol CMS-associated orf224 gene Curr. Genet. 31 325–335 Occurrence Handle9108140 Occurrence Handle1:CAS:528:DyaK2sXjtlKmurc%3D Occurrence Handle10.1007/s002940050212
Y. L’Homme G.G. Brown (1993) ArticleTitleOrganizational differences between cytoplasmic male sterile and male fertile Brassica mitochondrial genomes are confined to a single transposed locus Nucl. Acids Res. 21 1903–1909 Occurrence Handle8388101 Occurrence Handle1:CAS:528:DyaK3sXktVagsr8%3D
X.Q. Li N. Formanova D.B. Zhu G.G. Brown (1996) ArticleTitleAgrobacterium-mediated transformation of oilseed rape (Brassica napus) with cosmid-type binary vectors Progress in Natural Sci. 6 568–576 Occurrence Handle1:CAS:528:DyaK28XmsVyjtLc%3D
X.Q. Li M. Jean B.S. Landry G.G. Brown (1998) ArticleTitleRestorer genes for different forms of Brassica cytoplasmic male sterility map to a single nuclear locus that modifies transcripts of several mitochondrial genes Proc. Natl. Acad. Sci. USA 95 10032–10037 Occurrence Handle9707595 Occurrence Handle1:CAS:528:DyaK1cXlsFWjur8%3D Occurrence Handle10.1073/pnas.95.17.10032
R. Menassa Y. L’Homme G.G. Brown (1999) ArticleTitlePost-transcriptional and developmental regulation of a CMS-associated mitochondrial gene region by a nuclear restorer gene Plant J. 17 491–499 Occurrence Handle10205904 Occurrence Handle1:CAS:528:DyaK1MXisVKmsbo%3D Occurrence Handle10.1046/j.1365-313X.1999.00397.x
T. Murashige F. Skoog (1962) ArticleTitleA revised medium for rapid growth and bioassay with tobacco tissue cultures Physiol. Plant 15 473–497 Occurrence Handle1:CAS:528:DyaF3sXksFKm Occurrence Handle10.1111/j.1399-3054.1962.tb08052.x
N.E. Olszewski F.B. Martin F.M. Ausubel (1988) ArticleTitleSpecialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene in Nicotiana tabacum Nucl. Acids Res. 16 10765–10782 Occurrence Handle3060849 Occurrence Handle1:CAS:528:DyaL1MXntVChtQ%3D%3D
C. O’Neill I. Bancroft (2000) ArticleTitleComparative physical mapping of segments of the genome of Brassica oleracea var alboglabra that are homeologous to sequenced regions of the chromosomes 4 and 5 of Arabidopsis thaliana Plant J. 23 233–243 Occurrence Handle10929117 Occurrence Handle1:CAS:528:DC%2BD3cXmtFOnsLw%3D Occurrence Handle10.1046/j.1365-313x.2000.00781.x
D. Rana T. Boogaart Particlevan den C.M. O’Neil L. Hynes E. Bent L. Macpherson J.Y. Park Y.P. Lim I. Bancroft (2004) ArticleTitleConservation of the microstructure of genome segments in Brassica napus and its diploid derivatives Plant J. 40 725–733 Occurrence Handle15546355 Occurrence Handle1:CAS:528:DC%2BD2MXjvFWhuw%3D%3D Occurrence Handle10.1111/j.1365-313X.2004.02244.x
M. Singh G.G. Brown (1991) ArticleTitleSuppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitochondrial gene region Plant Cell 3 1349–1362 Occurrence Handle1840901 Occurrence Handle1:CAS:528:DyaK3sXhtFyjsLs%3D Occurrence Handle10.1105/tpc.3.12.1349
M. Singh N. Hamel R. Menassa X.Q. Li B. Young M. Jean B.S. Landry G.G. Brown (1996) ArticleTitleNuclear genes associated with a single Brassica CMS restorer locus influence transcripts of three different mitochondrial gene regions Genetics 143 505–516 Occurrence Handle8722799 Occurrence Handle1:CAS:528:DyaK28XjtVKjs7Y%3D
I.D. Small N. Peeters (2000) ArticleTitleThe PPR motif - a TPR-related motif prevalent in plant organellar proteins Trends Biochem Sci. 25 46–47 Occurrence Handle10664580 Occurrence Handle1:CAS:528:DC%2BD3cXht1Krsr4%3D Occurrence Handle10.1016/S0968-0004(99)01520-0
J.K. Verma Y.S. Sodhi A. Mukhopadhyay N. Arumugam V. Gupta D. Pental A.K. Pradhan (2000) ArticleTitleIdentification of stable maintainer and fertility restorer lines for ‘Polima’ CMS in Brassica campestris Plant Breeding 119 90–92 Occurrence Handle10.1046/j.1439-0523.2000.00430.x
H.M. Wang T. Ketela W.A. Keller S.C. Gleddie G.G. Brown (1995) ArticleTitleGenetic correlation of the orf224/atp6 gene region with Polima CMS in Brassica somatic hybrids Plant Mol. Biol. 27 801–807 Occurrence Handle7727756 Occurrence Handle1:CAS:528:DyaK2MXlt1Wnu7c%3D Occurrence Handle10.1007/BF00020232
J.H. Weiss (1999) Plating and transferring plasmid and cosmid libraries F.M. Ausubel R. Brent R.E. Kingston D.D. Moore J.G. Seidman J.A. Smith K. Struhl (Eds) Short Protocols in Molecular Biology John Wiley and Sons New York 64–65
P.H. Williams C.B. Hill (1986) ArticleTitleRapid-cycling populations of Brassica Science 232 1385–1389 Occurrence Handle17828914
U. Witt S. Hansen M. Albaum W.O. Abel (1991) ArticleTitleMolecular analyses of the CMS-inducing Polima cytoplasm of Brassica napus L Curr. Genet. 19 323–327 Occurrence Handle1:CAS:528:DyaK3MXks1GrtLY%3D Occurrence Handle10.1007/BF00355063
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Nataša Formanová and Xiu-Qing Li: These two authors contributed equally to this paper.
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Formanová, N., Li, XQ., Ferrie, A.M.R. et al. Towards Positional Cloning in Brassica napus: Generation and Analysis of Doubled Haploid B. rapa Possessing the B. napus pol CMS and Rfp Nuclear Restorer Gene. Plant Mol Biol 61, 269–281 (2006). https://doi.org/10.1007/s11103-006-0008-9
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DOI: https://doi.org/10.1007/s11103-006-0008-9