Abstract
Analysis of reciprocal crosses between nonrestoring fertile genotypes and restored male-sterile genotypes of Lolium perenne confirmed the cytoplasmic nature of the sterility trait. This prompted a search for a molecular probe that could be used to distinguish between fertile and cytoplasmic male-sterile (CMS) cytoplasms. We describe the identification and cloning of a 4.5-kb BamHI-HindIII restriction fragment from the mtDNA of the CMS line. The cloned fragment (pCMS45) failed to hybridise to sequences in the mtDNA of fertile lines and was thus capable of unambiguously distinguishing between fertile and CMS cytoplasms. The use of pCMS45 as a diagnostic probe provided a simple test for positive identification of young non-flowering plants carrying the CMS cytoplasm and also permitted confirmation at the molecular level of the maternal transmission of the CMS trait suggested by the genetic data.
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References
Boeshore ML, Hanson MR, Izhar S (1985) A variant mitochondrial DNA arrangement specific to Petunia sterile somatic hybrids. Plant Mol Biol 4:125–132
Bonhomme S, Budar F, Ferault M, Pelletier G (1991) A 2.5 kb NcoI fragment of Ogura radish mitochondrial DNA is correlated with cytoplasmic male-sterility in Brassica cybrids. Curr Genet 19:121–127
Borck KS, Walbot V (1982) Comparison of the restriction endonuclease digestion patterns of mitochondrial DNA from normal and male-sterile cytoplasms of Zea mays L. Genetics 102:109–128
Brooks JS, Brooks MH (1967) Pollen abortion in relation to cytoplasmic genetic male sterility in sorghum. Crop Sci 7:47–51
Connolly V, Wright-Turner R (1984) Induction of cytoplasmic male sterility into ryegrass (Lolium perenne L.). Theor Appl Genet 68:449–453
Duchenne M, Lejeune B, Fouillard P, Quetier F (1989) Comparison of the organisation and expression of mtDNA of fertile and male-sterile sugar beet varieties (Beta vulgaris L.). Theor Appl Genet 78:633–664
Edwardson JR (1970) Cytoplasmic male sterility. Bot Rev 36:341–420
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Hallden C, Lind C, Sail T, Bosemark NO, Bengtsson BO (1990) Cytoplasmic male sterility in Beta is associated with structural rearrangements of the mitochondrial DNA and is not due to interspecific organelle transfer. J Mol Evol 31:365–372
Hanson MR (1991) Plant mitochondrial mutations and male sterility. Annu Rev Genet 25:461–486
Hanson MR, Conde MF (1985) Functioning and variation in cytoplasmic genomes: lessons from cytoplasmic-nuclear interactions affecting male fertility in plants. Int Rev Cytol 94:213–226
Jinks JL (1963) Cytoplasmic inheritance in fungi. In: Burdette WJ (ed) Methodology in basic genetics. Holden-Day, San Francisco, pp 325–348
Kohler RH, Horn R, Lossl A, Zetsche K (1991) Cytoplasmic male sterility in sunflower is correlated with the co-transcription of a new open reading frame with the atpA gene. Mol Gen Genet 227:369–376
Laver HK, Reynolds SJ, Moneger F, Leaver CJ (1991) Mitochondrial genome organisation and expression associated with cytoplasmic male sterility in sunflower (Helianthus annus). Plant J 1:185–193
Lee SH, Muthukrishnan S, Sorensen EL, Liang GH (1989) Restriction endonclease analysis of mitochondrial DNA from sorghum with fertile and male-sterile cytoplasms. Theor Appl Genet 77:379–382
Makaroff CA, Palmer JD (1988) Mitochondrial DNA rearrangement and transcriptional alterations in the male sterile cytoplasm of Ogura radish. Mol Cell Biol 8:1474–1480
Maniatis T, Frisch EF, Sambrook J (1982) Molecular cloning — a laboratory manual. Cold Spring Harbour Laboratory Press, Cold Spring Harbor, N.Y.
Newton KJ (1988) Plant mitochondrial genomes: organisation, expression and variation. Annu Rev Plant Physiol Plant Mol Biol 39:503–532
Siculella L, Palmer JD (1988) Physical and gene organisation of mitochondrial DNA in fertile and male sterile sunflower. CMS-associated alterations in structure and transcription of the atpA gene. Nucleic Acids Res 16:3787–3799
Small I, Suffolk R, Leaver CJ (1989) Evolution of plant mitochondrial genomes via substoichiometric intermediates. Cell 58:69–76
Umbeck PF, Gengenbach BG (1983) Reversion of male-sterile T-cytoplasm maize to male fertility in tissue culture. Crop Sci 23:584–588
Warmke HE, Lee S-LJ (1977) Mitochondrial degeneration in Texas cytoplasmic male-sterile corn anthers. J Hered 68:213–222
Weihe A, Meixner M, Wolowczyk B, Melzer R, Borner T (1991) Rapid hybridisation-based assays for the identification by DNA probes of male-sterile and male-fertile cytoplasms of the sugar beet Beta vulgaris L. Theor Appl Genet 81:819–824
Wise RP, Pring DR, Gengenbach BG (1987) Mutation to male fertility and toxin insensitivity in Texas (T)-cytoplasm maize is associated with a frameshift in a mitochondrial open reading frame. Proc Natl Acad Sci USA 84:2858–2862
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Kiang, A.S., Connolly, V., McConnell, D.J. et al. Cytoplasmic male sterility (CMS) in Lolium perenne L.: 1. Development of a diagnostic probe for the male-sterile cytoplasm. Theoret. Appl. Genetics 86, 781–787 (1993). https://doi.org/10.1007/BF00222670
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DOI: https://doi.org/10.1007/BF00222670