Abstract
DNA molecules from mitochondria of whole plants and a suspension culture ofChenopodium album were prepared, by a gentle method, for analysis by electron microscopy. Mitochondrial (mt) DNA preparations from both sources contained mostly linear molecules of variable sizes (with the majority of molecules ranging from 40 to 160 kb). Open circular molecules with contour lengths corresponding to 0.3–183 kb represented 23–26% of all mtDNA molecules in the preparations from the suspension culture and 13–15% in the preparations from whole plants. More than 90% of the circular DNA was smaller than 30 kb. Virtually no size classes of the mtDNA molecules could be identified, and circular or linear molecules of the genome size (about 270 kb) were not observed. In contrast, plastid (pt) DNA preparations from the suspension culture contained linear and circular molecules falling into size classes corresponding to monomers, dimers and trimers of the chromosome. About 23% of the ptDNA molecules were circular. DNA preparations from mitochondria contained a higher percentage of more complex molecules (rosette-like structures, catenate-like molecules) than preparations of ptDNA. Sigma-like molecules (putative intermediates of rollingcircle replication) were observed in mtDNA preparations from the suspension culture (18% of the circles), and in much lower amount (1%) in preparations from whole plants. The results are compared with data obtained previously by pulsed-field gel electrophoresis and discussed in relation to the structural organization and replication of the mt genome of higher plants.
Similar content being viewed by others
References
André CP, Walbot V (1995) Pulsed-field gel mapping of maize mitochondrial chromosomes. Mol Gen Genet 247:255–263
André CP, Levy AA, Walbot V (1992) Small repeated sequences and the structure of plant mitochondria genomes. Trends Genet 8:128–132
Backert S, Dörfel P, Börner T (1995) Investigation of plant organellar DNA by pulsed-field gel electrophoresis. Curr Genet 28:390–399
Bailey-Serres J, Leroy P, Jones SS, Wahleithner JA, Wolstenholme DR (1987) Size distributions of circular molecules in plant mitochondrial DNAs. Curr Genet. 12:49–53
Bendich AJ (1993) Reaching for the ring: the study of mitochondrial genome structure. Curr Genet 24:279–290
Bendich AJ, Smith SB (1990) Moving pictures and pulsed-field gel electrophoresis show linear DNA molecules from chloroplasts and mitochondria. Curr Genet 17:421–425
Bendich AJ, Loretz CJ, Monnat RJ Jr (1993) The structure of the plant mitochondrial genome. In: Brennicke A, Kück U (eds) Plant mitochondria. Verlag Chemie, Weinheim, pp 171–180
Beverley SM (1988) Characterization of the ‘unusual’ mobility of large circular DNAs in pulsed field-gradient electrophoresis. Nucleic Acids Res 16:926–939
Bohnert HJ (1977) Size and structure of mitochondrial DNA fromPhysarum polycephalum. Exp Cell Res 106:426–430
Brennicke A, Blanz P (1982) Circular mitochondrial DNA species fromOenothera with unique sequences. Mol Gen Genet 187:461–466
Burkhardt H, Lurz R (1982) Electron microscopy. In: Pühler A, Timmis KN (eds) Advanced molecular genetics. Springer-Verlag, Berlin, pp 281–308
Dale RMK, Duesing JH, Keene D (1981) Supercoiled mitochondrial DNAs from plant tissue-culture cells. Nucleic Acids Res 9:4583–4593
Dinouel N, Drissi R, Miyakawa I, Sor F, Rousset S, Fukuhara H (1993) Linear mitochondrial DNAs of yeasts: closed-loop structure of the termini and possible linear-circular conversion mechanisms. Mol Cell Biol 13:2315–2323
Dörfel P, Weihe A, Knösche R, Börner T (1989) Mitochondrial DNA ofChenopodium album (L.): comparison of leaves and suspension cultures. Curr Genet 16:375–380
Dörfel P, Weihe R, Dolferus R, Börner T (1991) DNA sequence of a mitochondrial plasmid fromChenopodium album. Plant Mol Biol 17:155–156
Dudareva NA, Kiseleva EV, Boyarintseva AG, Maystrenko AG, Khrystolyubova NB, Sagalnik RI (1988) Structure of the mitochondrial genome ofBeta vulgaris L. Theor Appl Genet 76:753–759
Fauron CMR, Havlik M. Bretell RIS (1990) The mitochondrial genome organization of a maize fertile cmsT revertant is generated through recombination between two sets of repeats. Genetics 124:423–428
Fontarnau A, Hernández-Yago J (1982) Characterization of mitochondrial DNA inCitrus. Plant Physiol 70:1678–1682
Gilbert W, Dressler D (1968) DNA replication the rolling-circle model. Cold Spring Harbor Symp Quant Biol 33:473–484
Goddard JM, Cummings DJ (1975) Structure and replication of mitochondrial DNA fromParamecium aurelia. J Mol Biol 97:593–609
Gray MW (1989) Origin and evolution of mitochondrial DNA. Annu Rev Cell Biol 5:25–50
Gruss A, Ehrlich SD (1989) The family of highly interrelated singlestranded DNA plasmids. Microbiol Rev 53:231–241
Han ZY, Stachow C (1994) Analysis ofSchizosaccharomyces pombe mitochondrial DNA replication by two-dimensional gel electrophoresis. Chromosoma 103:162–170
Handa H, Tsunewaki K, Kumisada T, Yamagashi H (1984) Small circular DNA molecules in wheat mitochondria. Mol Gen Genet 194:368–372
Kenwrick S, Pattersohn M, Speer A, Fischbeck K, Davies K (1987) Molecular analysis of the Duchenne muscular dystrophy region using pulsed-field gel electrophoresis. Cell 48:351–357
Kim BD, Lee KJ, de Busk AG (1982) Linear and ‘lasso-like’ structures of mitochondrial DNA fromPennisetum typhoides. FEBS Lett 147:231–234
Knösche R, Günther G (1988) A cell-division cycle in suspension cultures fromChenopodium album with unspecific arrest at G1 and G2 phase under stationary growth conditions. Biol Zentralbl 107:653–661
Kolodner P, Tewari KK (1972) Physicochemical characterization of mitochondrial DNA from pea leaves. Proc Natl Acad Sci USA 69:1830–1834
Kool AJ, de Haas JM, Mol JNM, van Marrewijk GAM (1985) Isolation and physicochemical characterization of mitochondrial DNA from cultured cells ofPetunia hybrida. Theor Appl Genet 69:223–233
Koths K, Dressler D (1978) Analysis of ϕXDNA replication cycle by electron microscopy. Proc Natl Acad Sci USA 75:605–609
Lang D, Mitani M (1970) Simplified quantitative electron microscopy of biopolymers. Biopolymers 9:373–379
León P, Macaya G (1983) Properties of DNA-rosettes and their relevance to chromosome structure. Chromosoma 88:307–314
Levenne SD, Zimm BH (1987) Separation of open-circular DNA using pulsed-field electrophoresis. Proc Natl Acad Sci USA 84:4054–4057
Levings CS III, Shah DM, Hu WWL, Pring DR, Timothy DH (1979) Molecular heterogeneity among mitochondrial DNAs from different maize cytoplasms. In: Cummings DJ, Borst P, David IB, Weissman SM, Fox C (eds) Extrachromosomal DNA. ICN-UCLA Symp Mol Cell Biol 15. Academic Press, New York, pp 63–73
Levy AA, André CP, Walbot V (1991) Analysis of a 120-kilobase mitochodrial chromosome in maize. Genetics 128:417–424
Lonsdale DM, Hodge TP, Fauron CM-R (1984) The physical map and organization of the mitochondrial genome from the fertile cytoplasm of maize. Nucleic Acids Res 12:9249–9261
Lonsdale DM, Brears T, Hodge TP, Melville SA, Rottman WH (1988) The plant mitochondrial genome: homologous recombination as a mechanism for generating heterogeneity. Phil Trans R Soc Lond B 319:140–163
Maleszka R, Skelly PJ, Clark-Walker GD (1991) Rolling-circle replication of DNA in yeast mitochondria. EMBO J 10:3923–3929
Manna F, Del Giudice L, Massardo DR, Schreil WH, Cermola M, Devreux M, Wolf K (1985) The mitochondrial genome ofNicotiana plumbaginifolia. Curr Genet 9:411–415
Negruk VI, Eisner GI, Redichkina TD, Dumanskaya NN, Cherny DI, Alexandrov AA, Shemyakin MF, Butenko RG (1986) Diversity ofVicia faba circular mtDNA in whole plants and suspension cultures. Theor Appl Genet 72:541–547
Oda K, Yamato K, Ohta E, Nakamura Y, Takemura M, Nozato N, Akashi K, Kanegae T, Ogura Y, Kohchi T, Ohyama K (1992) Gene organization deduced from the complete sequence of liverwortMarchantia polymorpha mitochondrial DNA. J Mol Biol 223:1–7
Narayanan KK, André CP, Yang J, Walbot V (1993) Organization of a 117-kb circular mitochondrial chromosome in IR36 rice. Curr Genet 23:248–254
Palmer JD (1992) Chloroplast and mitochondrial genome evolution in land plants. In: Herrmann RG (ed) Cell organelles. Springer-Verlag, Wien, pp 99–133
Palmer JD, Shields CR (1984) Tripartite structure of theBrassica compestris mitochondrial genome. Nature 307:437–440
Schardl CL, Lonsdale DM, Pring DR, Rose KR (1984) Linearization of maize mitochondrial chromosomes by recombination with linear episomes. Nature 301:292–296
Schuster W, Brennicke A (1994) The plant mitochondrial genome: physical structure, information content, RNA editing, and gene migration to the nucleus. Annu Rev Plant Physiol Plant Mol Biol 45:61–78
Schuster W, Hiesel B, Wiesinger B, Schobel W, Brennicke A (1988) Isolation and analysis of plant mitochondria and their genomes. In: Shaw CH (ed) Plant molecular biology — a practical approach, IRL Press, Oxford, pp 79–102
Shapiro TA (1993) Kinetoplast DNA maxicircles: networks within networks. Proc Natl Acad Sci USA 90:7809–7813
Siemenroth A, Wollgiehn R, Neumann D, Börner T (1981) Synthesis of ribosomal RNA in ribosome-deficient plastids of the mutant ‘albostrians’ ofHordeum vulgare L Planta 153:547–555
Small ID, Suffolk R, Leaver CJ (1989) Evolution of plant mitochondrial genomes via substoichiometric intermediates. Cell 58:69–76
Solar G-del, Moscoso M, Espinosa M (1993) Rolling circle-replicating plasmids from gram-positive and gram-negative bacteria: a wall falls. Mol Microbiol 8:789–796
Sparks RB Jr, Dale RMK (1980) Characterization of3H-labeled supercoiled mitochondrial DNA from tobacco suspension culture cells. Mol Gen Genet 180:351–355
Synenki RM, Levings CS III, Shah DM (1978) Physicochemical characterization of mitochondrial DNA from soybean. Plant Physiol 61:460–464
Unseld M, Brandt P, Heinze B, Eckert-Ossenkopp U, Brennicke A (1993) The mitochondrial genome ofArabidopsis thaliana. In: Brennicke A, Kück U (eds) Plant mitochondrial. Verlag Chemie, Weinheim, pp 137–143
Wahleithner JA, Wolstenholme DR (1988) Origin and direction of replication in mitochondrial plasmid DNAs of broad bean,Vicia faba. Curr Genet 14:163–170
Wang B, Cheng W, Li Y-N, Li D-D (1989) Some physicochemical properties of rice mitochondrial DNA. Theor Appl Genet 77:581–586
Ward BL, Anderson RS, Bendich AJ (1981) The mitochondrial genome is large and variable in a family of plants (Cucurbitaceae). Curr Genet 12:55–67
Wilson AJ, Choury PS (1984) An rapid inexpensive method for the isolation of restrictable mitochondrial DNA from various plant sources. Plant Cell Rep 3:237–239
Author information
Authors and Affiliations
Additional information
Communicated by H. Kössel
Rights and permissions
About this article
Cite this article
Backert, S., Lurz, R. & Börner, T. Electron microscopic investigation of mitochondrial DNA fromChenopodium album (L.). Curr Genet 29, 427–436 (1996). https://doi.org/10.1007/BF02221510
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02221510