Abstract
Melandrium album (syn.Silene latifolia) is a model dioecious species in which theY chromosome, present only in heterogametic males, plays both a male-determining and a strict female-suppressing role. We showed that treatment with 5-azacytidine (5-azaC) induces a sex change to androhermaphroditism (andromonoecy) in about 21% of male plants, while no apparent phenotypic effect was observed in females. All of these bisexual androhermaphrodites (with the standard male 24,AA +XY karyotype) were mosaics possessing both male and hermaphrodite flowers and, moreover, the hermaphrodite flowers displayed various degrees of gynoecium development and seed setting. Southern hybridization analysis with a repetitive DNA probe showed that the 5-azacytidine-treated plants were significantly hypomethylated in CG doublets, but only to a minor degree in CNG triplets. The bisexual trait was transmitted to two successive generations, but only when androhermaphrodite plants were used as pollen donors. The sex reversal was inherited with incomplete penetrance and varying expressivity. Based on the uniparental inheritance pattern of androhermaphroditism we conclude that it originated either by 5-azaC induced inhibition ofY-linked female-suppressing genes or by a heritable activation of autosomal female-determining/promoting genes which can be reversed, on passage through female meiosis, by a genomic imprinting mechanism. The data presented indicate that female sex suppression inM. album XY males is dependent on methylation of specific DNA sequences and can be heritably modified by hypomethylating drugs.
Similar content being viewed by others
References
Amasino RM, Powell ALT, Gordon MP (1984) Changes in T-DNA methylation and expression are associated with phenotypic variation and plant regeneration in a crown gall tumor line. Mol Gen Genet 197:437–446
Bezděk M, Koukalová B, Brzobohatý B, Vyskot B (1992) Differential sensitivity of CG and CCG DNA sequences to ethionine-induced hypomethylation of theNicotiana tabacum genome. FEBS Lett 300:268–270
Burn JE, Bagnall DJ, Metzger JD, Dennis ES, Peacock WJ (1993) DNA methylation, vernalization, and the initiation of flowering. Proc Natl Acad Sci USA 90:287–291
Cedar H, Solage A, Glaser G, Razin A (1979) Direct detection of methylated cytosine in DNA by use of the restriction enzymeMspI. Nucleic Acids Res 6:2125–2132
Chaillet JR, Vogt TF, Beier DR, Leder P (1991) Parental-specific methylation of an imprinted transgene is established during gametogenesis and progressively changes during embryogenesis. Cell 66:77–83
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Reporter 1:19–21
Emlen JM, Freeman DC, Graham JH (1993) Non-linear growth dynamics and the origin of fluctuating asymmetry. Genetica 89:77–96
Fajkus J, Reich J (1991) Evaluation of restriction endonuclease cleavage of plant nuclear DNA using contaminating chloroplast DNA. Folia Biol (Prague) 37:224–226
Flavell RB (1986) The structure and control of expression of ribosomal RNA genes. Oxford Surv Plant Mol Cell Biol 3:251–274
Grant S, Houben A, Vyskot B, Široký J, Wei-Hua P, Macas J, Saedler H (1994) Genetics of sex determination in flowering plants. Dev Genet 15:214–230
Gruenbaum Y, Naveh-Many T, Cedar H, Razin A (1981) Sequence specifity of methylation in higher plant DNA. Nature 292:860–862
Holliday R (1990) DNA methylation and epigenetic inheritance. Phil Trans Royal Soc London B326:329–338
Jones PA (1985) Altering gene expression with 5-azacytidine. Cell 40:485–486
Kermicle JL, Alleman M (1990) Gametic imprinting in maize in relation to the angiosperm life cycle. Development 1990 (Suppl): 9–14
Kiss T, Kis M, Solomosy F (1989) Nucleotide sequence of a 25S rRNA gene from tomato. Nucleic Acids Res 17:796
Matzke M, Matzke AJM (1993) Genomic imprinting in plants: parental effects andtrans-inactivation phenomena. Annu Rev Plant Physiol Plant Mol Biol 44:53–76
Meyer P, Heidmann I, Niedenhof I (1993) Differences in DNA methylation are associated with a paramutation phenomenon in transgenic petunia. Plant J 4:89–100
Meyer P, Niedenhof I, Lohuis MT (1994) Evidence for cytosine methylation of non-symmetrical sequences in transgenicPetunia hybrida. EMBO J 13:2084–2088
Pradhan S, Adams RLP (1995) Distinct CG and CNG DNA methyltransferases inPisum sativum. Plant J 7:471–481
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual (2nd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sano H, Kamada I, Youssefian S, Katsumi M, Wabiko H (1990) A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol Gen Genet 220:441–447
Schwartz D, Dennis E (1986) Transposase activity of theAc controlling element in maize is regulated by its degree of methylation. Mol Gen Genet 205:476–482
Široký J, Janoušek B, Mouras A, Vyskot B (1994) Replication pattern of sex chromosomes inMelandrium album female cells. Hereditas 120:175–181
Sugiura M, Shinozaki K, Zaita N, Kusuda M, Kumano M (1986) Clone bank of the tobacco (Nicotiana tabacum) chloroplast genome as a set of overlapping restriction endonuclease fragments: mapping of eleven ribosomal protein genes. Plant Sci 44:211–216
Surani MA, Reik W, Allen ND (1988) Transgenes as molecular probes for genomic imprinting. Trends Genet 4:59–62
Tamame M, Antequera F, Santos E (1988) Developmental characterization and chromosomal mapping of the 5-azacytidine-sensitivefluF locus ofAspergillus nidulans. Mol Cell Biol 8:3043–3050
Vagera J, Paulíková D, Doležel J (1994) The development of male and female regenerants byin vitro androgenesis in the dioecious plantMelandrium album. Ann Bot 73:455–459
Van Nigtevecht G (1966) Genetic studies in dioeciousMelandrium. II. Sex determination inMelandrium album andMelandrium dioicum. Genetica 37:307–344
Vyskot B, Araya A, Veuskens J, Negrutiu I, Mouras A (1993) DNA methylation of sex chromosomes in a dioecious plant,Melandrium album. Mol Gen Genet 239:219–224
Vyskot B, Koukalová B, Kovařík A, Sachambula L, Reynolds D, Bezděk M (1995) Meiotic transmission of a hypomethylated repetitive DNA family in tobacco. Theor Appl Genet 91:659–664
Westergaard M (1948) The relation between chromosome constitution and sex in the offspring of triploidMelandrium. Hereditas 34:257–279
Winge O (1931) X and Y linked inheritance inMelandrium. Hereditas 15:127–165
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
Janoušek, B., Široký, J. & Vyskot, B. Epigenetic control of sexual phenotype in a dioecious plant,Melandrium album . Molec. Gen. Genet. 250, 483–490 (1996). https://doi.org/10.1007/BF02174037
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02174037