Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants (original) (raw)

• Journal List
• EMBO J
• v.8(3); 1989 Mar
• PMC400855

EMBO J. 1989 Mar; 8(3): 643–649.

Institute of Molecular Biology, Austrian Academy of Sciences, Billrothstrasse 11, A-5020 Salzburg, Austria

Abstract

Doubly transformed tobacco plants were obtained following sequential transformation steps using two T-DNAs encoding different selection and screening markers: T-DNA-I encoded kanamycin resistance and nopaline synthase; T-DNA-II encoded hygromycin resistance and octopine synthase. A genetic analysis of the inheritance of the selection and screening marker genes in progeny of the doubly tranformed plants revealed that the expression of T-DNA-I genes was often suppressed. This suppression could be correlated with methylation in the promoters of these genes. Surprisingly, both the methylation and inactivation of T-DNA-I genes occurred only in plants containing both T-DNAs: when self-fertilization or backcrossing produced progeny containing only T-DNA-I, expression of the genes on this T-DNA was restored and the corresponding promoters were partially or completely demethylated. These results indicated that the presence of one T-DNA could affect the state of methylation and expression of genes on a second, unlinked T-DNA in the same genome.

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• Amasino RM, Powell AL, Gordon MP. Changes in T-DNA methylation and expression are associated with phenotypic variation and plant regeneration in a crown gall tumor line. Mol Gen Genet. 1984;197(3):437–446. [PubMed] [Google Scholar]
• Ambros PF, Matzke AJM, Matzke MA. Localization of Agrobacterium rhizogenes T-DNA in plant chromosomes by in situ hybridization. EMBO J. 1986 Sep;5(9):2073–2077. [PMC free article] [PubMed] [Google Scholar]
• Chandler VL, Walbot V. DNA modification of a maize transposable element correlates with loss of activity. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1767–1771. [PMC free article] [PubMed] [Google Scholar]
• Chandler VL, Talbert LE, Raymond F. Sequence, genomic distribution and DNA modification of a Mu1 element from non-mutator maize stocks. Genetics. 1988 Aug;119(4):951–958. [PMC free article] [PubMed] [Google Scholar]
• Chomet PS, Wessler S, Dellaporta SL. Inactivation of the maize transposable element Activator (Ac) is associated with its DNA modification. EMBO J. 1987 Feb;6(2):295–302. [PMC free article] [PubMed] [Google Scholar]
• Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. [PMC free article] [PubMed] [Google Scholar]
• De Greve H, Dhaese P, Seurinck J, Lemmers M, Van Montagu M, Schell J. Nucleotide sequence and transcript map of the Agrobacterium tumefaciens Ti plasmid-encoded octopine synthase gene. J Mol Appl Genet. 1982;1(6):499–511. [PubMed] [Google Scholar]
• Delauney AJ, Tabaeizadeh Z, Verma DP. A stable bifunctional antisense transcript inhibiting gene expression in transgenic plants. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4300–4304. [PMC free article] [PubMed] [Google Scholar]
• Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM. Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet. 1982;1(6):561–573. [PubMed] [Google Scholar]
• Gelvin SB, Karcher SJ, DiRita VJ. Methylation of the T-DNA in Agrobacterium tumefaciens and in several crown gall tumors. Nucleic Acids Res. 1983 Jan 11;11(1):159–174. [PMC free article] [PubMed] [Google Scholar]
• Gritz L, Davies J. Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene. 1983 Nov;25(2-3):179–188. [PubMed] [Google Scholar]
• Hepburn AG, Clarke LE, Pearson L, White J. The role of cytosine methylation in the control of nopaline synthase gene expression in a plant tumor. J Mol Appl Genet. 1983;2(3):315–329. [PubMed] [Google Scholar]
• A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. [PubMed] [Google Scholar]
• Monk M. Genomic imprinting. Genes Dev. 1988 Aug;2(8):921–925. [PubMed] [Google Scholar]
• Otten LA, Schilperoort RA. A rapid micro scale method for the detection of lysopine and nopaline dehydrogenase activities. Biochim Biophys Acta. 1978 Dec 8;527(2):497–500. [PubMed] [Google Scholar]
• Pietrzak M, Shillito RD, Hohn T, Potrykus I. Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant expression vector. Nucleic Acids Res. 1986 Jul 25;14(14):5857–5868. [PMC free article] [PubMed] [Google Scholar]
• Potrykus I, Paszkowski J, Saul MW, Petruska J, Shillito RD. Molecular and general genetics of a hybrid foreign gene introduced into tobacco by direct gene transfer. Mol Gen Genet. 1985;199(2):169–177. [PubMed] [Google Scholar]
• St Schell J. Transgenic plants as tools to study the molecular organization of plant genes. Science. 1987 Sep 4;237(4819):1176–1183. [PubMed] [Google Scholar]
• Schernthaner JP, Matzke MA, Matzke AJ. Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants. EMBO J. 1988 May;7(5):1249–1255. [PMC free article] [PubMed] [Google Scholar]
• Watson JC, Kaufman LS, Thompson WF. Developmental regulation of cytosine methylation in the nuclear ribosomal RNA genes of Pisum sativum. J Mol Biol. 1987 Jan 5;193(1):15–26. [PubMed] [Google Scholar]
• Willmitzer L. The use of transgenic plants to study plant gene expression. Trends Genet. 1988 Jan;4(1):13–18. [PubMed] [Google Scholar]

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