Abstract
Frequencies, timing, and spatial patterns of co-suppression of the nitrate (Nia) and nitrite (Nii) genes were analyzed in transgenic tobacco (Nicotiana tabacum) plants carrying either Nia or Nii cDNAs under the control of the 35S promoter, or a Nii gene with its own regulatory signals (promoter, introns, and terminator) cloned downstream of two copies of the enhancer of the 35S promoter. We show that (a) the frequencies of transgenic lines affected by co- suppression are similar for the three constructs, ranging from 19 to 25%; (b) Nia and Nii co-suppression are triggered stochastically during a phenocritical period of 2 weeks between germination and flowering; (c) the timing of co-suppression (i.e. the percentage of isogenic plants affected by co-suppression reported as a function of the number of days of culture) differs from one transgenic line to another; (d) the percentage of isogenic plants affected by co-suppression is increased by growing the plants in vitro prior to their transfer to the greenhouse and to the field; and (e) at the end of the culture period, plants are either unaffected, completely co-suppressed, or variegated. Suppressed and nonsuppressed parts of these variegated plants are separated by a vertical plane through the stem in Nia co-suppression, and separated by a horizontal plane in Nii co-suppression.
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- Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boerjan W., Bauw G., Van Montagu M., Inzé D. Distinct phenotypes generated by overexpression and suppression of S-adenosyl-L-methionine synthetase reveal developmental patterns of gene silencing in tobacco. Plant Cell. 1994 Oct;6(10):1401–1414. doi: 10.1105/tpc.6.10.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dehio C., Schell J. Identification of plant genetic loci involved in a posttranscriptional mechanism for meiotically reversible transgene silencing. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5538–5542. doi: 10.1073/pnas.91.12.5538. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorlhac de Borne F., Vincentz M., Chupeau Y., Vaucheret H. Co-suppression of nitrate reductase host genes and transgenes in transgenic tobacco plants. Mol Gen Genet. 1994 Jun 15;243(6):613–621. doi: 10.1007/BF00279570. [DOI] [PubMed] [Google Scholar]
- Dougherty W. G., Parks T. D. Transgenes and gene suppression: telling us something new? Curr Opin Cell Biol. 1995 Jun;7(3):399–405. doi: 10.1016/0955-0674(95)80096-4. [DOI] [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. doi: 10.1016/0378-1119(83)90223-8. [DOI] [PubMed] [Google Scholar]
- Hart C. M., Fischer B., Neuhaus J. M., Meins F., Jr Regulated inactivation of homologous gene expression in transgenic Nicotiana sylvestris plants containing a defense-related tobacco chitinase gene. Mol Gen Genet. 1992 Nov;235(2-3):179–188. doi: 10.1007/BF00279359. [DOI] [PubMed] [Google Scholar]
- Ingelbrecht I., Van Houdt H., Van Montagu M., Depicker A. Posttranscriptional silencing of reporter transgenes in tobacco correlates with DNA methylation. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10502–10506. doi: 10.1073/pnas.91.22.10502. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kronenberger J., Lepingle A., Caboche M., Vaucheret H. Cloning and expression of distinct nitrite reductases in tobacco leaves and roots. Mol Gen Genet. 1993 Jan;236(2-3):203–208. doi: 10.1007/BF00277113. [DOI] [PubMed] [Google Scholar]
- Linn F., Heidmann I., Saedler H., Meyer P. Epigenetic changes in the expression of the maize A1 gene in Petunia hybrida: role of numbers of integrated gene copies and state of methylation. Mol Gen Genet. 1990 Jul;222(2-3):329–336. doi: 10.1007/BF00633837. [DOI] [PubMed] [Google Scholar]
- Matzke M. A., Matzke AJM. How and Why Do Plants Inactivate Homologous (Trans)genes? Plant Physiol. 1995 Mar;107(3):679–685. doi: 10.1104/pp.107.3.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matzke M. A., Neuhuber F., Matzke A. J. A variety of epistatic interactions can occur between partially homologous transgene loci brought together by sexual crossing. Mol Gen Genet. 1993 Jan;236(2-3):379–386. doi: 10.1007/BF00277137. [DOI] [PubMed] [Google Scholar]
- Matzke M. A., Primig M., Trnovsky J., Matzke A. J. Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants. EMBO J. 1989 Mar;8(3):643–649. doi: 10.1002/j.1460-2075.1989.tb03421.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyer P., Heidmann I., Niedenhof I. Differences in DNA-methylation are associated with a paramutation phenomenon in transgenic petunia. Plant J. 1993 Jul;4(1):89–100. doi: 10.1046/j.1365-313x.1993.04010089.x. [DOI] [PubMed] [Google Scholar]
- Palauqui J. C., Vaucheret H. Field trial analysis of nitrate reductase co-suppression: a comparative study of 38 combinations of transgene loci. Plant Mol Biol. 1995 Oct;29(1):149–159. doi: 10.1007/BF00019126. [DOI] [PubMed] [Google Scholar]
- Saux C., Lemoine Y., Marion-Poll A., Valadier M. H., Deng M., Morot-Gaudry J. F. Consequence of Absence of Nitrate Reductase Activity on Photosynthesis in Nicotiana plumbaginifolia Plants. Plant Physiol. 1987 May;84(1):67–72. doi: 10.1104/pp.84.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaucheret H., Kronenberger J., Lepingle A., Vilaine F., Boutin J. P., Caboche M. Inhibition of tobacco nitrite reductase activity by expression of antisense RNA. Plant J. 1992 Jul;2(4):559–569. [PubMed] [Google Scholar]
- Vaucheret H., Palauqui J. C., Elmayan T., Moffatt B. Molecular and genetic analysis of nitrite reductase co-suppression in transgenic tobacco plants. Mol Gen Genet. 1995 Aug 21;248(3):311–317. doi: 10.1007/BF02191598. [DOI] [PubMed] [Google Scholar]
- Vincentz M., Caboche M. Constitutive expression of nitrate reductase allows normal growth and development of Nicotiana plumbaginifolia plants. EMBO J. 1991 May;10(5):1027–1035. doi: 10.1002/j.1460-2075.1991.tb08041.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vincentz M., Moureaux T., Leydecker M. T., Vaucheret H., Caboche M. Regulation of nitrate and nitrite reductase expression in Nicotiana plumbaginifolia leaves by nitrogen and carbon metabolites. Plant J. 1993 Feb;3(2):315–324. doi: 10.1111/j.1365-313x.1993.tb00183.x. [DOI] [PubMed] [Google Scholar]
- de Carvalho Niebel F., Frendo P., Van Montagu M., Cornelissen M. Post-transcriptional cosuppression of beta-1,3-glucanase genes does not affect accumulation of transgene nuclear mRNA. Plant Cell. 1995 Mar;7(3):347–358. doi: 10.1105/tpc.7.3.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Carvalho F., Gheysen G., Kushnir S., Van Montagu M., Inzé D., Castresana C. Suppression of beta-1,3-glucanase transgene expression in homozygous plants. EMBO J. 1992 Jul;11(7):2595–2602. doi: 10.1002/j.1460-2075.1992.tb05324.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van der Krol A. R., Mur L. A., Beld M., Mol J. N., Stuitje A. R. Flavonoid genes in petunia: addition of a limited number of gene copies may lead to a suppression of gene expression. Plant Cell. 1990 Apr;2(4):291–299. doi: 10.1105/tpc.2.4.291. [DOI] [PMC free article] [PubMed] [Google Scholar]