Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

The increased GUS gene inactivation over generation in Arabidopsis transgenic lines

애기장대 형질전환 식물체의 세대경과에 따른 GUS유전자의 비활성화에 관한 연구

  • Park, Soon-Ki (Department of Agronomy, Kyungpook National University)
  • Published : 2002.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

The effect of transgene inactivation in T2, T3 and F2 generations was analyzed in progeny seedlings which had been generated by Agrobacterium (LBA4404/pBI121)-mediated transformation in Arabidopsis thaliana. In a system which investigated in the expression of $\beta$-glucuronidase(GUS)gene in kanamycin-resistant (ke $n^{R}$)seedlings, GUS inactivated seedlings were observed in 5 of 12 tested lines of T2 generation and the frequency of GUS inactivation was approximately 2.3%. Lines with multi-copies of T-DNA exhibited severe GUS gene inactivation with the frequency of 5.8% in T2 generation. In T3 generation lines exhibited GUS gene inactivation with the frequency of 1.3%. In contrast, inactivation increased dramatically up to 12.6% in multi-copy T-DNA line. A similar phenomenon was also found in F2 progeny from a transgenic line which had been crossed with wild-type Arabidopsis plant, WS-O (GUS gene inactivation frequency 9.9%). These results indicate that the foreign gene introduced into the plant was inactivated progressively in its transmission during subsequent generations and the transgenic line with multi-copies of T-DNA tended to show more increased inactivation.

Agrobacterium(LBA4404/pBI1121)을 이용하여 형질전환된 애기장대 (Arabidopsis thaliana)를 대상으로 T2, T3, F3세대에서의 도입된 외래 유전자의 비활성화 현상을 조사하였다. Kanamaycin저항성 개체들의 GUS유전자 발현을 분석한 결과, T2세대에서 조사된 12계통 중 5계통에서 GUS 비활성 개체가 관찰되었다 (GUS유전자 비활성율 2.3%). Multi copy T-DNA 계통을 조사한 결과, GUS 비활성 정도가 더욱 심해짐이 관찰되었다 (5.8%). T3 세대에서 single copy T-DNA 계통들은 1.3%의 GUS 비활성율을 보인 반면, multi-copy T-DNA 계통에서의 비활성율은 12.6%로 급격히 증가하였다. 유사한 현상이 형질전환 식물체와 정상개체를 교배하여 생산된 F2 계통에서도 관찰되었다 (비활성율 9.9%). 본 실험으로 식물체에 도입된 외래 유전자가 후대에서의 전이과정동안 점진적으로 비활성화되고, 이 현상은 multi copy T-DNA 계통에서 훨씬 심각함이 밝혀졌다.

Keywords

References

  1. Plant Physiol. v.99 DNA methylation is involved in maintenance of an unusual expression pattern of an introduced gene Bochardt, A.;L. Hodal;G. Palmgren;O. Mattsson;F. T. Okkels https://doi.org/10.1104/pp.99.2.409
  2. Plant J. v.5 Stable genetic transformation of Arabidopsis thaliana by Agroacterium inoculation in planta Chang, S. S.;S. K. Park;B. C. Kim;B. J. Kang;D. U. Kim;H. G. Nam https://doi.org/10.1046/j.1365-313X.1994.5040551.x
  3. Plant Mol. Biol. Rep. v.1 A plant minipreparation; version Ⅱ Dellaporta, S. L.;J. Wood;J. B. Hicks https://doi.org/10.1007/BF02712670
  4. Bio/technology v.12 Transgene inactivation;Plant fight back Finnegan, J.;D. McElroy https://doi.org/10.1038/nbt0994-883
  5. Plant Mol. Biol. v.15 The effect of T-DNA copy number, position and methylation on reporter gene expression in tobacco transformants Hobbs, S. L. A.;P. Kpodar;C. M. O. DeLong https://doi.org/10.1007/BF00039425
  6. Plant Mol. Biol. v.21 Transgene copy number can be positively or negatively associated with transgene expression Hobbs, S. L. A.;T. D. Warkentin;C. M. O. DeLong https://doi.org/10.1007/BF00039614
  7. Plant Mol. Biol. Rep. v.5 Assaying chimeric gene in plants;the GUS gene fusion system Jefferson, R. A. https://doi.org/10.1007/BF02667740
  8. EMBO J. v.6 GUS fusions;β-glucuronidase as a sensitive and versatile gene fusion marker in higher plant Jefferson, R. A.;T. A. Kavanagh;M. W. Bevan
  9. Trends Biotechnol v.8 Altered gene expression in plants due to trans interaction between homologous genes Jorgensen, R. https://doi.org/10.1016/0167-7799(90)90220-R
  10. Plant Mol. Biol. v.12 Rapid induction of genomic demethylation and T-DNA gene expression in plant cells by 5-azacytosine derivatives Klass, M.;M. C. John;D. N. Crowell;R. M. Amasino https://doi.org/10.1007/BF00017581
  11. Curr. Opinion Biotechnol. v.4 Transinactivation of gene expression in plants Kooter, J. M.;J. N. M. Mol. https://doi.org/10.1016/0958-1669(93)90118-G
  12. Mol. Gen. Genet. v.244 Homology-de-pendent gene silencing in transgenic plants;epistatic silencing loci contain multiple copies of methylated transgenes Matzke, A. J. M.;F. Neuhuber;Y. -D. Park;P. F. Ambros;M. A. Matzke
  13. EMBO J. v.8 Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants Matzke, M. A.;M. Priming;J. Trnovsky;A. J. M. Matzke
  14. Mol. Gen. Genet. v.243 Epigenetic variants of a transgenic petunia line show hypermethylation in transgene DNA;an indication for specific recognition of foreign DNA in transgene plants Meyer, P;I. Heidmann
  15. Mol. Gen. Genet. v.222 Epigenetic changes in the expression of the maize A1 gene in petunia hybrida;role of numbers of integrated gene copies and state of methylation Linn, F.;I. Heidmann;H. Saedler;P. Meyer https://doi.org/10.1007/BF00633837
  16. Plant Sci. v.88 Differential methylation and expression of the β-glucuronidase and neomycin phosphotransferase genes in transgenic plants of potato cv. Bintje Ottaviani, M-P.;T. Smits;C. H. Hanisch ten Cate https://doi.org/10.1016/0168-9452(93)90111-C
  17. Plant Mol. Biol. v.21 Treatment of Agrobacterium or leaf disks with 5-azacytidine increases transgene expression in tobacco Palmgren, G.;O. Mattson;F. T. Okkels https://doi.org/10.1007/BF00028801
  18. Mol. Cells v.3 Facile transformation of root explants of Arabidopsis thaliana L. Heynh. with a direct shooting medium Park, S. K.;B. C. Kim;S. S. Chang;H. J. Choi;S. Y. Lee;H. G. Nam;D. U. Kim
  19. Plant J. v.2 The methylation patterns of chromosomal integration regions influence gene activity of transferred DNA in petunia hybrida Prols, F;P. Meyer
  20. Plant Mol. Biol. v.25 Structure and function of selectable and non-selectable transgenes in maize after introduction by particle bombardment Register Ⅲ, J. C.;D. J. Peterson;P. J. Bell;W. P. Bullock;I. J. Evans;B. Frame;A. J. Greenland;N. S. Higgs;I. Jepson;S. Jiao;C. J. Lewnau;J. M. Sillick;H. M. Wilson https://doi.org/10.1007/BF00014669
  21. Mol. Gen. Genet. v.233 Petunia plants escapes from negative selection against a transgene by silencing the foreign DNA via methylation Penckens, S.;H. de Greve;M. van Montagu;J-P. Hernalsteens https://doi.org/10.1007/BF00587561
  22. Plant Mol. Biol. v.18 The importance of DNA methylation for stability of foreign DNA in barley Rogers, S. W.;J. C. Rogers https://doi.org/10.1007/BF00019208
  23. Mol. Gen. Genet. v.228 Reversible inactivation of a transgene in Arabidopsis thaliana Scheid, O. M.;J. Paszkowski;I. Potrykus