Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Selection of 5-Methyltryptophan and S-(2-Aminoethyl)-L-Cysteine Resistant Microspore-Derived Rice Cell Lines Irradiated with Gamma Rays

  • Kim, Dong-Sub (Mutaion Breeding Using Radiation, Korea Atomic Energy Research Institute) ;
  • Lee, In-Sok (Mutaion Breeding Using Radiation, Korea Atomic Energy Research Institute) ;
  • Jang, Cheol-Seong (Division of Biotechnology and Genetic Engineering, College of Life & Environmental Sciences, Korea University) ;
  • Hyun, Do-Yoon (Mutaion Breeding Using Radiation, Korea Atomic Energy Research Institute) ;
  • Lee, Sang-Jae (Mutaion Breeding Using Radiation, Korea Atomic Energy Research Institute) ;
  • Seo, Yong-Weon (Division of Biotechnology and Genetic Engineering, College of Life & Environmental Sciences, Korea University) ;
  • Lee, Young-Il (Mutaion Breeding Using Radiation, Korea Atomic Energy Research Institute)
  • Published : 2003.03.01
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Abstract

Microspore-derived cell lines resistant to 5-methyltryptophan (5MT, a tryptophan analog) or S-(2-aminoethyl)-L-cysteine (AEC, a Iysine analog) were selected in rice by in vitro mutagenesis. For selection of 5MT or AEC resistant cell lines, suspension-cultured cells were irradiated with gamma rays. Thirteen 5MT resistant cell lines were selected and they were able to grow stably at 2 times higher 5MT concentration. A feedback insensitive form of anthranilate synthesis, the pathway specific control enzyme for tryptophan synthesis, was detected from the 5MT resistant lines. Contents of the free amino acids in five resistant lines (MR12-1 to MR12-5) showed a 7.4 to 46.6 times greater level than that in the control culture. Tryptophan, phenylalanine, and tyrosine levels in the shikimate pathway were 28.1 and 22.5 times higher in MR12-3 and MR12 4, respectively, than that measured in the control cells. Four AEC resistant cell lines were isolated from cultures grown on medium containing 1 mM AEC, They were able to grow stably with 2 mM AEC, while sensitive calli were inhibited by 0.5 mM AEC. Aspartate kinase activities of the resistant lines were insensitive to the natural inhibitor, Iysine, and accumulated 2.2 to 12.9-fold higher levels of free Iysine than that of the control cells. Especially, the levels of aspartate, asparagine, and methionine in the aspartate pathway showed higher accumulation in the AEC resistant lines than that in the control cells.

Keywords

References

  1. Plant Cell Physiol v.37 Inheritance and stability of 5-methyltryptophan resistance in Datura innoxia selected in vitro. Brotherton JE;Schechter S;Ranch JP;Widholm JM https://doi.org/10.1093/oxfordjournals.pcp.a028958
  2. Biochem Biophys Res Commun v.41 Isolation and characterization of a lysine-sensitive aspartate kinase from a multicellular plant. Bryan PA;Cawley D;Brunner CE;Bryan JK https://doi.org/10.1016/0006-291X(70)90215-9
  3. Science v.168 Induction and isolation of auxotrophic mutants in somatic cell cultures of Nicotiana tabacum. Carlson PS https://doi.org/10.1126/science.168.3930.487
  4. Genetic manipulation with plant material In vitro selection for mutants of higher plants, In Ledoux L (ed) Chaleff RS;Calson PS
  5. Sci Sin v.18 Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Chu CC;Wang CC;Sun CS;Hsu C;Yin KC;Chu CY;Bi Fl
  6. The biochemistry of plants (vol 5. Amino acids and derivatives) Aromatic amino acid biosynthesis and its regulation. In Miflin BJ (ed) Gilchrist DG;Kosuge K
  7. Plant Cell Rep v.4 Penylalanine and tyrosine accumulating cell lines of dihaploid potato selected by resistance to 5-methyltry ptophan. Jacobsen E;Visser RGF;Wijbrandi J https://doi.org/10.1007/BF00571304
  8. In Plant mutation breeding for crop improvement v.2 New high yielding, high lysine mutants in barley. Jensen J
  9. Rice Genet Newsl v.6 Mutagenesis by means of anther culture combined with gamma irradiation. Kinoshita T;Mori K;Takamure I
  10. Theor Appl Genet v.60 Somaclonal variation-a novel source of variability from cell cultures for plant improvement. Larkin PJ;Scowcroft WR https://doi.org/10.1007/BF02342540
  11. Theor Appl Genet v.82 Selection and characterization of a rice mutant resistant to 5-methyltryptophan. Lee HY;Kameya T https://doi.org/10.1007/BF00588590
  12. Korean J Plant Tissue Culture v.19 Selection and rice cell lines resistant to 5-methyltryptophan in cell cultures. Lee HY;Kim JH;Kim HJ;Kameya T
  13. Plant Cell Tiss Org Cult v.14 Selection of regenerable maize callus cultures resistant 5-methyl-DL-tryptophan, S-2-aminoethyl-L-cysteine and high levels of L-lysine plus L-threonine. Miao S;Dunkan DR;Widholm J https://doi.org/10.1007/BF00029570
  14. Theor Appl Genet v.68 Lysine overproducer mutants with an altered dihydrodipicolinate synthase from protoplast culture of Nicotiana sylvestris (Spegazzini and Comes). Negrutiu I;Cattoir-Reynearts A;Verbruggen I;Jacobs M
  15. Plant Physiol v.71 Expression of 5-methyltryptophan resistance in plants regenerated from resistant cell lines of Datura innoxia. Ranch JP;Rick S;Brotherton JE;Widholm JM https://doi.org/10.1104/pp.71.1.136
  16. Env Exp Bot v.21 Mutations and cell selections: increased protein from regenerated rice tissue cultures. Schaeffer GW https://doi.org/10.1016/0098-8472(81)90043-5
  17. In Vitro v.17 Lysine in seed protein from Saminoethylcysteine resistant anther-derived cultures of rice. Schaeffer GW;Sharpe FT https://doi.org/10.1007/BF02618146
  18. Invited papers presented at a symposium(pp 237-254, May 16-19, 1982) Mutations and selection: Genetic variation for improved protein in rice. Genetic engineering, application to agriculture Schaeffer GW;Sharpe FT
  19. In Plant mutation breeding for crop improvement v.2 Double haploids in the mutation breeding of selected crops. Szarejko I;Maluszynski M;Polok K;Kilian A
  20. In Shively JE (ed) Methods of protein microcharacterization. Tarr GE
  21. Plant Sci v.41 Cell suspension and protoplast culture in rice. Toriyama K;Hinata K https://doi.org/10.1016/0168-9452(85)90086-X
  22. Theor Appl Genet v.74 A 5-methyltryptophan resistant rice mutant, MTR1, selected in tissue culture. Wakasa K;Widholm JM https://doi.org/10.1007/BF00290082
  23. Biochem Biophys Acta v.279 Anthranilate synthetase from 5-methyltryptophan susceptible and resistant cultured Daucus carota cells. Widholm JM https://doi.org/10.1016/0304-4165(72)90240-1
  24. Biophys Acta v.320 Measurement of th five enzymes which convert chorismate to tryptophan in cultured Daucus carota cell extracts. Widholm JM https://doi.org/10.1016/0304-4165(73)90301-2
  25. Plant Sci Lett v.3 Cultured carrot cell mutants: 5-methyltryptophan resistant trait carried from cell to plant and back. Widholm JM https://doi.org/10.1016/0304-4211(74)90003-0
  26. Can J Bot v.54 Selection and characterization of cultured carrot and tobacco cells resistant to lysine, methionine, and proline analogs. Widholm JM https://doi.org/10.1139/b76-166
  27. Crop Sci v.17 Selection and characterization of amino acid analog resistant plant cell culture. Widholm JM https://doi.org/10.2135/cropsci1977.0011183X001700040029x
  28. Current options for cereal improvement Induction of salt tolerance in highyielding rice varieties through mutagenesis and anther culture. In Maluszynski M (ed) Zapata FJ;Aldemita RR