Molecular Cloning of Putrescine N-Methyltransferase Gene from Burley 21 Tobacco

Burley 21 담배에서 Putrescine N-Methyltransferase 유전자의 클로닝

  • Published : 2003.12.01

Abstract

Recently, many researches for plant alkaloids, one of the largest groups of natural products, are reported because of their various pharmacological activity. This study was carried out to clone putrescine N-methyltransferase (PMT) gene which is a key enzyme in diverting polyamine metabolism towards the biosynthesis of nicotine and related alkaloids from Burley tobacco. To induce expression of PMT gene in tobacco plant, the floral meristem was removed and then mRNA was purified from root. cDNA encoding PMT gene was isolated by RT PCR and cloned. Three different groups of clones were screened by PCR and restriction enzyme digestion analysis and were characterized. The data of these screening revealed that three types of PMT are present in Burley tobacco. Comparison of the nucleotide sequence of this three genes encoding putative PMT with those of other tobaccos revealed that two types of PMT are newly discovered from Nicotiana tabacum cv. Br21 tobacco and they were same as PMT2, PMT3 of N. tabacum cv. Xanthi.

Keywords

References

  1. De Luca, V. (1993) Enzymology of indole alkaloid biosynthesis. In Method in Plant Biochemistry, ed. PM Dey, JB Harbome, London. Academic 9: 345-368
  2. Fumihiko, S., Takashi, H., Akira, H., Kenichi, T., Choi, K.B., Takashi, M., Hideki, F. and Yasuyuki, Y. (2001) Metabolic engineering of plant alkaloid biosynthesis Proc. Natl. Acad. Sci. Vol98, No.1:367-372 https://doi.org/10.1073/pnas.011526398
  3. Hashimoto, T. and Yamada, Y. (1992) Tropane alkaloid biosynthesis : regulation and application. In Pro. 7th Annu. Ponn, state Symp. Plant Physiol. Rockville : Amm. Soc. Plant PhysiI. Press. 122-134
  4. Hashimoto, T. and Yamata, Y. (1993) Nicotine and tropane alkaloids. J. Plant Res. Special Issue. 3 : 369-379
  5. Hibi, N. (I994) Ph D. Thesis, Biochemical and molecular analysis of putrescine N-methyltransferase in plants. Kyoto university, Kyoto, Japan
  6. Hibi, N., Higashiguchi, S., Hashimoto, T. and Yamada, Y. (1994) Gene expression in tobacco low nicotine mutants. Plant Cell 6: 723-735 https://doi.org/10.1105/tpc.6.5.723
  7. Kinnersley, A. M. and Dougall, D. K. (1980) Correlation between the nicotine content of tobacco plants and callus cultures. Planta 149: 205-206 https://doi.org/10.1007/BF00380884
  8. Riechers, D. E. and Timko, M. P. (1999) Structure and expression of the gene family encoding putrescine N-methyltransferase in Nicotiana tabacum: new clues to the evolutionary origin of cultivated tobacco. Plant Mol. Biol. 41(3):387-401 https://doi.org/10.1023/A:1006342018991
  9. Saitoh, F., Noma, M. and Kawashima, N. (I985) The alkaloid contents of sixty Nicotiana species. Phytochemistry 24: 477-480 https://doi.org/10.1016/S0031-9422(00)80751-7
  10. Tiburcio, A. F., Ingersoll, R. and Galston, A. W. (1985) Modified alkaloid pattern developing tobacco callus. Plant Sci. 38: 207-212 https://doi.org/10.1016/0168-9452(85)90040-8
  11. Tiburcio Pinol M. T. and Serrano M. (1985) Effect of UV-C on growth, soluble protein and alkaloids in Nicotiana rustica plas. Environ. Exp. Bot. 25: 203-210 https://doi.org/10.1016/0098-8472(85)90004-8
  12. Winz, R. A. and Baldwin, I. T. (2001) Molecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuaia. IV. Insect-Induced ethylene reduces jasmonate-induced nicotine accumulation by regulating putrescine N-methyltransferase transcripts. Plant Physiol: 125(4): 2189-2202 https://doi.org/10.1104/pp.125.4.2189
  13. 이상하, 민영근 (1987) 담배과학 총설, 한국연초학회, 제일문화사, 한국
  14. 한국인삼연초연구소 (1983) 담배품종도감, 한국인삼연초연구소, 고려서적주식회사, 한국