Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Antisense GA 3β-Hydroxylase Gene Transferred to Rice Plants.

Antisense gibberellin 3β-hydroxylase발현 형질전환벼

  • 강용원 (경북대학교 농업생명과학대학 식물생명과학부) ;
  • 윤용휘 (경북대학교 농업생명과학대학 식물생명과학) ;
  • 김길웅 (경북대학교 농업생명과학대학 식물생명과학) ;
  • 이인중 (경북대학교 농업생명과학대학 식물생명과학) ;
  • 신동현 (경북대학교 농업생명과학대학 식물생명과학부)
  • Published : 2004.08.01
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Abstract

During plant development, active gibberellins (GAs) control many aspects of plant growth and development including seed germination, stem elongation, flower induction, anther development and seed growth. To understand the biosynthesis and functional role of active GAs in high plants, this study investigated GA 3$\beta$-hydroxylase gene en-coding $GA_1$ and$GA_4$ catalizing last step in GA biosynthetic pathway. The antisense GA 3$\beta$-hydroxylase gene was inserted into expression vector, pIG121-Hm. Calli derived from mature seeds of rice (Oryza satiiva L. cv. Donjinbyeo) were co-cultivated with Agrohacterium tumefaciens EHA101 earring a pIG121-Hm containing hygromycin resistance ($Hyg^r$) and antisense GA 3$\beta$-hydroxylase gene. Seventeen transgenic plants obtained inhibiting GA 3$\beta$-hydroxylase. Transgenic plants had shorter plant height more than that of the Dongjinbyeo. Stable integration of antisense GA 3$\beta$-hydroxylase gene was confirmed by polymerase chain reaction of genomic DNA isolated from the leaf organs of the $T_o$ generation.

GA 생합성에 결정적 역할을 하는 GA 3$\beta$-hydroxylase를 pIG121-Hm 벡터에 GUS유전자를 빼고 antisense로 클로닝하여 이를 동진벼에 도입한 결과 17개체의 절간신장이 억제된 형질전환한 식물체를 얻을 수 있었다. 일반 재배 동진벼를 대조군으로 하여 비교하였을때 antisense GA 3$\beta$-hydroxylase 유전자가 형질 도입된 식물체의 획득형질은 평균적으로 대조군에 비해 절간 신장의 억제가 확인되었다. 절간신장의 억제가 보인 개체의 엽육조직을 co취하여 Southen blot hybridization분석 결과 3개의 line에서 모두 single copy로 도입된 것으로 나타났다. 이로써$T_o$ 식물체 내에 antisense GA 3$\beta$-hydroxylase 유전자를 내포하고 있는 것으로 확인되었다. 이것은 antisense GA 3$\beta$-hydroxylase 유전자가 생체내에서 직접 또는 간접적으로 GA 3$\beta$-hydroxylase유전자의 발현에 관여한 것이라 사료되어진다.

Keywords

References

  1. Badula, S. K. and V. K. Sawhaney. 1991. Protein analysis during the ontogeny of ormal and male sterile stamenless- 2 mutant stamens of tomato. Biochem Genet. 29, 29-41 https://doi.org/10.1007/BF00578237
  2. Chan, M. T., H. H. Chang., S. L. Ho., W. F. Tong and S. M. Yu. 1993. Agrobacterium-mediated production of transgenic rice plants expressing a chimeric alpha-amylase promoter/beta-glucuronidase gene. Plant Mol Biol. 22(3), 491- 506 https://doi.org/10.1007/BF00015978
  3. Chiang, H. H., I. Hwang and H. M. Goodman. 1995. Isolation of the Arabidopsis GA4 locus. Plant Cell. 7, 195-201 https://doi.org/10.1105/tpc.7.2.195
  4. Fujioka, S., H. Yamane & Spray, C. R., P. Gaskin, J. MacMillan, B. O. Phinney and N. Takahashi. 1988. Qualitative and quantitative analyses of gibberellins in vegetative shoots of normal, dwarf -1, dwarf- 2, dwarf- 3, and dwarf- 5 seedlings of Zea mays L. Plant Physiol. 88, 1367 -1372 https://doi.org/10.1104/pp.88.4.1367
  5. Hedden, P. and Y. Kamiya. 1997. Gibberellin biosynthesis: Enzymes, genes and their regulation. Annu. Rev. Physiol. Plant Mol. Biol. 48, 431-460 https://doi.org/10.1146/annurev.arplant.48.1.431
  6. Hiei, Y., T. Komari and T. Kubo. 1994. Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol Biol. 35(1-2), 205-218 https://doi.org/10.1023/A:1005847615493
  7. Ingram, T. J., J. B. Reid, W. C. Potts and I. C. Murfet. 1983. Internode length in Pisum: The Le gene controls the 3$\beta$- hydroxylation of gibberellin $A_2_0$to gibberellin $A_1$. Physiol. Plant. 59, 607-616 https://doi.org/10.1111/j.1399-3054.1983.tb06288.x
  8. Itoh, H., M. Tanaka-Ueguchi, H. Kawaide, X. Chen, Y. Kamiya and M. Matsuoka. 1999. The gene encoding tobacco gibberellin 3$\beta$-hydroxylase is expressed at the site of GA action during stem elongation and flower organ development. Plant J. 20, 15-24 https://doi.org/10.1046/j.1365-313X.1999.00568.x
  9. Jacobsen, S. E and N. E. Olszewski. 1991. Characterization of the arrest in anther development associated with gibberellin deficiency of the gib-1 mutant of tomato. Plant Physiol. 97, 409-414 https://doi.org/10.1104/pp.97.1.409
  10. Kaneko, M., H. Itoh, M. Ueguchi-Takara, M. Ashikari and M. Matruoka. 2002. The alpha-amylase induction in endosperm during rice seed germination is caused by gibberellin synthesized in epithelium. Plant Physiol. 128(4), 1264-1270 https://doi.org/10.1104/pp.010785
  11. Kobayashi, M., A. Sakurai, H. Saka and N. Takahashi. 1989. Quantitative - analysis of endogenous gibberellins in normal and dwarf cultivars of rice. Plant Cell Physiol. 30, 963-969
  12. Kobayashi, M., Y. Kamiya, A. Sakurai, H. Saka and N. Takahashi. 1990. Metabolism of gibberellins in cell-free- extracts of anthers from normal and dwarf rice. Plant Cell Physiol. 31, 289-293
  13. Kinoshita, T and N. Shinbashi. 1982. Identification of dwarf gene and their Character expressions in the isogenic back-ground Genetical studies on rice plant. Jpn. J. Breed. 32, 219-231 https://doi.org/10.1270/jsbbs1951.32.219
  14. Koornneef, M and J. H. Van der Veen. 1980. Induction and analysis of gibberellin sensitive mutants in Arabidopsis thaliana (L) Heynh. Theor. Appl. Genet. 58, 257-263 https://doi.org/10.1007/BF00265176
  15. Lange, T., S. Robatzek and A. Frisse. 1997. Cloning and expression of a gibberellin 2ß, 3$\beta$-hydroxylase cDNA from pumpkin endosperm. Plant Cell. 9, 1459-1467 https://doi.org/10.1105/tpc.9.8.1459
  16. Murakami, Y. 1972. Dwarfing genes in rice and their relation to gibberellin biosynthesis. In Carr DJ, ed, Plant Growth Substrances. p166-174
  17. Murray, M. C and W. F. Thompson. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 8, 4321-4325 https://doi.org/10.1093/nar/8.19.4321
  18. Rasid, H., S. Yokoi, K. Toriyama and K. Hinata. 1996. Trangenic plant production mediated by Agrobacterium in indica rice. Plant Cell Rep. 15, 727-730 https://doi.org/10.1007/BF00232216
  19. Ross, J. J., I. C. Murfet and J. B. Reid. 1997. Gibberellin mutants. Physiol. plant. 100, 550-560 https://doi.org/10.1111/j.1399-3054.1997.tb03060.x
  20. Saghai-Maroof, M. A., K. A. Soliman, R. A. Jorgensen and R. W. Allard. 1984. Ribosomal DNA spacerlength polymorphisms in barley : Mendelian inheritance, chromosomal location, and population dyanmics. Proc Natl Acad Sci USA. 81, 8014-8018 https://doi.org/10.1073/pnas.81.24.8014
  21. Silverstone, A. L., C.-W. Chang, E. Krol and T. P. Sun. 1997. Developmental regulation of the gibberellin biosynthetic gene GA1 in Arabidopsis thaliana. Plant J. 12, 9-19 https://doi.org/10.1046/j.1365-313X.1997.12010009.x
  22. Smith, R. H. and E. E. Hood. 1995. Reviw and interpretention : Agrobacterium tumefaciens transformation of monocotyledons. Crop Sci. 301-309
  23. Toyomasu, T., H. Kawaide, W. Mitsuhashi, Y. Inoue and Y. Kamiya. 1998. Phytochrome regulates gibberellin biosynthesis during germination of photoblastic lettuce seeds. Plant Physiol. 118, 1517-1523 https://doi.org/10.1104/pp.118.4.1517
  24. Talon, M., M. Koornneef and J. A. Zeevaart. 1990. Endofenous gibberellins in Arabidopsis thaliana and possible steps blocked in the biosynthetic pathways of the semidwarf ga4 and ga5 mutants. Proc Natl Acad Sci USA. 87(20), 7983-7987 https://doi.org/10.1073/pnas.87.20.7983