Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Quantitative Analysis of Transient Expression in Tah Tasai Chinese Cabbage (Brassica campestris var. narinosa) Seedlings Following Agrobacterium-Mediated Transformation

다채 (Brassica campestris var. narinosa) 유묘의 형질전환 및 일시발현의 정량적 분석

  • Shin Dong-Il (Department of Biotechnology, Catholic University of Daegu) ;
  • Park Hee-Sung (Department of Biotechnology, Catholic University of Daegu)
  • 신동일 (대구가톨릭대학교 생명공학과) ;
  • 박희성 (대구가톨릭대학교 생명공학과)
  • Published : 2005.12.01
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Abstract

Tah tasai chinese cabbage (Brassica campestris var. narinosa), a vegetable plant popularly consumed as several-days-old seedlings in oriental countries, can be easily cultivated using a simple appliance. We demonstrated that Agrobacterium-mediated transformation via vacuum infiltration (agroinfiltration) resulted in a successful transient GUS gene expression in tah tasai chinese cabbage seedlings. Pre-germinated seeds were found to be more susceptible to Agrobacterium infection than one-day-old or two-days-old seedlings. We also demonstrated that hydrogen peroxide (HPO) treatment increased GUS expression especially for two-days-old seedlings. In ELISA using seedlings transformed with hepatitis B surface antigen (HBsAg) DNA by agroinfiltration, HBsAg protein synthesis increased more than two folds by HPO treatment to two-days-old seedlings in comparison to the mock-treated pre-germinated seeds.

Brassica campestris var. narinosa는 새싹채소로 애용되는 식물로서 간단한 용기에서 손쉽게 재배할 수 있다. 본 연구에서는 vacuum-infiltration을 통한 Agrobacterium-mediated transformation에 의해서 B. campestris var. narinosa 싹에서의 GUS일시발현을 성공적으로 확인하였다. 발아전의 종자가 발아를 이미 시작한 종자보다 그 형질전환 효율이 높게 나타났다. 한편, hydrogen peroxide를 2일 생장시킨 싹에 처리한 후 형질전환 하였을 때 GUS발현이 증가되는 것을 관찰하였다. 간염항원유전자의 형질전환과 ELISA에 의한 항원단백질의 발현량 분석 시 hydrogen peroxide 처리 싹이 비처리 형질전환 싹에서 보다 2배 이상의 발현량이 측정되었다.

Keywords

References

  1. Bechtold N. Ellis J, Pelletier, G (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidapsis thaliana plants. CR Acad Sci Paris 316: 1194
  2. De Block M, De Brower D, Tenning P (1989) Transformation of Brassica napus and Brassica oleacea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants. Plant Physiol 91: 694-701 https://doi.org/10.1104/pp.91.2.694
  3. Doran PM (2000) Foreign protein production in plant tissue cultures. Curr Opin Biotech 11: 199-204 https://doi.org/10.1016/S0958-1669(00)00086-0
  4. Fisher R, Vaquero-Martin C, Sack M, Drossard J, Emans N, Commandeur U (1999) Towards molecular farming in the future: transient protein expression in plants. Biotechnol Appl Biochem 30: 113-116
  5. Giddings G (2001) Transgenic plants as protein factories. Curr Opin Biotech 12: 450-454 https://doi.org/10.1016/S0958-1669(00)00244-5
  6. Jain RK, Chowdhury JK, Charma DR, Friedt W (1988) Genotypic and media effects on plant regeneration from cotyledon explant cultures of some Brassica species. Plant Cell Tissue Organ Cult 14: 197-206 https://doi.org/10.1007/BF00043410
  7. Jefferson RA (1987) Assaying chimeric genes in plants: the gus gene fusion system. Plant Mol Biol Rep 5: 387-405 https://doi.org/10.1007/BF02667740
  8. Jun SI, Kwon SY, Paek KY, Paek KH (1995) Agrobacterium-mediated transformation and regeneration of fertile transgenic plants of chinese cabbage (Brassica campestris ssp. pekinensis cv ‘Spring flavor’). Plant Cell Rep 14: 620-625 https://doi.org/10.1007/BF00232725
  9. Knutzon DS, Thompson GA, Radke SE, Jhonson WB, Knauf VC, Kridl JC (1992) Modification of Brassica seed oil by antisense expression of a stearoyl-acyl carries protein desaturase gene. Proc Natl Acad Sci USA 89: 2624-2628 https://doi.org/10.1073/pnas.89.7.2624
  10. Mukhopadhyay A, Armugam N, Nandakumar PBA, Pradhan AK, Gupta V, Pental D (1992) Agrobacterium-mediated transformation of oil seed Brassica campestris: transformation frequency is strongly influenced by the mode of shoot regeneration. Plant Cell Rep 11: 506-513
  11. Narashimhulu SB, Chopra VL (1988) Species specific shoot regeneration response of Brassicas. Plant Cell Rep 7: 104-106 https://doi.org/10.1007/BF00270115
  12. Radke SE, Andrews BM, Moloney MM, Crouch ML, Kridl JC, Knauf VC (1988) Transformation of Brassica napus L. using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene. Theor Appl Genet 75: 685-694
  13. Swanson EB, Erickson LR (1989) Haploid transformation in Brassica napus using an octopine-producing strain of Agrobacterium tumefaciens. Theor Appl Genet 78: 831-835
  14. Vitha S, Benes K, Phillips JP, Gartland KMA (1995) Histochemical GUS analysis. In Agrobacterium Protocols, K. M. A. Gartland and M. R. Davey, eds (Totowa, NJ: Humana Press), pp 185-193
  15. Xiang Y, Wong WKR, Ma MC, Wong RSC (2000) Agrobacterium-mediated transformation of Brassica campestris ssp. Parachinensis with synthetic Bacillus thuringiensis cry1Ab and cry1Ac genes. Plant Cell Rep 19: 251-256 https://doi.org/10.1007/s002990050007