Journal of agriculture & life science (농업생명과학연구)

Institute of Agriculture & Life Science, Gyeongsang National University (경상대학교 농업생명과학연구원)
권호 표지

Physical Wounding for the Enhancement of Agrobacterium-Mediated Transformation of Flammulina velutipes Mycelium

물리적 상해를 통한 Agrobacterium 이용 팽이균사체의 형질전환효율 증대

  • Duong, Van Thanh (Department of Biotechnology, Catholic University of Daegu) ;
  • Shin, Dong-Il (Department of Biotechnology, Catholic University of Daegu) ;
  • Park, Hee-Sung (Department of Biotechnology, Catholic University of Daegu)
  • 즈엉반탄 (대구가톨릭대학교 생명공학과) ;
  • 신동일 (대구가톨릭대학교 생명공학과) ;
  • 박희성 (대구가톨릭대학교 생명공학과)
  • Received : 2010.10.06
  • Accepted : 2010.12.17
  • Published : 2010.12.31
⟨ Previous Next ⟩

Abstract

In this study, Agrobacterium-mediated transformation was tested to the mycelium culture of Flamulina velutipes which is very popular as an edible mushroom in Korea. Particularly, aluminum oxide particles were used to generate wounds in F. velutipes mycelia via vigorous shaking prior to agro-infiltration. The result showed that transformants resistant to hygromycin could be obtained only from the mycelia with physical wounds. Gene transfer was verified by genomic DNA PCR. This study suggested a convenient tool to improve Agrobacterium-mediated transformation of F. velutipes.

본 연구에서는 국내에서 식용으로 재배되고 있는 팽이버섯의 균사체에 대하여 Agrobacterium을 이용한 형질전환을 시도하였다. 특히 물리적 연마제인 미세 aluminum oxide 입자를 팽이 균사체와 함께 강하게 교반함으로써 물리적 상해를 지니는 균사체를 제조하였으며 감압침윤에 의한 Agrobacterium 이용 형질전환을 시도하였다. Hygromycin 저항성을 이용한 선발 결과, 대조군에서는 균사체 생육이 전혀 관찰되지 않은 반면 물리적 상해군에서는 형질전환균사체 생육이 확인되었다. Genomic DNA PCR에 의한 유전자 도입을 확인함으로써 팽이균사체에 대한 매우 간편한 형질전환법을 제시할 수 있었다.

Keywords

References

  1. Bechtold, N. and G. Pelletier. 1998. In planta Agrobacterium-mediated gene transfer of adult Arabidopsis thaliana plants by vacuum infiltration. Meth. Mol. Biol. 82: 259-266.
  2. Cheng, M., J. E. Fry, S. Pang, I. Zhou, C. Hironaka, D. R. I. Duncan, T. W. L. Conner, and Y. Wang. 1997. Genetic transformation of wheat mediated by Agrobacterium tumefaciens, Plant Physiol. 115: 971-980.
  3. Cheng, M., T. Hu, J. L. Layton, C. N. Liu, and J. E. Fry. 2003. Desiccation of plant tissues post-Agrobacterium infection enhances T-DNA delivery and increases stable transformation efficiency in wheat. In Vitro Cell. Dev. Biol. Plant 39: 595-604. https://doi.org/10.1079/IVP2003471
  4. Chow, T. Y. K. and E. Kafer. 1993. A rapid method for isolation of total nucleic acids from Aspergillus nidulans. Fungal Genet. Newsl. 40: 25-27
  5. Enriquez-Obregon, G. A., R. I. Vazquez-Padron, D. L. Prieto-Samsonov, G. A. de la Riva, and G. Selman-Housein. 1998. Herbicide-resistant sugarcane (Saccharum officinarum L.) plants by Agrobacterium-mediated transformation. Planta 205: 20-27.
  6. Flores Solis, J. L., P. Mlejnek, K. Studena, and S. Prochazka. 2003. Application of sonication-assisted Agrobacterium-mediated transformation in Chenopodium rubrum. L. Plant Soil Environ. 49: 255-260.
  7. Kim, S., D. I. Shin, and H. S. Park. 2007. Transient $\beta$-glucuronidase expression in lily (Lilium longiflorum L.) pollen via wounding-assisted Agrobacterium- mediated transformation. Biotech. Lett. 29: 965-969. https://doi.org/10.1007/s10529-007-9326-5
  8. Kuo, C-Y., S-Y. Chou, R-S. Hseu, and C-T. Huang. 2010. Heterologous expression of EGFP in enoki mushroom Flammulina velutipes. Bot. Studies. 51: 303-309.
  9. Maehara T., M. Yoshida, Y. Ito, S. Tomita, K. Takabatake, H. Ichinose, and S. Kaneko. 2010. Development of a gene transfer system for the mycelia of Flammulina velutipes Fv-1 strain. Biosci. Biotechnol. Biochem. 74: 1126-1128. https://doi.org/10.1271/bbb.100021
  10. Meyer, V. 2008. Genetic engineering of filamentous fungi - progress, obstacles and future trends. Biotech. Adv. 26: 177-185. https://doi.org/10.1016/j.biotechadv.2007.12.001
  11. Olhoft, P. M, and D. A. Somers. 2001. L-cysteine increases Agrobacterium-mediated T-DNA delivery into soybean cotyledonary-node cells. Plant Cell Rep. 20: 706-711. https://doi.org/10.1007/s002990100379
  12. Pan, S. Q., T. Charles, S. Jin, Z. L. Wu, and E. W. Nester. 1993. Preformed dimeric state of the sensor protein VirA is involved in plant-Agrobacterium signal transduction Proc. Natl. Acad. Sci. USA 90: 9939-9943. https://doi.org/10.1073/pnas.90.21.9939
  13. Park, S-Y., A. F. van Peer, K-Y. Jang, P-G. Shin, Y. Park, Y-B. Yoo, K-M. Park, and W-S. Kong. 2010. Agrobacterium-mediated transformation using gill tissue of Flammulina velutipes. Kor. J. Mycol. 38: 48-53. https://doi.org/10.4489/KJM.2010.38.1.048
  14. Singh, N. and H. S. Chawla. 1999. Use of silicon carbide fibers for Agrobacterium-mediated transformation in wheat. Curr. Sci. 76: 1483-1485.
  15. Uze, M., J. Wunn, J. Pounti-Kaelas, I. Potrykus, and C. Sauter. 1997. Plasmolysis of precultured immature embryos improves Agrobacterium mediated gene transfer to rice (Oryza sativa L.) Plant Sci. 130: 87-95. https://doi.org/10.1016/S0168-9452(97)00211-2
  16. Wasser, S. P. and A. L. Weis. 1999. Medicinal properties of substances occurring in higher basidiomycetes mushrooms: Current perspectives (review). Int. J. Med. Mushr. 1: 31-62.