Journal of agriculture & life science (농업생명과학연구)

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

Effect of Cell Wall-Wounding Reagents on Agrobacterium-mediated Barley Seedling Transformation

Agrobacterium 이용 보리묘 형질전환에 대한 세포벽 상해물질의 효과

  • 최장원 (대구대학교 바이오산업전공) ;
  • 박희성 (대구가톨릭대학교 생명공학과)
  • Received : 2010.01.13
  • Accepted : 2010.02.22
  • Published : 2010.02.28
⟨ Previous Next ⟩

Abstract

Barley, a monocotyledonous plant, is relatively recalcitrant to the process of Agrobacterium-mediated genetic transformation. In this study, seedlings of six barley cultivars (Keunal-1-Ho, Saessal, Ol, Saechalssal, Seodunchal and Pungsanchalssal) were injured using alkali, oxidizing or reducing agents. They were then transformed using Agrobacterium via vacuum infiltration for the analysis of comparative GUS gene expression. It was determined that chemical injuries causing a slight growth retardation could overall enhance the GUS transformation rate. Hydrogen peroxide was determined to be the most effective.

단자엽 식물인 보리는 Agrobacterium을 이용한 형질전환이 비교적 까다로운 편이다. 본 연구에서는 큰알1호, 내쌀보리, 올보리, 새찰쌀보리, 서둔찰보리, 풍산찰쌀보리의 유묘에 알칼리, 산화제, 환원제 등을 처리하여 화학적 상처를 유발하였으며 이들에 감압진공을 이용한 Agrobacterium 형질전환을 실시한 후 GUS 유전자발현을 분석하였다. 그 결과, 보리묘 생육을 일부 저하시킬 수 있는 농도의 화학물질 처리는 각기 다른 보리 품종의 형질전환율을 전반적으로 증대시킬 수 있는 것으로 판단되었다. 화학물 중에서는 특히 hydrogen peroxide 처리가 비교적 우수한 것으로 나타났다.

Keywords

References

  1. Ahlandsberg, S., P. Sathish, C. X. Sun, and C. Jansson. 1999. Green fluorescent protein as a reporter system in the transformation of barley cultivars. Physiol. Plant 107: 194-200. https://doi.org/10.1034/j.1399-3054.1999.100207.x
  2. Arencibia, A. D., E. R. C. Carmona, P. Tellez, M. T. Chan, S. M. Yu, L. E. Trujillo, and P. Oramas. 1998. An efficient protocol for sugarcane ( Saccharum spp. L) transformation mediated by Agrobacterium tumefaciens. Transgenic Res. 7: 213-222. https://doi.org/10.1023/A:1008845114531
  3. Bartlett, J. G., S. C. Alves, M. Smedley, J. W. Snape, and W. A. Harwood. 2002. Highthroughput Agrobacterium-mediated barley transformation. J. Plant Physiol. 159: 1131- 1138. https://doi.org/10.1078/0176-1617-00707
  4. 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.
  5. 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. https://doi.org/10.1104/pp.115.3.971
  6. 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: 595-604l. https://doi.org/10.1079/IVP2003471
  7. 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.
  8. Flores Solis, J. L., P. Mlejnek, K. Studena, and S. Proch zka. 2003. Application of sonication- a assisted Agrobacterium-mediated transformation in Chenopodium rubrum. L. Plant Soil Environ. 49: 255-260. https://doi.org/10.1080/00380768.2003.10410005
  9. Floros, J. D., H. Y. Wetzstein, and M. S. Chinnan. 1987. Chemical (NaOH) peeling as viewed by scanning electron microscopy: pimiento peppers as a case study. J. Food Sci. 52: 1312-1316. https://doi.org/10.1111/j.1365-2621.1987.tb14071.x
  10. Harwood, W. A., S. M. Ross, P. Cilento. and J. W. Snape. 2000. The effect of DNA/gold particle preparation technique, and particle bombardment device, on the transformation of barley ( Hordeum vulgare). Euphytica 111: 67-76. https://doi.org/10.1023/A:1003700300235
  11. Hiei, Y., S. Ohta, T. Komari, and T. Kumashiro. 1994. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6: 271-282 https://doi.org/10.1046/j.1365-313X.1994.6020271.x
  12. Ishida, Y., H. Saito, S. Ohta, Y. Hiei, T. Komari, and T. Kumashiro. 1996. High efficiency transformation of maize ( Zea mays L.) mediated by Agrobacterium tumefaciens. Nature Biotechnol. 14: 745-750. https://doi.org/10.1038/nbt0696-745
  13. Jefferson, R. A. 1987. Assaying chimeric genes in plants: the gus gene fusion system. Plant Mol. Biol. Rep. 5: 387-405. https://doi.org/10.1007/BF02667740
  14. 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
  15. May, G. D., R. Afza, H. S. Mason, A. Wiecko, F. J. Novak, and C. J. Arntzen. 1995. Generation of transgenic banana ( Musa acuminata) plants via Agrobacterium-mediated transformation. Bio/Technology. 13: 486-492. https://doi.org/10.1038/nbt0595-486
  16. 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
  17. 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
  18. Riva, G. A., J. Gonzalez-Cabrera, J. Vasqu- Padru, and C. Ayra-Pardo. 1998. Agrobacterium tumefaciens gene transfer to plant cell. Elect. J. Biotechnol. 2: 488-495.
  19. Shin, D. I., and H. S. Park. 2005. Transient expression in Chinese cabbage by hydrogen peroxide-aided agroinfiltration. Agric. Chem. Biotech. 48: 229-230.
  20. Singh, N., and H. S. Chawla. 1999. Use of silicon carbide fibers for Agrobacterium-mediated transformation in wheat. Curr. Sci. 76: 1483-1485.
  21. Tingay, S., D. McElroy, R. Kalla, S. Fieg, M. Wang, and R. Brettel. 1997. Agrobacterium-mediated barley transformation. Plant J. 11: 1369-1376. https://doi.org/10.1046/j.1365-313X.1997.11061369.x
  22. 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
  23. Watts, M. E., I. J. Roberts, and M. A. Woodcock. 1989. Comparison of colorimetric and clonogenic assays for hypoxic-specific toxins with hamster and human cells. Int. J. Radiat. Oncol. Bio. Phys. 16: 939-942. https://doi.org/10.1016/0360-3016(89)90891-2