한국산림과학회지 (Journal of Korean Society of Forest Science)

  • 제101권2호
  • /
  • Pages.291-296
  • /
  • 2012
  • /
  • 2586-6613(pISSN)
  • /
  • 2586-6621(eISSN)
한국산림과학회 (Korean Society of Forest Science)
권호 표지

유세포 분석을 통한 현사시나무 3배체 선발 및 계통별 기내생장 특성

Selection of a Triploid Poplar by Flow Cytometric Analysis and Growth Characteristics of its in vitro Grown Plants

  • 배은경 (국립산림과학원 산림생명공학과) ;
  • 이효신 (국립산림과학원 산림생명공학과) ;
  • 이재순 (국립산림과학원 산림생명공학과) ;
  • 노은운 (국립산림과학원 산림생명공학과)
  • Bae, Eun-Kyung (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Lee, Hyoshin (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Lee, Jae-Soon (Division of Forest Biotechnology, Korea Forest Research Institute) ;
  • Noh, Eun-Woon (Division of Forest Biotechnology, Korea Forest Research Institute)
  • 발행 : 2012.06.30
⟨ 이전 논문 다음 논문 ⟩

초록

세대기간이 긴 임목에 있어 3배체는 바이오매스 생산과 분자생물학적 연구 분야에서 매우 유용한 연구재료이다. 1970년대 육성된 현사시나무 4배체와 일반 2배체 간의 인공교배를 통해 3배체가 육성되었다. 유세포 분석법을 이용해 이들 $F_1$의 배수성을 확인한 결과 14개체 중 10개체가 3배체로 선발되었다. 그 가운데 3계통의 3배체(Line-1, Line-17, Line-18)는 기내배양시 2배체에 비해 잎이 크고, 엽형이 다소 변형된 형태적 특징이 있었다. 특히 'Line-18'은 줄기가 2배체에 비해 굵었으며, 뿌리 생육에 있어서 발근이 더뎠고 뿌리수도 다른 3배체나 2배체에 비해 적은 특징을 보였다. 그러나 순화율은 모두 100%에 달하였다. 본 연구결과는 3배체 현사시나무가 바이오매스 생산 및 형질전환 연구를 위한 좋은 재료가 될 수 있음을 보여주는 결과이며, 또한 유세포 분석법이 배수체 육종시 배수체를 확인하는데 효율적인 방법임을 시사한다.

Triploids are a useful tool for biomass production and molecular breeding of trees with a long life span. Triploids of the poplar 'Hyunsasi' (Populus alba ${\times}$ P. glandulosa) have been developed by crossing between female diploids and a male tetraploid. The tetraploid was developed around the 1970s at Korea Forest Research Institute by colchicine-induced chromosome doubling. Seedlings of the $F_1$ generation were analyzed using flow cytometry to verify their ploidy status. The mean relative fluorescence index of 3 F1 poplars, labeled as Line- 1, Line-17, Line-18, were approximately 1.5 times higher than those of diploid poplars, and the results clearly indicated that they were triploids. The phenotype of the F1 poplars included larger leaves and thicker stem than diploids, and abnormal leaf morphology, especially in the triploid 'Line-18'. Three triploid lines developed roots more slowly and had less roots than diploid. However, 3 poplar cytotypes (2x, Line-1, Line-17) rooted within 10 days on MS medium. In contrast, compared with the 3 cytotypes, the Line-18 showed about 80% and 70% in the rooting rate and the number of roots. The triploid poplars could be directly utilized for biomass production and with their sterility, they could serve as basic material for genetic transformation. In addition, flow cytometric analysis proved to be an effective and reliable method for screening forest trees for their ploidy level.

키워드

참고문헌

  1. Ahuja, M.R. and Neale, D.B. 2002. Origins of polyploidy in coast redwood [Sequoia sempervirens(D. Don) Endl.] and relationship of coast redwood to other genera of Taxodiaceae. Silvae Genetica 51(2-3): 93-100.
  2. Bae, E.K., Lee, H., Lee, J.S. and Noh, E.W. 2011. Drought, salt and wounding stress induce the expression of the plasma membrane intrinsic protein 1 gene in poplar. Gene 483(1-2): 43-48. https://doi.org/10.1016/j.gene.2011.05.015
  3. Bergstrom, I. 1940. On the progeny of diploid x triploid Populus tremula with special reference to the occurrence of tetraploidy. Hereditas 26: 191-201.
  4. Bradshaw, H.D. and Stettler, R.F. 1993. Molecular genetics of growth and development in Populus. I. Triploidy in hybrid poplars. Theoretical and Applied Genetics 86: 301-307.
  5. Chattopadhyay, S., Gandhi Doss, S., Halder, S., Ali, A.K. and Bajpai, A.K. 2011. Comparative micropropagation efficiency of diploid and triploid mulberry (Morus alba cv. S1) from axillary bud explants. African Journal of Biotechnology 10(79): 18153-18159.
  6. Eeckhaut, T., Leus, L. and Van Huylenbroeck, J. 2005. Exploitation of flow cytometry for plant breeding. Acta Physiolgiae Plantarum 27(4): 743-750. https://doi.org/10.1007/s11738-005-0079-2
  7. Ewald, D., Ulrich, K., Naujoks, G. and Schroder, M.B. 2009. Induction of tetraploid poplar and black locust plants using colchicine: chloroplast number as an early marker for selecting polyploids in vitro. Plant Cell, Tissue and Organ Culture 99(3): 353-357. https://doi.org/10.1007/s11240-009-9601-3
  8. Hu, W.J., Harding, S.A., Lung, J., Popko, J.L., Ralph, J., Stokke D.D., Tsai, C.J. and Chiang, V.L. 1999. Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nature Biotechnology 17(8): 808-811. https://doi.org/10.1038/11758
  9. Jia, C., Zhao, H., Wang, H., Xing, Z., Du, K., Song, Y. and Wei, J. 2004. Obtaining the transgenic poplars with low lignin content through down-regulation of 4CL. Chinese Science Bulletin 49(9): 905-909.
  10. Johnsson, H. 1945. The triploid progeny of the cross diploid x tetraploid Populus tremula. Hereditas 31: 411-440.
  11. Johnston, S.J., Bennett, M.D., Rayburn, L.A., Galbraith, D.W. and Price, J.H. 1999. Reference standards for determination of DNA content of plant nuclei. American Journal of Botany 86(5): 609-613. https://doi.org/10.2307/2656569
  12. Kang, X.Y. and Zhu, Z.T. 1997. A study on the 2n pollen vitality and germinant characteristics of white poplars. Acta Botanica Yunnanica 19(4): 402-406.
  13. Kang, X.Y., Zhu, Z.T. and Lin, H.B. 2000. Radiosensitivity of different ploidy pollen in poplar and its application. Acta Genetica Sinica 27(1): 78-82.
  14. Koo, Y.B. and Yeo, J.K. 2003. The status and prospect of poplar research in Korea. Journal of Korea Forest Energy 22(2): 1-17.
  15. Lee, H., Bae, E.K., Park, S.Y., Sjodin, A., Lee, J.S., Noh, E.W. and Jansson, S. 2007. Growth-phase-dependent gene expression profiling of poplar suspension cells. Physiologia Plantrum 131(4): 599-613. https://doi.org/10.1111/j.1399-3054.2007.00987.x
  16. Li Y.H., Kang X.Y., Wang S.D., Zhang Z.H. and Chen H.W. 2008. Triploid induction in Populus alba x P. glandulosa by chromosome doubling of female gametes. Silvae Genetica 57(1): 37-40.
  17. Muntzing, A. 1936. The chromosomes of a giant Populus tremula. Hereditas 21(2-3): 383-393.
  18. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassay with tabacco cultures. Physiologia Plantarum 15(3): 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  19. Niwa, Y. and Sasaki, Y. 2003. Plant self-defense mechanisms against oxidative injury and protection of the forest by planting trees of triploids and tetraploids. Ecotoxicology and Environmental Safety 55(1): 70-81. https://doi.org/10.1016/S0147-6513(02)00095-7
  20. Yang, S., Lu, L. and Ni, Y. 2006. Cloned poplar as a new fibre resource for the Chinese pulp and paper industry. Pulp & Paper Canada 107(2): 34-37.
  21. Zhang, Q., Zhang, Z.Y., Lin, S.Z. and Lin, Y.Z. 2005. Resistance of transgenic hybrid triploids in Populus tomentosa Carr. against 3 species of Lepidopterans following two winter dormancies conferred by high level expression of cowpea trypsin inhibitor gene. Silvae Genetica 54(3): 108-116.
  22. Zhang, Q., Zhang, Z.Y., Lin, S.Z., Zheng, H.Q., Lin, Y.Z., An, X.M., Li, Y. and Li, H.X. 2008. Characterization of resistance gene analogs with a nucleotide binding site isolated from a triploid white poplar. Plant Biology 10(3): 310-322. https://doi.org/10.1111/j.1438-8677.2008.00029.x
  23. Zhang, A.P., Liu, C.F. and Sun, R.C. 2010. Fractional isolation and characterization of lignin and hemicelluloses from triploid of Populus tomentosa Carr. Industrial Crops and Products 31(2): 357-362. https://doi.org/10.1016/j.indcrop.2009.12.003
  24. Zhiti, Z., Xiangyang, K. and Zhiyi, Z. 1998. Studies on selection of natural triploids Populus tomentosa. Sientia Slivae Sinicae 34(4): 22-31.