JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2009, Vol. 33 ›› Issue (06): 44-.doi: 10.3969/j.jssn.1000-2006.2009.06.010

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Cryosection technique developed for stem tissue gene expression research on poplar and Chinese fir

LU Ye, XI Mengli, ZHENG Jia, SHI Jisen*   

  1. Key Laboratory of Forest Genetics and Biotechnology Ministry of Education, Nanjing Forestry University, Nanjing 210037, China
  • Online:2009-12-18 Published:2009-12-18

Abstract: Cryosection technique in woody plant becomes the key technology to investigate tissue differential expression and gene localization on woody plant genomic research. The cryosection protocols were developed on two model woody plants: Chinese fir and poplar. The stem samples were firstly processed using paraformaldehyde, acetone and Carnoy as fixative separately. Liquid Nitrogen can protect the tissue especially the tender one such as tissue culture’s materials. The tissue structure and cell morphology can be kept more properly by paraformaldehyde. By comparison, the cryosection was much better than other treatments, adapted with the 10% sucrose to the sample tissue, which has been fixed by acetone or Carnoy, and which can also be applied to the nucleic acid extraction. The adaptive treatment parameters of Chinese fir and poplar tissue culture seedling were 10% sucrose and then 50—90 seconds in liquid Nitrogen. The dehydration can make the cryosections remained clearer and intact. The tissues, which were more lignified, need to be softened by glycerol. The system established for Chinese fir and poplar stem can also be applied to other plant tissues.

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