[1]陆叶,席梦利,郑佳,等.杨树和杉木茎段组织的冰冻切片技术研究[J].南京林业大学学报(自然科学版),2009,33(06):044.[doi:10.3969/j.jssn.1000-2006.2009.06.010]
 LU Ye,XI Meng li,ZHENG Jia,et al.Cryosection technique developed for stem tissue gene expression research on poplar and Chinese fir[J].Journal of Nanjing Forestry University(Natural Science Edition),2009,33(06):044.[doi:10.3969/j.jssn.1000-2006.2009.06.010]
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杨树和杉木茎段组织的冰冻切片技术研究
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《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
33
期数:
2009年06期
页码:
044
栏目:
研究论文
出版日期:
2009-11-30

文章信息/Info

Title:
Cryosection technique developed for stem tissue gene expression research on poplar and Chinese fir
作者:
陆叶席梦利郑佳施季森*
南京林业大学,林木遗传与生物技术省部共建教育部重点实验室,江苏南京210037
Author(s):
LU Ye XI Mengli ZHENG Jia SHI Jisen*
Key Laboratory of Forest Genetics and Biotechnology Ministry of Education, Nanjing Forestry University, Nanjing 210037, China
关键词:
杨树杉木冰冻切片基因表达
Keywords:
Populus Cunninghamia lanceolata(Lamb.)Hook cryosection
分类号:
S722
DOI:
10.3969/j.jssn.1000-2006.2009.06.010
文献标志码:
A
摘要:
以木本植物中的模式树种杨树和杉木茎段为试验材料,分别用多聚甲醛、丙酮和卡诺固定液,研究了这两个树种的冰冻切片技术体系。结果表明,液氮冷冻法对幼嫩的组织(如组培苗)有较好的保护作用。多聚甲醛固定液可以较好地保存细胞的形态结构,冰冻切片较完整。丙酮和卡诺固定液加10%蔗糖保护之后效果更好,切片完整且可以进一步用于核酸提取。卡诺氏固定液固定后,杉木和杨树组培苗的蔗糖保护剂适宜质量分数为10%,液氮冷冻时间为50~90 s。脱水步骤可以使冰冻切片更清晰完整。而田间植株木质化程度较高的茎段组织则需要甘油软化辅助处理,即可获得清晰的冰冻切片。
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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备注/Memo

备注/Memo:
收稿日期:2009-03-27修回日期:2009-08-03基金项目:江苏省自然科学基金资助项目(BK2007216);江苏省科技攀登计划(BK2008051);南京林业大学科技创新基金项目(cx070101)作者简介:陆叶(1982—),助理实验师。*施季森(通讯作者),教授,研究方向为林木遗传育种,林木分子生物学。Email: jshi@njfu.edu.cn。引文格式:陆叶,席梦利,郑佳,等. 杨树和杉木茎段组织的冰冻切片技术研究[J]. 南京林业大学学报:自然科学版,2009,33(6):44-48.
更新日期/Last Update: 2009-11-30