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不同固定条件对几种植物样品超微结构的影响
引用本文:李叶,黄华平,邓睿,张新春,崔艳梅,卢雪莲,林培群.不同固定条件对几种植物样品超微结构的影响[J].热带作物学报,2016,37(11):2100-2105.
作者姓名:李叶  黄华平  邓睿  张新春  崔艳梅  卢雪莲  林培群
作者单位:1. 中国热带农业科学院环境与植物保护研究所,海南海口571101;农业部热带农林有害生物入侵监测与控制重点开放实验室,海南海口 571101;2. 中国热带农业科学院环境与植物保护研究所,海南海口,571101
基金项目:海南省重大专项(No. ZDZX2013023);中央级公益性科研院所基本科研业务费专项(No. 2015hzs1J017)。
摘    要:为比较几种固定条件对不同植物材料样品电镜超微结构图像清晰度的影响。以3个不同科属4种植物油棕(Elaeis gineansis Jacq.)、拟南芥(Arabidopsis thaliana.)、花生(Arachis hypogaea Linn.)、柱花草(Stylosanthes guianensias Sw.)]为研究主体代表,探讨了2.5%戊二醛与4%戊二醛固定过夜、锇酸固定2 h及过夜的制备方法对其叶片超微结构的影响,并成功观察了柱花草接种胶苞炭疽野生型菌株CH008后的病菌侵入过程。结果表明:油棕、拟南芥、花生及柱花草叶片经4%戊二醛与1%锇酸固定液固定过夜,其组织成像的效果均较为理想,各种细胞器整体信息丰富、结构组织完好、线条清晰。表明,此制备方法能够较好的保存植物叶片和细胞组织,提高制样的成功率及观察分辨率。

关 键 词:透射电镜  植物样品  制备方法  超薄切片  分辨率

The Improvement of the Sample Preparation Technique for TEM of Plant
LI Ye,HUANG Huaping,DENG Rui,ZHANG Xinchun,CUI Yanmei,LU Xuelian and LIN Peiqun.The Improvement of the Sample Preparation Technique for TEM of Plant[J].Chinese Journal of Tropical Crops,2016,37(11):2100-2105.
Authors:LI Ye  HUANG Huaping  DENG Rui  ZHANG Xinchun  CUI Yanmei  LU Xuelian and LIN Peiqun
Institution:1 Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, 2 Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture;1 Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, 2 Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture;Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences;1 Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, 2 Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture;1 Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, 2 Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture;Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences;1 Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, 2 Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture
Abstract:In this study, we analyzed the sample preparation from three different species for verifying available methods and application. For ultrastructural effect of Elaeis guineensis, Arabidopsis thaliana, Arachis hypogaea, Stylosanthes guianensias, we compared some methods, i.e. 2.5% of glutaraldehyde, 4% of glutaraldehyde, osmic acid(fixed for 2 h and overnight), and the infection process of wide -type strain CH008 in S. guianensias inoculated with Colletotrichum gloeosporioides was also observed. The results showed that there was no significant difference on the effect of ultramicro images of organelle when the tissues were fixed with 4% of glutaraldehyde and 2.5% of glutaraldehyde for A. thaliana, A. hypogaea and S. guianensias. However, for E. guineensis, the image effect of tissues treated with 4% of glutaraldehyde was better compared to the treatment of 2.5% of glutaraldehyde, and the effect of the leaves fixed for overnight was better than that for 2 h. Therefore, the leaf tissues would not get more injuries by the method(samples were fixed with 4% of glutaraldehyde and 1% osmic acid for overnight), which meaning that the method wouldl improve resolution ratio and be helpful for phytopathology studies.
Keywords:Transmission electron microscope  Sample preparation  Preparation method  Ultrathin section  Resolution
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