钌红染色和单克隆抗体免疫标记植物叶片中的果胶  

Ruthenium red staining and monoclonal antibody immunolabeling of pectin in plant leaves

作　　者：李晓宝 孙振炳 姚曜 汤正捷[1] 李晓平[1] LI Xiaobao;SUN Zhenbing;YAO Yao;TANG Zhengjie;LI Xiaoping(Yunnan Key Laboratory of Wood Adhesives and Glue Products,Southwest Forestry University,Kunming 650224,China)

机构地区：[1]西南林业大学云南省胶黏剂与胶合制品重点实验室,昆明650224

出　　处：《林业工程学报》2022年第3期100-106,共7页Journal of Forestry Engineering

基　　金：国家自然科学基金(31870551);云南省万人计划“青年拔尖人才”项目(YNWR-QNBJ-2018-120)。

摘　　要：果胶是植物组织中重要的化学组成成分之一,但目前关于植物叶片中果胶含量及表征的相关研究较少。为了更好地开发植物叶片的应用价值和发展潜力,利用草酸铵螯合法、钌红染色及单克隆抗体免疫标记法对6种植物叶片(杨树叶、云南松松针、刺槐叶、榆树叶、慈竹叶和大麻叶)中的果胶进行含量测定和表征。结果表明:大麻叶、杨树叶和榆树叶中的果胶含量(质量分数)较高,分别是12%~17%,10%~15%和10%~17%;刺槐叶和云南松松针次之,为5%~8%,1.5%~2.5%;竹叶最少,不足1%;确定了植物叶片中有大量的果胶存在。钌红染色和单克隆抗体免疫标记法均可有效表征果胶的分布,即植物叶片中的果胶部分可以被钌红染色,被单克隆抗体标记的果胶成分在紫外荧光照射下有亮光;比较着色程度和荧光亮度,可以看出果胶成分主要分布于叶片的叶肉细胞中。经过4种单克隆抗体标记之后,6种叶片中的荧光分布没有显著区别,即4种单克隆抗体能够标记的特定糖分在6种叶片中均有分布,果胶中的糖分是呈混合分布的状态,没有特定的富集区域。Pectin is one of the important chemical components in plant tissues,but there are limited related studies on the content and characterization of pectin in plant leaves.In order to increase the application value,the ammonium oxalate chelation method and ruthenium red staining and monoclonal antibody immunolabeling method were used to determine and characterize pectin in six plant leaves,i.e.,poplar(Populus L.) leaves,Pinus yunnanensis leaves,Robinia(Robinia pseudoacacia L.) leaves,elm(Ulmus pumila L.) leaves,bamboo(Neosinocalamus affinis) leaves and hemp(Cannabis sativa L.) leaves.The results showed that the hemp leaves,poplar leaves and elm leaves had high pectin contents of 12%-17%,10%-15% and 10%-17%,respectively.Robinia leaves and Pinus yunnanensis leaves followed by 5%-8% and 1.5%-2.5%,and bamboo leaves had the smallest content,which was less than 1%.It was confirmed that there was a large amount of pectin in plant leaves.Both ruthenium red staining and monoclonal antibody immunolabeling methods can effectively characterize the distribution of pectin,that is,the pectin part of plant leaves can be stained with ruthenium red,and the pectin component labeled with monoclonal antibody would have bright light under the ultraviolet fluorescence irradiation.The color depth and fluorescence brightness were directly proportional to the amount of pectin content.Comparing the coloring and fluorescence conditions of the six leaves,hemp leaves,Robinia leaves,poplar leaves,and elm leaves had more pectin content than the other two,followed by Pinus yunnanensis.Bamboo leaves were the least pectin content among the 6 types of leaves,which is consistent with the results of the determination of pectin content by ammonium oxalate chelation.Therefore,hemp leaves,poplar leaves and elm leaves had the potential to extract pectin.The four kinds of monoclonal antibody-labeled hemp leaves,Robinia leaves,elm leaves and poplar leaves mesophyll cells had brighter fluorescence and color depth than epidermal cells,that is,the mesophyll cells

关 键 词：果胶 植物叶片 钌红染色 单克隆抗体免疫标记 

分 类 号：S718.47[农业科学—林学]