橡胶树树皮细胞木质素显示方法的比较分析  

Comparison of Different Staining Methods for Lignin in Bark of Rubber Tree(Hevea brasiliensis Muell.Arg.)

作　　者：史敏晶[1] 张世鑫[1] 丁欢 蒋毅 田维敏[1] SHI Minjing;ZHANG Shixin;DING Huan;JIANG Yi;TIAN Weimin(Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture and Rural Affairs/State Key Laboratory Incubation Base for Cultivation and Physiology of Tropical Crops,Haikou,Hainan 571101,China;Tibet Agricultural and Animal Husbandry University,Linzhi,Tibet 860000,China)

机构地区：[1]中国热带农业科学院橡胶研究所/农业农村部橡胶树生物学与遗传资源利用重点实验室/省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室,海南海口571101 [2]西藏农牧学院,西藏林芝860000

出　　处：《热带作物学报》2023年第10期2008-2015,共8页Chinese Journal of Tropical Crops

基　　金：海南省基金创新团队项目(No.320CXTD442);国家自然科学基金项目(No.31870590);现代农业产业技术体系建设专项(No.CARS-33-YZ1)。

摘　　要：巴西橡胶树是最重要的人工栽培产胶植物,树干割胶是当前从橡胶树乳管组织中获取天然橡胶的唯一途径。割胶后的排胶特性直接决定胶乳的产量,但其排胶机理目前尚不完全清楚。橡胶树树皮的膨压是乳管排胶的主要初动力,而细胞壁的主要成分木质素含量的高低与膨压密切相关。目前关于橡胶树树干树皮细胞中木质素含量的研究较少,缺乏通过形态结构研究分析木质素含量高低的研究。以橡胶树RY8-79和PR107成龄树树皮为研究材料,制备石蜡切片,分别以Wiesner反应方法、Maule反应方法以及番红染色结合荧光显色的方法对树皮组织中的木质素进行了染色定位分析。研究结果表明:光镜明场下,Wiesner反应方法和Maule反应方法能显色薄壁细胞壁中的木质素成分,但颜色浅淡,很难区分不同品系之间的差异;番红染色后能观察到薄壁细胞中浅紫红色的木质素成分,但不同组织以及不同品系之间同样很难区分;番红显色后结合546 nm绿光激发则能观察到木质素组织明显的红色荧光,不同细胞类型和不同品系之间差异明显,RY8-79薄壁细胞中木质素荧光明显弱于PR107,表明该品系中木质素的含量低于PR107。碘-溴染色显示次生乳管的结果表明,次生乳管分布的位置和木质素显色较深尤其是荧光强的部位一致。由此推测乳管及其周围的细胞中木质素含量较高,这一现象在PR107中尤其明显。此外,Wiesner反应和Maule反应不仅需要间苯三酚、盐酸和高锰酸钾(KMnO_(4))等危险化学试剂,而且反应时间不好把控,切片容易从载玻片上脱落以及细胞壁易破损;而番红染色结合荧光观察所用试剂则是安全无毒的且细胞结构完整。综合比较不同的方法,番红染色结合荧光显色是当前最适合橡胶树树皮组织细胞中木质素显色的方法,不仅能清晰区分木质素含量的高低,而且安全、简便易操作。Rubber tree is the most important cultivated rubber-producing plant,and the tapping of trunk is the only way to obtain natural rubber from the laticifer in the bark currently.The characteristics of latex flow after tapping determine the latex yield,but the mechanism of latex flow is not yet completely clear.The turgor pressure of bark is the main ini-tial driving force for latex flow,and the content of lignin,as the main component of cell wall,is closely related to the turgor pressure.At present,the study on the lignin content in bark of rubber tree is deficiency,among which the analysis of lignin content by morphological structure is lack.Using the bark of RY8-79 and PR107 mature tree as the research materials,paraffin sections were prepared,and the lignin in the bark was stained and localized by Wiesner reaction,Mäule reaction,and safranin staining combined with fluorescence,respectively.The results showed that both Wiesner reaction and Mäule reaction could stain the lignin component of the parenchyma cell wall,but difficult to distinguish different strains and tissues for the light color.The light purplish red of lignin in the parenchyma cell could be observed after safranin staining,but it was also difficult to distinguish different tissues and strains.The obvious red fluorescence of lignin tissue could be observed after safranin staining combined with 546 nm green light excitation,and there was significant difference among different cell types and strains.The lignin fluorescence of parenchyma cell in RY8-79 was significantly weaker than that in PR107,indicating that the content of lignin in RY8-79 was lower than that in PR107.Iodine-bromine staining showed that the distribution of secondary laticifer was consistent with the distribution of lignin with strong fluorescence,indicating that the lignin content of secondary laticifer and its surrounding parenchyma cells increased significantly,especially in PR107.In addition,Wiesner and Mäule reaction needed dangerous chemical re-agents such as phloroglucinol,h

关 键 词：巴西橡胶树 木质素 番红染色 薄壁细胞 荧光 

分 类 号：S794.1[农业科学—林木遗传育种]