| 摘要: |
| 【目的】探索瘤菌根菌与铁皮石斛共生后对β-1,4-甘露糖苷酶基因DoMan1表达和甘露糖含量的影响，为铁皮石斛高品质栽培和药用成分的开发研究提供思路。【方法】从前期构建的瘤菌根菌与铁皮石斛共生的转录组数据库中获得差异表达基因DoMan1的序列，通过理化性质、结构特点和亚细胞定位进行蛋白功能分析，并通过qPCR和HPLC分别检测瘤菌根菌与铁皮石斛共生后DoMan1的时空表达模式和甘露糖含量的变化。【结果】DoMan1的ORF为1 326 bp，可编码441个氨基酸，属于β-1,4-甘露糖苷酶家族成员;DoMan1蛋白与金钗石斛的同源蛋白具有最近的进化关系，主要定位于细胞膜和内质网。旺盛期和封顶期是铁皮石斛多糖积累的重要时期，甘露糖以多糖形式储存在茎、叶中。DoMan1在铁皮石斛根、茎、叶组织中均有表达，其中在叶片和茎中的表达量较高。当瘤菌根菌与根共生后，DoMan1的表达量比对照显著提高，游离甘露糖和总甘露糖含量也显著增加，且封顶期茎、叶中游离甘露糖含量较高。【结论】瘤菌根菌与铁皮石斛共生可诱导DoMan1表达，显著提高铁皮石斛茎、叶中的甘露糖含量。 |
| 关键词: 铁皮石斛 瘤菌根菌 甘露糖 甘露糖苷酶 |
| DOI：10.13207/j.cnki.jnwafu.2025.11.010 |
| 分类号: |
| 基金项目:福建省公益类竞争性项目(2023R1093)；福建省自然科学基金项目(2024J01341) |
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| Effects of Epulorhiza sp. on the expression of β-1,4-mannosidase gene DoMan1 and mannose content in Dendrobium officinale |
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WANG Weiying, ZOU Hui, LIN Jiangbo, DAI Yimin
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Subtropical Agriculture Research Institute,Fujian Academy of Agricultural Sciences,Zhangzhou,Fujian 363005,China
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| Abstract: |
| 【Objective】Expression of the β-1,4-mannosidase gene (DoMan1) and mannose accumulation were investigated when a symbiotic relationship was established between Dendrobium officinale with Epulorhiza sp..This study aimed to provide insights into the high-quality cultivation of D.officinale and the development of medicinal ingredients.【Method】Sequence of the differentially expressed gene DoMan1 was obtained from previous transcriptome data of the symbiosis between Epulorhiza sp.and D.officinale. Protein function was analyzed through analyzing its physical and chemical properties,structural characteristics,and subcellular localization. Additionally,DoMan1 expression was detected using qPCR and mannose content was determined by HPLC.【Result】The ORF of DoMan1 was 1 326 bp,encoding 441 amino acids,which belongs to the β-1,4-mannosidase family.DoMan1 protein had the closest evolutionary relationship with the homologous protein of D.nobile.DoMan1 was located at cell membrane and endoplasmic reticulum.The vigorous growth and apical-closure periods were important periods of polysaccharide accumulation in D.officinale,and mannose was stored in the form of polysaccharides in the stems and leaves.DoMan1 was expressed in all tissues,with the highest expression level in the leaves.After the symbiosis between Epulorhiza sp.and D.officinale roots,the expression of DoMan1 was induced,indicating that this gene was induced by Epulorhiza sp.infection.The polysaccharides in D.officinale were mainly composed of mannose and glucose,and Epulorhiza sp.infection promoted mannose accumulation,with the highest content in stems.Free mannose in stems and leaves during the apical-closure period was relatively high.【Conclusion】DoMan1 expression was induced and mannose content in stems and leaves was increased after the establishment of symbiosis between Epulorhiza sp.and D. officinale. |
| Key words: Dendrobium officinale Epulorhiza sp. mannose mannosidase |
