朱波 刘京晶 斯金平 秦路平 韩婷 赵鹂 吴令上

[摘要]为揭示初筛所得7株活树附生铁皮石斛内生真菌对宿主生长及代谢成分的影响，该研究观察测量了接菌与未接菌铁皮石斛组培苗形态特征与农艺性状，采用苯酚硫酸法与热浸法测定组培苗茎总多糖含量与浸出物含量，柱前衍生HPLC分析茎多糖中单糖组成，HPLC分析茎醇溶性成分。研究表明，供试7个菌株可改变铁皮石斛组培苗茎节变紫进程，并影响茎糖类及醇溶性成分组成及含量积累，除DO34外均能促进组培苗生长与提高多糖含量，除DO12外均能促进甘露糖含量积累，DO18，DO19，DO120可促进醇溶性浸出物含量积累，综合生长及代谢成分多项指标筛选出DO14，DO18，DO19与DO120 4个优良菌株，为铁皮石斛内生真菌和宿主相互作用机制研究与微生物工程研究提供了菌种材料与研究基础。

[关键词]铁皮石斛； 内生真菌； 生长发育； 代谢成分

[Abstract]The paper aims to study the effects of endophytic fungi from D officinale cultivated on living trees on growth and components metabolism of tissue culture seedlings Morphological characteristics and agronomic characters of tissue culture seedlings infected and uninfected by endophytic fungus were observed and measured Polysaccharides and alcoholsoluble extracts contents were determined by phenolsulfuric acid method and hotdipmethod， respectively Monosacchride composition of polysaccharides and alcoholsoluble extracts components were analyzed by precolumn derivatives HPLC and HPLC method， respectively It showed that effects of turning to purple of stem nodes could be changed by endophytic fungus Besides， the endophytic fungus could affect the contents and constitutions of polysaccharides and alcoholsoluble extracts The strains tested， expect DO34， could promote growth and polysaccharides content of tissue culture seedlings The strains tested， expect DO12， could promote the accumulation of mannose Furthermore， DO18， DO19 and DO120 could increase alcoholsoluble extracts On the basis， four superior strains were selected for mechanism research between endophytic fungus and their hosts and microbiology engineering

[Key words]Dendrobium officinale； endophytic fungi； growth and development； components metabolism

doi：10.4268/cjcmm20160906

中藥材的品质与其生长环境密切相关，生长环境包括生物性因素与非生物性因素，植物内生真菌作为一个重要的生物性因素已经越来越受到国内外学者的关注[14]，现有研究表明植物内生真菌能促进宿主生长、提高宿主对生物胁迫或非生物胁迫的抗性[5]，能参与宿主次生代谢产物的合成和积累，影响药用植物的品质[67]。

铁皮石斛Dendrobium officinale Kimura et Migo是我国传统名贵中药材，具有益胃生津、滋阴清热等功效，用于热病津伤，口干烦渴，胃阴不足，食少干呕等症，收载于2015年版《中国药典》[8]。近年来，铁皮石斛内生真菌研……