丹参潜在性致病内生真菌的筛选及其致病相关酶酶学特性分析  

Screening of Potentially Pathogenic Endophytic Fungi of Salvia miltiorrhiza and Analysis of Their Relevant Enzymatic Characteristics

作　　者：林亚丽 黄倩倩 严铸云[1] 陈新[1] 王海[1] 何冬梅[1] 韩桂琪[1] 

机构地区：[1]成都中医药大学中药材标准化省部共建教育部重点实验室四川省中药资源系统研究与开发利用省部共建国家重点实验室

出　　处：《中国实验方剂学杂志》2018年第4期38-43,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81573537)

摘　　要：目的：从18株丹参内生真菌中筛选具有潜在性致病作用的内生真菌,并对其致病机制进行探究。方法：采用内生真菌与组培苗共培养方式,根据组培苗的发病情况筛选出潜在性致病作用的内生真菌,测定活体外和活体根内的纤维素酶[羧甲基纤维素酶（Cx）,β-葡萄糖苷酶（β-G）],果胶酶[多聚半乳糖醛酸酶（PG）,聚甲基半乳糖醛酸酶（PMG）]和半纤维素酶（木聚糖酶）等细胞壁降解酶（CWDE）活性的变化。结果：在活体外,PG,PMG,木聚糖酶活性先增加后降低到后期基本不变,Cx,β-G酶活性随着培养时间的增加而持续升高。在活体内,9株潜在性致病内生真菌均能在组培苗根部组织产生细胞壁降解酶,但不同菌种差异较大。其中7株菌接种的组培苗根部Cx,β-G活性显著高于未接菌的根部（P〈0.05）,6株菌接种的组培苗根部PG活性显著高于未接菌的根部（P〈0.05）,9株菌接种的组培苗根部PMG以及木聚糖酶活性显著高于未接菌的根部（P〈0.05）。结论：Cx,β-G和PG,PMG以及木聚糖酶在潜在性致病内生菌的致病过程中起到了重要作用,是重要的致病因子,但各潜在性致病内生真菌的作用环节存在差异。Objective： To screen out the potentially pathogenic endophytic fungi from 18 strains of endophytic fungi isolated from roots of Salvia miltiorrhiza,in order to explore the pathogenic mechanism of these fungi.Method： The plant-endophyte symbotic experiment was conducted to choose the potential pathogens.To determine pathogenic fungi,the changes in the activities of cell wall degrading enzymes,including cellulose[carboxymethyl cellulose enzyme（Cx）,β-glucosidase（β-G） ],pectinase [polygalacturonase（PG）,polymethylgalacturonase（PMG） ] and xylanase enzymes were measured in vitro and in vivo.Result： In vitro,all of the 9 strains could secrete cell wall degrading enzymes（CWDE）,including cellulose,pectinase and xylanase.Cx and β-G activities increased over time,while PG,PMG and xylanase activities first increased and then decreased.In vivo,all of the 9 strains could secrete cell wall degrading enzymes in the roots of Salvia miltiorrhiza,but with differences among them.Roots of inoculations of 7 strains showed increase in Cx and β-G activities,roots of inoculations of 6 strains showed increase in PG activity,while roots of inoculations of 9 strains showed higher PMG and xylanase activities.Conclusion： Cellulase and pectinase and xylanase play important roles as pathogenic factors,but with differences in actions of potentially pathogenic endophytic fungi.

关 键 词：丹参 潜在性致病内生真菌 细胞壁降解酶 

分 类 号：R282[医药卫生—中药学] Q939.5[医药卫生—中医学]