荒漠植物牛心朴子耐盐内生真菌活性初筛  

Preliminary screening of salt tolerant endophytic fungi of desert plant Cynanchum hancockianum(Maxim.)Al.Iljinski

作　　者：段海婧 任远 马骏 陈靖[4,5] DUAN Haijing;REN Yuan;MA Jun;CHEN Jing(School of Pharmacy,Gansu University of Chinese Medicine,Lanzhou,Gansu,730101,China;Key Laboratory for Chinese Materia Medica Pharmacology and Toxicology in Gansu Province,Lanzhou,Gansu,730000,China;Pharmacology Laboratory of Chinese Materia Medica,Tertiary-level Laboratory for Traditional Chinese Medicine Scientific Research of National Administration of Traditional Chinese Medicine,Lanzhou,Gansu,730000,China;School of Pharmacy,Ningxia Medical University,Yinchuan,Ningxia,750004,China;Ningxia Hui Medicine Modernization Engineering Technology Research Center,Yinchuan,Ningxia,750004,China)

机构地区：[1]甘肃中医药大学药学院,甘肃兰州730101 [2]甘肃省中药药理与毒理学重点实验室,甘肃兰州730000 [3]国家中医药管理局中医药科研三级实验室中药药理实验室,甘肃兰州730000 [4]宁夏医科大学药学院,宁夏银川750004 [5]宁夏回药现代化工程技术研究中心,宁夏银川750004

出　　处：《甘肃中医药大学学报》2020年第4期23-29,共7页Journal of Gansu University of Chinese Medicine

基　　金：宁夏回族自治区科技攻关项目(2010168);教育部新世纪优秀人才支持计划(NCET-10-0915)。

摘　　要：目的考察含盐马铃薯葡萄糖琼脂(PDA)培养基对宁夏野生荒漠植物牛心朴子内生真菌分离、纯化及抗真菌活性的影响,为牛心朴子耐盐研究奠定基础。方法采用含不同质量分数NaCl(0%,5%,10%,15%,20%,30%)的PDA培养基对牛心朴子内生真菌进行分离、纯化;将得到的内生真菌发酵培养,发酵物分别用乙酸乙酯、正丁醇萃取后,采用稻瘟霉模型初步筛选萃取物活性菌株。结果共得到45株内生真菌,其中PDA培养基组15株(占全部菌株数的33.3%),5%NaCl组24株(占全部菌株数的53.3%),10%NaCl组6株(占全部菌株数的13.3%),NaCl质量分数大于15%时无菌株生长。乙酸乙酯部位完全抑制稻瘟霉孢子萌发共16株,其中PDA培养基组3株,且菌株Dy-3的活性极强(15.6μg/mL),几乎与酮康唑相当(7.8μg/mL),5%NaCl组共13株,10%NaCl组无活性;正丁醇部位完全抑制孢子萌发共18株,其中3株活性较强,且对应其乙酸乙酯部位活性较弱。结论耐盐培养对内生真菌的分离及抗稻瘟霉活性均有显著影响,5%NaCl为筛选耐盐活性菌株的较优质量分数;耐盐培养下正丁醇萃取物仍有活性,因此不能随意弃掉,以免漏筛。Objective To investigate saline potato dextrose agar(PDA)medium on isolation,purification of endophytic fungi from wild Cynanchum hancockianum(Maxim.)Al.Iljinski in Ningxia and its effects on antifungal activity,and to lay the foundation of study of salt tolerant endophytic fungi of desert plant Cynanchum hancockianum(Maxim.)Al.Iljinski.Methods PDA medium with different mass fraction of NaCl(0%,5%,10%,15%,20%,30%)was used to isolate and purify endophytic fungi from Cynanchum hancockianum(Maxim.)Al.Iljinski.The endophytic fungi obtained were fermented and cultured,and the fermentation products were extracted with ethyl acetate and n-butanol,respectively,and then the active strains of extracts were preliminarily screened by Magnaporthe grisea model.Results Totally 45 strains of endophytic fungi were obtained,including 15 strains in PDA medium group(accounting for 33.3%of the total number of strains),24 strains in 5%NaCl group(accounting for 53.3%of the total number of strains),and 6 strains in 10%NaCl group(accounting for 13.3%of the total number of strains).When the mass fraction of NaCl was greater than 15%,no strains grew.Totally 16 strains were completely inhibited spore germination of Magnaporthe grisea by ethyl acetate.Among them,3 strains were in PDA medium group,and strain Dy-3 had very strong activity(15.6μg/mL),almost equivalent to ketoconazole(7.8μg/mL),13 strains were in 5%NaCl group and 10%NaCl group had no activity.The n-butanol site completely inhibited the spore germination of 18 strains,among which 3 strains showed strong activity and their corresponding ethyl acetate site showed weak activity.Conclusion Salt-tolerant culture has significant effects on the isolation of endophytic fungi and antiactivity of Magnaporthe grisea,5%NaCl is the better mass fraction for screening salt tolerant strains;the n-butanol extract still has activity under salt tolerant culture,so it can not be discarded at will to avoid missing screening.

关 键 词：牛心朴子 荒漠植物 耐盐 内生真菌 稻瘟霉 

分 类 号：Q948.12[生物学—植物学]