新疆珍稀植物阿魏内生菌与其共生机制及药效物质生成的相关性研究

Asafoetida was recorded in herbal book on materia medica since "The Tang Ben Cao". It was used by many national medicine, which had a long history and recorded in the calendar edition of "Pharmacopoeia of the People's Republic of China". Asafoetida was the endemic medicinal plant of Xinjiang, which have significant pharmacological effect and been included in the "The red book of plants in China--rare and endangered plants". At present, the artificial cultivation of Ferula has not realized. Modern research showed that endophytes were ubiquitous in plants, and there is interaction with the host. Applicants were isolated 337 strains of endophytic fungi, 178 strains of bacteria, 61 strains of Actinomycetes from Ferula in the completed project. We initially screened out the endophytic which had some promoting effect and significant pathogenic effect to aseptic seedling, and we also screened out the endophytic which could significantly promote the germination of Ferula seeds. We found that endophyte of Ferula had significant differences in different growth stages. Our study suggested that endophytes has some connection with the accumulation of medicinal material of Ferula. This study will examine the function and survival of endophyte, the mechanism of interaction with the Ferula, and the produced of medicinal material of Ferula contemplated by building symbiotic mode of endophytes and Ferula. Our research will provide a scientific basis for efficient use the medicinal substances of Ferula by biological technology. This has important implications for the sustainable utilization of Ferula, which was the rare and endangered resources in Xinjiang.

阿魏是自《唐本草》以来，历代本草均有记载的，应用历史悠久的，多民族医传统用药，并收载于历版《中华人民共和国药典》。现代药理作用显著，已列入中国珍稀濒危保护植物名录"的新疆特有药用植物资源。迄今为止尚未见实现阿魏人工栽培。现代研究表明植物普遍存在内生菌并与宿主存在着相互作用。申请者在已完成的基金项目中从阿魏植物中分离出内生真菌337株，细菌178株，放线菌61株；初步筛选出对阿魏无菌苗有一定促生作用和有明显致病性的菌群，及能够明显促进阿魏种子萌发的菌株；发现其在阿魏植物的不同生长期有明显的量的变化；提示内生菌与阿魏药效物质的积累存在某种关联。本项研究拟通过构建内生菌与阿魏共生培养模式；研究内生菌功能及生存方式，与作用阿魏植株的机理，与阿魏药效物质生成的相关性，为运用生物技术实现阿魏药效物质的高效利用提供科学依据。对发展新疆特有濒危珍稀资源阿魏的可持续利用具有重要的意义。

为了研究阿魏内生菌与其宿主之间的相互关系及在阿魏植株存在的部位及存在方式；探索阿魏内生菌与阿魏植株之间的作用机理，筛选促进阿魏生长的内生菌菌株；探索阿魏内生菌与阿魏药效物质生成的相关性，筛选促进阿魏药效物质生成的内生菌菌株。本研究在原有的内生菌的基础上，通过采集野生药用阿魏样品、提取分离内生菌、培养药用阿魏无菌苗、诱导愈伤组织及继代培养，完成了阿魏内生菌与阿魏无菌苗的共培养，筛选了90余株对阿魏生长有促进作用的内生菌，并重点对其中1株黑曲霉属的菌株代谢物促进小麦等作物种子萌发及幼苗生长的影响进行研究。本研究同时从阿魏植物中分离出内生菌337株，通过培养获得菌液，对其甲醇提取物进行抗肿瘤活筛选，并利用此模型将课题组分离获得的117份内生真菌提取物进行抗肿瘤活性筛选，已确定具有抗肿瘤活性的新疆阿魏内生真菌提取物49份，其中15份对Hela与AGS均具有显著抑制作用，且其抑制率达50%以上，重点选择抗肿瘤活性较强的菌株，对其进行代谢产物分离，共分离鉴定了10余个化合物，为从中内生菌中找到抗菌或抗肿瘤活性天然产物奠定基础。