宁夏黄花棘豆中产苦马豆素内生真菌的分离及鉴定  

Isolation and Identification of Swainsonine-producing Fungal Endophyte from Oxytropis ochrocephala Bunge in Ningxia

作　　者：张蕾蕾[1] 何生虎[1] 余永涛[1] 赵清梅[2,3] 

机构地区：[1]宁夏大学农学院,宁夏银川750021 [2]北方民族大学生物科学与工程学院,宁夏银川750021 [3]国家民委发酵酿造工程生物技术重点实验室,宁夏银川750021

出　　处：《中国畜牧兽医》2013年第12期165-172,共8页China Animal Husbandry & Veterinary Medicine

基　　金：宁夏自然科学基金(NZ1118);国家自然科学基金(31201962)

摘　　要：对宁夏黄花棘豆中产苦马豆素内生真菌进行分离和鉴定,为进一步研究疯草内生真菌合成苦马豆素的分子调控机理提供菌种和奠定基础。分离并纯化黄花棘豆植物组织中的真菌,采用气相色谱质谱联用法对各菌株菌丝中的苦马豆素进行分析,筛选能产生苦马豆素的内生真菌;紫外线照射和黑暗交替培养诱导内生真菌产孢;提取真菌DNA,扩增和测定真菌甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GPD)、内部转录间隔区(internal transcribed space,ITS)和线粒体核糖体小亚基(mitochondrial small subunit,mt SSU)基因序列,进行同源性比对和遗传进化分析,对其进行种属分类和命名。本研究从宁夏黄花棘豆中共分离到5株真菌,其中菌株NX-FEL001菌丝中含有苦马豆素,经诱导培养,该菌株能产生分生孢子及分生孢子梗等结构;序列同源性和系统发育分析结果表明,该菌株与Undifilum真菌的亲缘关系最近,根据形态结构和分子进化分析结果将菌株NX-FEL001鉴定为Undifilum oxytropis。To provide strains and a novel knowledge for the further study of the molecular regulation mechanism of endophytic fungi produced swainsonine in locoweed, this research was performed to isolate and identify swainsonine-producing fungal endophytes from Oxytropis ochrocephala Bunge in Ningxia. The fungal endophytes from the plant tissues were isolated and purified. Each strain was analyzed with gas chromatography-mass spectrum to determine the presence of swainsonine in fungal mycelia.The swainsonine-producing strain was induced to produce conidia or conidiospores in the culture condition of ultraviolet light and darkness alternation. The fungal DNA were extracted from mycelia of the swainsonine-producing fungal endophyte. The GPD, ITS and mt SSU of the strain were amplified and sequenced for the analysis of homology and genetic evolution. The taxon of the strain was determined based on the morphology and the result of genetic evolution analysis. The results indicated that five isolates were harvested from Oxytropis ochrocephala Bunge in Ningxia. Only one strain was determined to synthesize swainsonine. The strain could produce abundant conidia or conidiospores after induced culture. The morphology and genetic evolution relationship of the strain were closely related to the Undifilum. Therefore, the strain was identified as Undifilum oxytropis in this study.

关 键 词：黄花棘豆 内生真菌 苦马豆素 Undifilure OXYTROPIS 

分 类 号：S816.79[农业科学—饲料科学]