Please wait a minute...
浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2015, Vol. 41 Issue (5): 586-592    DOI: 10.3785/j.issn.1008-9209.2015.03.031
资源与环境科学     
石油污染土壤中产苦参碱真菌的分离与鉴定
毛东霞, 郭丹丹, 吴玲玲, 田晓雪, 张胜翔, 刘陶, 马小魁
1.陕西师范大学生命科学学院/西北濒危药材资源开发国家工程实验室,西安710100;2.河南省疾病预防控制中心,郑州450000
Isolation and identification of a matrine-producing fungal strain from petroleum-contaminated soil
Mao Dongxia, Guo Dandan, Wu Lingling, Tian Xiaoxue, Zhang Shengxiang, Liu Tao, Ma Xiaokui
(1.College of Life Sciences, Shaanxi Normal University/National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China, Xi’an 710100, China; 2.  Henan Provincial Center for Disease Prevention and Control, Zhengzhou 450000, China)
 全文: PDF(5273 KB)   HTML ( ) 
摘要： 采用微生物发酵法获取天然产物,是一项比植物提取法更为经济的环境友好技术。为建立一种微生物生产苦参碱的方法,本文从陕北油田石油污染土壤中分离、筛选和鉴定了一种能合成苦参碱的枝顶孢属真菌。首先采用薄层层析法、气相色谱质谱联用法等分别确定了在所获真菌菌株发酵产物中含有苦参碱物质;使用光学显微镜观察发现,该菌株菌落呈白色绒毛状,呈圆形,菌丝分散,背面奶油色至赭黄色;菌丝无色,光滑且具隔膜,孢子囊梗直接从菌丝上长出,呈锥形,多单生;分生孢子无色,光滑,形成簇状或链状,长纺锤形,两头尖,未见厚垣孢子。内转录区间序列(internal transcribed sequence,ITS)扩增、测序后进行同源性比较发现,该菌株ITS序列与枝顶孢属的同源性高达99%。构建系统发育树表明,该真菌与枝顶孢属菌的遗传距离最近。综合形态学和遗传进化分析结果对此菌株进行了种属分类,确认这株能合成苦参碱物质的真菌应归属子囊菌门(Ascomycota)的枝顶孢属(Acremonium sp.)。
Abstract: e increasing market demand. The production of matrine by microbial fermentation, however, would be an alternative for the production obtained from plants, and must have more advantages over the extracting from plants, with less labor and time consumed. However, there are no many reports concerning microbes with capacity to produce matrine. Hence, it is very necessary to obtain matrine-producing microorganisms for establishing the bio-production by microbial fermentation. The study presented hereby was carried out to isolate and identify the fungus with the ability to synthesize matrine from petroleum-contaminated soil samples. The fungal isolate with the capability to synthesize matrine was classified by morphological and molecular biological methods. The fungal isolates were firstly obtained from samples of oil-contaminated soil in North Shaanxi, China, through spread-plate technique and plate streaking. The chemical compositions in fermentation broth of the obtained fungus were detected by thin-layer chromatography and gas chromatograph-mass spectrometer. Subsequently, the isolate was observed under the light microscope and the classification of the isolate was performed preliminarily according to the morphological characteristics. The internal transcribed sequence (ITS) of the isolated fungus was amplified and sequenced. Homology comparison was then conducted and the phylogenetic tree was established based on the sequences of other strains from previous reports. Finally, the matrine-producing fungus was classified according to the combined results of both morphology observation and phylogenetic analysis. The experimental results showed that a fungal strain was isolated from the samples of petroleum-contaminated soil. The results of both thin-layer chromatography and gas chromatograph-mass spectrometer indicated that the fungus had the capacity to produce matrine. The morphological observation showed that the colony of this fungus was white villous, circular, abaxially cream to reddish brown, with mycelia scattered. The hyphae was colorless, smooth and septate. The sporangiophores were conical, mostly solitary, showing to grow directly from the hyphae. The conidia were colorless, smooth, showing clusters or chains, with long spindle shaped, pointed at both ends and there was no chlamydospore observed. These morphological characteristics of this fungal strain were highly similar to that of Acremonium sp. and Simplicillium sp. reported previously. The results of the fungal homology comparison showed that the fungus was more closely related to Acremonium sp. with 99% homology and the phylogenetic analysis indicated that there was no significant difference in genetic distance between this strain and Acremonium. Integrating these above results, the fungus should belong to Acremonium, with the name of Acremonium sp. P0997 in this study. As there are not so many reports about Acremonium sp. to produce matrine, our study has provided much information about this species with the capacity to produce matrine. These results would contribute to establishing matrine production by microbes. Therefore, this study concerning for Acremonium sp. with the capacity to synthesize matrine has a promising application prospect for matrine production by microbes. Further researches are ongoing about the metabolites of the genus in our laboratory. The study would provide an important scientific basis for establishing matrine production by microorganisms.
出版日期: 2015-09-20
CLC:  X 13  
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
毛东霞
郭丹丹
吴玲玲
田晓雪
张胜翔
刘陶
马小魁

引用本文:

毛东霞, 郭丹丹, 吴玲玲, 田晓雪, 张胜翔, 刘陶, 马小魁. 石油污染土壤中产苦参碱真菌的分离与鉴定[J]. 浙江大学学报(农业与生命科学版), 2015, 41(5): 586-592.

Mao Dongxia, Guo Dandan, Wu Lingling, Tian Xiaoxue, Zhang Shengxiang, Liu Tao, Ma Xiaokui. Isolation and identification of a matrine-producing fungal strain from petroleum-contaminated soil. Journal of Zhejiang University (Agriculture and Life Sciences), 2015, 41(5): 586-592.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2015.03.031    或    http://www.zjujournals.com/agr/CN/Y2015/V41/I5/586

[1] 谷成刚，相雷雷，任文杰，吴为，刘畅，方国东，王芳. 土壤中酞酸酯多界面迁移转化与效应研究进展[J]. 浙江大学学报(农业与生命科学版), 2017, 43(6): 700-712.
[2] 张薇，施加春. 不同钝化剂与蚯蚓联合处理对猪粪堆肥中铜锌的钝化效果[J]. 浙江大学学报(农业与生命科学版), 2017, 43(6): 775-786.
[3] 刘晨，郭佳，赵敏，钟斌，郭华，侯淑贞，徐炜杰，杨芸，王任远，叶正钱，柳丹. 毛竹幼苗与伴矿景天间作对铜、镉、锌转运积累的影响[J]. 浙江大学学报(农业与生命科学版), 2017, 43(5): 615-622.
[4] 张栋，刘兴元，赵红挺. 生物质炭对土壤无机污染物迁移行为影响研究进展[J]. 浙江大学学报(农业与生命科学版), 2016, 42(04): 451-459.
[5] 兰天，楚颖超，张玲玲，赵文，潘运舟，张家玮，朱治强，吴蔚东. 椰衣和椰壳生物质炭的制备及其对溶液中Pb2+的吸附[J]. 浙江大学学报(农业与生命科学版), 2016, 42(04): 469-477.
[6] 陈方, 张晓燕, 黄平捷, 侯迪波, 张光新, 赵佳佳, 何纪平. 饮用水有机污染物的三维荧光光谱检测与分析方法[J]. 浙江大学学报(农业与生命科学版), 2016, 42(3): 368-377.
[7] 徐秋桐1, 常跃畅1， 章明奎1,2*. 浙江省土壤中细菌对抗生素耐药性的生态分布特征[J]. 浙江大学学报(农业与生命科学版), 2013, 39(5): 537-544.
[8] . 矿物改良剂对污染土壤中重金属多目标有效性的影响[J]. 浙江大学学报(农业与生命科学版), 2012, 38(5): 629-638.