生物技术通报 ›› 2017, Vol. 33 ›› Issue (9): 85-93.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0251

• 技术与方法 • 上一篇    下一篇

PCR-DGGE技术监测自然发酵葛根中微生物菌群动态变化

冉淦侨1,3,张红艳2,任平1   

  1. 1. 陕西省科学院酶工程研究所,西安 710600;
    2. 陕西省酶工程技术中心,西安 710600;
    3. 陕西省生物农业研究所,西安 710600
  • 收稿日期:2017-03-30 出版日期:2017-09-01 发布日期:2017-09-15
  • 作者简介:冉淦侨,女,硕士,助理研究员,研究方向:植物保护;E-mail:ranganqiao@163.com
  • 基金资助:
    陕西省科学院青年人才培养专项(2012k-31)

Dynamic Changes of Microbial Community in the Process of Natural Fermentation of Puerariae lotaba Monitored by PCR-DGGE

RAN Gan-qiao1,3,ZHANG Hong-yan2,REN Ping1   

  1. 1. Institute of Enzyme Engineering,Shaanxi Academy of Science,Xi’an 710600;
    2. Enzyme Engineering Technology Center of Shaanxi Province,Xi’an 710600;
    3. Shaanxi Bio-Agriculture Institute,Xi’an 710600
  • Received:2017-03-30 Published:2017-09-01 Online:2017-09-15

摘要: 采用自然堆积发酵对葛根进行预处理,处理后葛根总异黄酮的提取量提高了5.7% 左右。随后,分别采用传统平板法和PCR-DGGE技术,对发酵过程中不同时期微生物菌群的变化进行分析,从而筛选出发酵中起主要作用的微生物。结果表明,自然发酵过程中的细菌呈升高-降低-再升高的变化趋势:升温期明显增多,高温期有所下降,降温期再次增多;真菌的数量在整个发酵过程中都呈持续升高的趋势。发酵过程中起主要作用的微生物也被初步确定,它们分别是地衣芽孢杆菌、桃色欧文氏菌、芽孢杆菌属、枯草芽孢杆菌、不可培养细菌、爪哇青霉、黑曲霉、溜曲霉、米根霉和不可培养曲霉。

关键词: 自然发酵, 微生物群落, 总异黄酮, 变性梯度凝胶电泳(DGGE)

Abstract: In this study,the extraction yield of total isoflavones from Puerariae lotaba increased by 5.7% after pretreated with natural compost fermentation. After that,in order to determine the microorganisms playing a major role in this fermentation,the dynamic changes of the microbes in the process of natural fermentation of P. lotaba powder were analyzed using traditional flat culture method and PCR-DGGE technology respectively. The results showed that the number of bacteria showed an “increase-decrease-increase” trend in the whole fermentation process:increased significantly in the temperature-rising stage,then decreased slightly in the thermophilic stage,and finally rose again in the temperature-cooling stage,but the number of fungi presented a continuous increase trend. Besides,several microorganisms isolated from P. lotaba were initially identified to have positive effects on improving the extraction yield of total isoflavones,and these microbes included:Bacillus licheniformis,Rhizoctonia solani,Bacillus,Bacillus subtilis,non-cultivable bacteria,Penicillium japonicum,Aspergillus niger,Aspergillus,Asparagus,and non-cultivable Aspergillus.

Key words: natural fermentation, microbial community, total isoflavones, denatured gradient gel electrophoresis(DGGE)