天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (6): 1027-1037.doi: 10.16333/j.1001-6880.2022.6.016

• 开发研究 • 上一篇    下一篇

不同生育期襄麦冬内生细菌的分布及产甾体皂苷菌株的初步筛选

余海忠1*,程   旭3,王海燕1,殷幼平2,王中康2   

  1. 1湖北文理学院食品科学技术学院,襄阳 441053;2重庆大学生命科学学院,重庆 405200;3襄阳市农业科学院,襄阳 441057
  • 出版日期:2022-06-28 发布日期:2022-06-30
  • 基金资助:
    国家星火计划面上项目(2015GA760011);湖北省高价值知识产权培育工程(专利类)项目(D2020000875);襄阳市医疗卫生领域科技计划重点项目(2020YL23)

Distribution of endophytic bacteria and preliminary screening of steroidal saponin-producing strain from Liriope spicata var. prolifera  Y.T.Ma in different growth stages

YU Hai-zhong1*,CHENG Xu3,WANG Hai-yan1,YIN You-ping2,WANG Zhong-kang2   

  1. 1School of Food Science and Technology,Hubei University of Arts and Science,Xiangyang 441053,China;2College of Life Sciences, Chongqing University,Chongqing 405200,China;3Xiangyang Academy of Agricultural Sciences, Xiangyang 441057,China
  • Online:2022-06-28 Published:2022-06-30

摘要:

甾体皂苷是百合科山麦冬属多年生草本植物襄麦冬Liriope spicata var. prolifera Y.T.Ma的主要活性成分之一,具有多重药理学活性。为了探究不同生育期襄麦冬内生细菌的分布情况,挖掘其产甾体皂苷内生细菌菌株资源,本研究对其内生细菌进行了分离、纯化、鉴定和功能研究。利用纯培技术从不同生育期的块根和须根中分离获得50株细菌,分子生物学鉴定这些内生细菌分属15个属22个种。利用16S rDNA-PCR-DGGE技术分析襄麦冬块根始见期、迅速膨大期、成熟采收期的块根内生细菌的群落结构,结果表明三个生育期的块根样品内生菌群结构基本相同,细菌类群差异很小,获得的21个序列中包含分属14属 21种的细菌类群。利用颜色反应、TLC、HPLC等方法,从纯培细菌中初步筛选出墨西哥假黄单胞菌Pseudoxanthomonas mexicana、山羊葡萄球菌Staphylococcus caprae、砖红色微杆菌Microbacterium testaceum 3株细菌具有明显产甾体皂苷特征。上述结果表明襄麦冬内生细菌具有“工厂化”制备生产甾体皂苷的应用开发潜力。

关键词: 襄麦冬, 内生细菌, 甾体皂苷, 分布

Abstract:

Liriope spicata var.prolifera Y.T.Ma is a indigenous medicinal herb cultivated in Xiangyang of Hubei,China,which belongs to Liriope genus of Liliaceae family.Steroidal saponins,one of the main active components in the tuberous roots of L. spicata var. prolifera,have multiple pharmacological activities.In order to investigate the distribution of endophytic bacteria in L. spicata var. prolifera at different growth stages and to explore the resources of endophytic bacteria producing steroidal saponins,the endophytic bacteria were isolated,purified,identified and function studied.Using pure culture technology,50 endophytic bacteria were isolated from L. spicata var. prolifera  at different growth stages,and these endophytic bacteria were 22 species belonging to 15 genera by colony morphology and molecular biological identification.In addition,16S rDNA PCR-DGGE method was used to analyze the endophytic bacteria distribution in the tuberous root of L. spicata var. prolifera  during the different stages including initial growth stage,rapid growth stage and mature stage.DGGE showed that the structures of endophytic bacteria in tuberous root from three growing periods were all basically the same and slightly difference.By blasting the sequences of 16S rDNA V6-V8 hypervariable region in GenBank,it was found that the 21 sequences contained 21 bacteria species belonging to 14 genera.Moreover,three strains of Pseudoxanthomonas mexicana,Staphylococcus capraeand Microbacterium testaceum were preliminarily identified as possessing the steroidal saponin-producing ability by Color reaction,TLC and HPLC.The above results showed that the endophytic bacteria of L. spicata var. prolifera have the application and development potential of "industrialized" preparation and production of steroidal saponins.

Key words: Liriope spicata var. prolifera Y.T.Ma, endophytic bacteria, steroidal saponins, distribution

中图分类号:  Q936