黄连种子内生细菌的分离鉴定及功能验证  

Isolation,identification and functional verification of bacteria from seeds of Coptis chinensis

作　　者：向益青 廖海浪 李娜 钟芙蓉[1,2] 彭茂瑶 马云桐[1,2] XIANG Yi-qing;LIAO Hai-lang;LI Na;ZHONG Fu-rong;PENG Mao-yao;MA Yun-tong(State Key Laboratory of Traditional Chinese Medicine Resources with Southwest Characteristics;School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Institute of Economic Crop Breeding and Cultivation,Sichuan Academy of Agricultural Sciences,Chengdu 611300,China)

机构地区：[1]西南特色中药资源国家重点实验室 [2]成都中医药大学药学院,成都611137 [3]四川省农业科学院经济作物育种栽培研究所,成都611300

出　　处：《天然产物研究与开发》2023年第2期191-199,共9页Natural Product Research and Development

基　　金：濒危道地药材“雅连”精深加工技术及新产品(2022ZHCG0096);四川省中医药产业发展重大项目(510201202109711)。

摘　　要：植物种子具有丰富的微生物资源,为深入挖掘黄连种子促生内生菌,本研究通过平板分离获得种子可培养内生菌,对获得的菌株进行溶磷、固氮、产铁载体和产IAA功能研究,筛选具有促生特性的菌株,结合种子萌发实验进行菌株促生功能验证,并对其进行分子生物学鉴定。结果表明,黄连种子可培养内生菌丰富,共分离得到内生菌57株,经16S rRNA鉴定出内生细菌29株,主要分布于芽孢杆菌属Bacillus、寡养假单孢杆菌属Stenotrophomonas、微杆菌属Microbacterium、无色杆菌属Achromobacter等13个属。优势菌为芽孢杆菌属(7株)和寡养假单孢杆菌属(7株),分别占总鉴定菌株的24.14%。促生特性研究表明,具有溶磷功能菌株2株,固氮功能菌株9株产铁载体菌株2株,产IAA菌株6株,其IAA分泌量为64.52~203.57 mg/L。其中既具有溶磷功能又产IAA菌株1株,既溶磷又产铁载体菌株1株,既产铁载体又产IAA菌株1株。Priestia megaterium和A.spanius可显著提高黄连种子的发芽率。本研究将为揭示种子内生菌的促生机制和顽拗性种子后熟与萌发机制提供理论依据,同时也为微生物菌肥的开发奠定基础。Plant seeds are rich in microbial resources.For digging into the endophytic bacteria of seeds of Coptis chinensis,endophytic bacterial strains were isolated from seeds with plate isolation,and the obtained strains were studied on phosphate-dissolving,nitrogen fixation,siderophores and IAA production.The strains with growth-promoting properties were screened,and were tested for growth-promoting effects in combination with the seed germination experiment,as well as molecularly identified.The results showed that seeds of C.chinensis were rich in culturable endophytic bacteria.A total of 57 endophytic bacteria strains were isolated,and 29 strains were identified by 16S rRNA,mainly distributed in 13 genera such as Bacillus,Stenotrophomonas,Microbacterium and Achromobacter.The dominant bacteria were Bacillus(seven strains)and Stenotrophomonas(seven strains),accounting for 24.14%of the total identified strains respectively.The study of growth-promoting characteristics showed that there were two strains with phosphorus-dissolving function,nine strains with nitrogen-fixing function,two siderophores-producing strains,and six strains with IAA-producing function,and their IAA secretion amount ranged from 64.52-203.57 mg/L.Among them,one strain was capable of both phosphate-dissolving and IAA production,and one strain as capable of phosphate-dissolving and siderophore production,and one strain was capable of siderophore production and IAA production.Priestia megaterium and A.spanius could significantly increase the germination percentage of seeds of C.chinensis.This study will provide a theoretical basis for revealing the growth promotion mechanism of seed endophytes and understanding the mechanism of after-ripening and germination of recalcitrant seeds,and also lay a foundation for the development of microbial fertilizer.

关 键 词：黄连种子 内生细菌 物种鉴定 功能验证 

分 类 号：R284.1[医药卫生—中药学] R93[医药卫生—中医学]