白术中一株植物促生细菌的分离鉴定及培养条件优化  

A plant growth-promoting bacterium in Atractylodes macrocephala Koidz:isolation,identification,and optimization of culture conditions

作　　者：何勇 许淑雅 胡贤君 卢云亮 刘幸 梁倩文 齐永霞[1] 陈方新[1] HE Yong;XU Shuya;HU Xianjun;LU Yunliang;LIU Xing;LIANG Qianwen;QI Yongxia;CHEN Fangxin(Key Laboratory of Integrated Pest Management on Crops of Anhui Province,Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutes,School of Plant Protection,Anhui Agricultural University,Hefei 230031,Anhui,China;College of Agronomy,Anhui Agricultural University,Hefei 230031,Anhui,China)

机构地区：[1]安徽农业大学植物保护学院作物有害生物综合治理安徽省重点实验室植物病虫害生物学与绿色防控安徽普通高校重点实验室,安徽合肥230031 [2]安徽农业大学农学院,安徽合肥230031

出　　处：《微生物学通报》2023年第9期3983-3996,共14页Microbiology China

基　　金：中国烟草总公司贵州省公司科技项目(110202202014)。

摘　　要：【背景】白术是十分重要的大宗药材之一,在其种植过程中一直受到土壤肥力不足导致农药化肥大量施用的困扰。【目的】从白术中分离纯化出对白术生长发育具有促进作用的微生物,为白术生物肥料的制备提供材料基础。【方法】采用钼锑抗比色法和Salkowski比色法等测定菌株溶解无机磷和产吲哚乙酸(indole-3-acetic acid,IAA)的能力;使用灌根法测定供试菌株对植株生长发育的影响;通过形态学和生理生化的方法,结合16S rRNA基因测序技术对供试菌株进行分类鉴定;使用响应面法优化菌株的培养条件。【结果】分离获得一株对白术具有促生作用的菌株BZ-8,解磷能力和产IAA能力可达到4.89 mol/L和45.52μg/mL,16S rRNA基因测序长度1363 bp,系统发育树分析确定菌株BZ-8为粘质沙雷氏菌(Serratia marcescens);菌株发酵液对白术苗期的生长有明显的促进作用;优化后的发酵培养条件为(g/L):麦芽糖5.0,牛肉膏3.0,酵母提取物5.0,胰蛋白胨10.0,柠檬酸钠5.0,培养基初始pH 7.01,培养温度30.4℃,摇床转速180 r/min。【结论】白术内生菌粘质沙雷氏菌BZ-8能够解磷和产IAA,对白术具有明显的促生作用,本研究确定了菌株BZ-8的培养条件。[Background]Atractylodes macrocephala Koidz is an important and widely used Chinese medicinal plant,while its cultivation suffers from poor soil fertility which is often dealt with the heavy application of pesticides and fertilizers.[Objective]To provide a material basis for the preparation of a biofertilizer for A.macrocephala with the plant growth-promoting microorganisms isolated from the plant.[Methods]Mo-Sb colorimetry and Salkowski’s method were employed to determine the abilities of the bacterial strain to solubilize inorganic phosphorus and synthesize indole-3-acetic acid(IAA).The effects of the tested strain on plant growth were assessed with the root irrigation method.The strain was identified based on the morphological and physiological characteristics and the 16S rRNA gene sequence.Response surface methodology was employed to optimize the culture conditions of the strain.[Results]The capabilities of the isolate BZ-8 to solubilize inorganic phosphorus and produce IAA reached 4.89 mol/L and 45.52μg/mL,respectively.Its genome was 1363 bp long,and phylogenetic analysis identified the strain as Serratia marcescens.The fermentation broth of the strain significantly promoted the growth of A.macrocephala seedlings.The optimized culture conditions of BZ-8 were 5.0 g/L maltose,3.0 g/L beef extract,5.0 g/L yeast extract,10.0 g/L tryptone,5.0 g/L sodium citrate,initial pH 7.01,and incubation at 30.4℃and 180 r/min.[Conclusion]S.marcescens BZ-8,an endophytic bacterium of A.macrocephala,is capable of solubilizing phosphorus and producing IAA,demonstrating a significant promoting effect on the growth of A.macrocephala.The optimal culture conditions of strain BZ-8 were determined via response surface test.

关 键 词：植物内生细菌 粘质沙雷氏菌 促生作用 系统发育分析 培养条件优化 

分 类 号：S567.233[农业科学—中草药栽培]