森林根际土壤细菌的分离、鉴定及生物活性筛选  

Isolation and Identification of Bacteria in Forest Rhizosphere Soil and Their Biological Activity Screening

作　　者：冯路遥 赵江源[3] 施竹凤 莫艳芳 杨童雨 申云鑫 何飞飞[1] 李铭刚[3] 杨佩文[2] FENG Lu-yao;ZHAO Jiang-yuan;SHI Zhu-feng;MO Yan-fang;YANG Tong-yu;SHEN Yun-xin;HE Fei-fei;LI Ming-gang;YANG Pei-wen(Resource Plant Research Institute,Yunnan University,Kunming 650504;Institute of Agricultural Environment and Resources,Yunnan Academy of Agricultural Sciences,Kunming 650205;Yunnan Institute of Microbiology,Yunnan University,Kunming 650504;College of Plant Protection,Yunnan Agricultural University,Kunming 650201)

机构地区：[1]云南大学资源植物研究院,昆明650504 [2]云南省农业科学院农业环境资源研究所,昆明650205 [3]云南大学云南省微生物研究所,昆明650504 [4]云南农业大学植物保护学院,昆明650201

出　　处：《生物技术通报》2024年第1期294-307,共14页Biotechnology Bulletin

基　　金：云南省科技计划重大科技专项(202202AE090015,202202AE090010);云南省专家基层科研工作站-杨佩文专家工作站(嵩明县)(云人社通「2022」17号);云南省专家基层科研工作站-杨佩文专家工作站(陆良县)(云人社通「2022」17号)。

摘　　要：【目的】从无量山国家级自然保护区森林根际土壤发掘具有多种生物活性的功能菌株,探究其开发应用潜力。【方法】采集无量山地区25个区域植物的根际土壤,采用选择培养基,分离鉴定磷酸盐溶解、固氮、溶锌和拮抗等活性菌株,进一步测定菌株分泌铁载体、ACC脱氨酶和吲哚乙酸等生物活性,并验证促番茄种子发芽和生长效果。【结果】分离鉴定得到解磷菌70株,固氮菌27株,解钾菌8株,拮抗镰刀菌的菌株51株。其中,YIM B08401和YIM B08402形态学结合生理生化特性和16S rDNA序列测序,鉴定为白色伯克霍尔德氏菌(Burkholderia alba)和青岛假单胞菌(Pseudomonas qingdaonensis),两个菌株均具有磷酸盐溶解、固氮、溶锌和分泌铁载体的活性,最大可溶性磷含量为(455.63±59.65)mg/L和(878.95±64.78)mg/L;两株菌的促种子发芽试验结果接近,施加稀释10倍、10^(2)倍和10^(3)倍的发酵上清液后,发芽率都维持在82%-93%,明显高于对照组的56%和49%,施加菌株发酵液的处理组相较于空白对照组的长度都有显著增加。盆栽实验证明,两株菌株促生效果最明显的处理组在地上部长度、鲜重、干重、茎粗、根长、根鲜重、根干重方面的数据都显著优于对照组,YIM B08401的上述指标相对于对照组分别显著增加了89%、495%、268%、62%、53%、385%和469%,YIM B08402的上述指标相对于对照组分别显著增加了118%、528%、477%、55%、37%、413%和747%。此外,菌株YIM B08401还具有拮抗病原菌和分泌ACC脱氨酶活性,YIM B08402则还具有分泌吲哚乙酸的活性。【结论】无量山森林根际土壤蕴含具有多种生物活性的功能菌株,白色伯克霍尔德氏菌(B. alba, YIM B08401)和青岛假单胞菌(P. qingdaonensis, YIM B08402)具备开发为新型微生物肥料的应用潜力。【Objective】Functional strains with multiple biological activities were excavated from the forest rhizosphere soil of Wuliangshan National Nature Reserve,and their potential for development and application was explored.【Method】The 25 rhizosphere soil samples from different regions and plants in Wuliangshan National Nature Reserve were collected.The active strains with functions such as phosphate dissolution,nitrogen fixation,zinc dissolution,and antibacterial activity were isolated and identified using selective culture medium.The biological activities of these strains,such as secreted siderophore,ACC deaminase,and indoleacetic acid,were tested while verifying their effectiveness in promoting tomato seed germination and plant growth.【Result】70 phosphorus-dissolving bacteria,27 nitrogen-fixing bacteria,8 potassium-dissolving bacteria,and 51 antibacterial bacteria were isolated and identified.Among them,YIM B08401 and YIM B08402 were identified as Burkholderia alba and Pseudomonas qingdaonensis based on morphological,physiological,biochemical characteristics,and 16S rDNA sequencing.The two strains both had the activities of phosphate lysis,nitrogen fixation,zinc lysis and siderophore secretion.Their maximum soluble phosphorus content was(455.63±59.65)mg/L and(878.95±64.78)mg/L.The results of seed germination promotion tests of the two strains were similar,and the germination rates of the two strains were maintained at 82%-93%after applying the fermentation 23 supernatant diluted 10 times,10^(2) times and 10^(3) times,which was significantly higher than 56%and 49%in the control group.The length of the treatment group with the fermentation liquid of the strain significantly increased compared with the blank control group.Pot experiment showed that the above ground length,fresh weight,dry weight,stem diameter,root length,fresh weight and dry weight of the two strains in the treatment group with the most obvious growth promotion effect were significantly better than that in the control group.Compared with

关 键 词：无量山森林根际土壤 可培养微生物 白色伯克霍尔德氏菌 青岛假单胞菌 养分转化 拮抗活性 生物肥料 

分 类 号：S714.3[农业科学—林学]