艾草根系DSE菌株与根际功能性细菌的分离、鉴定及应用

doi:10.12190/j.issn.2096-1197.2024.03.05

摘要/Abstract

摘要： 【目的】从艾草（Artemisia argyi）根系及根际土壤中分离筛选具有促生作用的菌种。【方法】分别在内蒙古鄂尔多斯市和通辽市采集艾草根系及根际土壤样本,分离纯化共生真菌——深色有隔内生真菌（dark septate endophytes,DSE）和功能性细菌,采用分子生物学技术进行鉴定,从中筛选具有显著促生效果的DSE菌种和功能性细菌,构建人工菌群,并设置不接菌（A）、单接DSE（B）、单接解磷菌（C）、单接固氮菌（D）、DSE+解磷菌（E）、DSE+固氮菌（F）、解磷菌+固氮菌（G）、DSE+解磷菌+固氮菌（H）8个处理对艾草植株进行回接试验,以筛选最佳菌种。【结果】DSE真菌在艾草根系中的定殖率最高,共生结构主要呈现为链状、条带状、脑状和零散状。在艾草根系中,成功分离出4株DSE,其中菊异茎点霉（Paraphoma chrysanthemicola）在两试验地均为优势菌种。在艾草根际土壤中,共分离出36株功能性细菌,分别为26株解磷细菌和10株固氮细菌。解磷细菌中以芽孢杆菌属（Bacillus）为主,占总分离数的30.50%,其中,短小芽孢杆菌（Bacillus pumilus）占比为27.20%;固氮细菌中以根瘤菌属（Rhizobium）为主,占总分离数的16.67%,其中,溶纤维根瘤菌（Rhizobium cellulosilyticum）占比为33.30%。B~H处理的艾草生长指标均优于A处理,其中,H处理的促生效果最佳,其茎粗、叶片数、地上生物量、地下生物量分别提高了24.93%、80.00%、279.17%和251.69%。【结论】异茎点霉（Paraphoma chrysanthemicola）、短小芽孢杆菌（Bacillus pumilus）和溶纤维根瘤菌（Rhizobium cellulosilyticum）复合接种的促生效果最佳,DSE+解磷菌+固氮菌处理为最佳促生微生物组合。

关键词: 艾草根系, 根际土壤, 深色有隔内生真菌（DSE）, 功能性细菌, 分离, 鉴定

Abstract: 【Objective】To isolate and screen plant growth-promoting strains from the roots and rhizosphere soil of Artemisia argyi.【Methods】To collect root and rhizosphere soil samples of Artemisia argyi from Ordos and Tongliao in Inner Mongolia,isolate and purify symbiotic fungi-dark septate endophytes（DSE） and functional bacteria,identify them using molecular biology techniques,screen DSE strains and functional bacteria with significant growth promoting effects,construct artificial microbial communities,and set up non inoculated bacteria（A）,single inoculated DSE（B）,single inoculated phosphate-solubilizing bacteria（C）,single inoculated nitrogen-fixing bacteria（D）,DSE+phosphate-solubilizing bacteria（E）,DSE+nitrogen-fixing bacteria（F）,phosphate-solubilizing bacteria+nitrogen-fixing bacteria（G）,DSE+phosphate-solubilizing bacteria+nitrogen-fixing bacteria（H） eight treatments were used to conduct a rewinding experiment on Artemisia argyi plants to screen for the best bacterial strain.【Results】The colonization rate of DSE fungi in the roots of Artemisia argyi was the highest,and the symbiotic structure was mainly chain,strip,brain and scattered. Four strains of DSE were successfully isolated from the roots of Artemisia argyi,among which Paraphoma chrysanthemicola was the dominant strain in both test sites. A total of 36 functional bacteria were isolated from the rhizosphere soil of Artemisia argyi,including 26 phosphate-solubilizing bacteria and 10 nitrogen-fixing bacteria. The phosphate-solubilizing bacteria were mainly Bacillus,accounting for 30.50% of the total number of isolates,of which Bacillus pumilus accounted for 27.20%. Rhizobium was the main nitrogen-fixing bacteria,accounting for 16.67% of the total number of isolates,of which Rhizobium cellulosilyticum accounted for 33.30%.The growth indexes of Artemisia argyi in B-H treatments were better than those in A treatment. Among them,H treatment had the best growth-promoting effect,and its stem diameter,leaf number,aboveground biomass and underground biomass increased by 24.93%,80.00%,279.17% and 251.69%,respectively.【Conclusion】The compound inoculation of Paraphoma chrysanthemicola,Bacillus pumilus and Rhizobium cellulosilyticum had the best growth-promoting effect,and DSE+phosphate-solubilizing bacteria+nitrogen-fixing bacteria treatment was the best growth-promoting microbial combination.

Key words: Artemisia argyi root system, Rhizosphere soil, Dark septate endophytic, Functional bacteria, Isolation, Identification

中图分类号:

S567.23

乌有汗, 曹露彦, 许明慧, 欧阳瑞培, 石俊庭, 姚庆智. 艾草根系DSE菌株与根际功能性细菌的分离、鉴定及应用[J]. 北方农业学报, 2024, 52(3): 49-56.

Wuyouhan, CAO Luyan, XU Minghui, OUYANG Ruipei, SHI Junting, YAO Qingzhi. Isolation,identification,and application of DSE strain from Artemisia argyi root system and functional bacteria in the rhizosphere[J]. Journal of Northern Agriculture, 2024, 52(3): 49-56.

参考文献

[1] BATIHA G E S,OLATUNDE A,EL-MLEEH A,et al.Bioactive compounds,pharmacological actions,and pharmacokinetics of wormwood(Artemisia absinthium)[J].Antibiotics,2020,9(6):353.
[2] 左红娟,曹辉,张晓申.艾草综合开发利用研究现状、存在问题及发展展望[J].特种经济动植物,2023,26(3):145-147.
[3] 张新学. 西北旱作区艾草绿色增效栽培技术[J].农业科技与信息,2023,20(10):27-30.
[4] 何立威,付晨青,王秀萍,等.我国艾草标准化栽培技术及加工应用研究进展[J].安徽农业科学,2021,49(16):4-6.
[5] XIE J,LUO C Q,YANG X W,et al.Study on wild medicinal plant resources and their applied ethnology in multiethnic areas of the Gansu-Ningxia-Inner Mongolia intersection zone[J].Journal of Ethnobiology and Ethnomedicine,2023,19(1):18.
[6] 刘素,吴宏亮,陈倬,等.连作年限影响芹菜根际土壤微生物群落结构及功能类群[J].中国农业气象,2023,44(5):372-385.
[7] 冯路遥,赵江源,施竹凤,等.森林根际土壤细菌的分离、鉴定及生物活性筛选[J].生物技术通报,2024,40(1):294-307.
[8] 慕瑞瑞,翟雪宁,李海健.深色有隔内生真菌对番茄种植土壤的改良效果研究[J].乡村科技,2023,14(15):79-81.
[9] 闫巧芝,李晴,苗志加,等.深色有隔内生真菌(DSE)功能和重金属耐性机制研究进展[J].农业环境科学学报,2024,43(2):242-253.
[10] NETHERWAY T,BENGTSSON J,BUEGGER F,et al.Pervasive associations between dark septate endophytic fungi with tree root and soil microbiomes across europe[J].Nature Communications,2024,15:159.
[11] 张小龙,王嘉瑞,李青卓,等.合成微生物群落及在发酵食品中应用研究进展[J].中国酿造,2021,40(3):17-21.
[12] 刘燕霞,龙俊萌,王静茹,等.五种漠境深色有隔内生真菌对小麦促生抗旱效应研究[J].中国科学:生命科学,2021,51(9):1339-1349.
[13] 孟超楠,赵玉洁,陈佳欣,等.2株青贮玉米根际固氮菌的筛选鉴定及促生作用研究[J].草业学报,2024,33(3):174-185.
[14] 周童晖,卜建超,张皓珊,等.巨菌草根际高效解磷菌的筛选[J].福建农林大学学报(自然科学版),2023,52(1):26-32.
[15] 冯丹. 款冬根系DSE的多样性及其与PGPR的接种效应[D].太原:山西师范大学,2021.
[16] 东美奇,肖宇峰,王思雅,等.复合菌YM4对烟嘧磺隆降解及对土壤微生物影响[J].农药,2024,63(1):32-38.
[17] 谭艺. DSE真菌促进蓝莓磷吸收的影响机制研究[D].贵阳:贵州大学,2023.
[18] 熊珂敏,陈嘉欣,荣世炎,等.云南典型铅锌矿区植物根部AMF和DSE侵染特征研究[J].湖南生态科学学报,2023,10(3):1-9.
[19] 陈思杰,李金,杜娟,等.枸杞根系内生真菌的染色方法[J].河北大学学报(自然科学版),2021,41(4):406-411.
[20] 欧阳瑞培. 内蒙古艾蒿丛枝菌根真菌和深色有隔内生真菌的研究[D].呼和浩特:内蒙古农业大学,2021.
[21] 欧阳瑞培,姚庆智,石俊庭,等.艾蒿根系共生真菌与土壤理化因子的相关性分析[J].北方农业学报,2021,49(2):49-55.
[22] 汤子祥. 艾草根系促生菌的筛选鉴定及其应用效果研究[D].杨凌:西北农林科技大学,2022.
[23] 赵义. 番茄根际促生菌的筛选鉴定及其功能研究[D].福州:福建农林大学,2022.
[24] 张瑞福,沈其荣.微生物肥料新型功能作用机理与根际定殖增强策略[J].微生物学杂志,2024,44(1):1-11.
[25] 栗瑞红,李红霞,李长胜,等.微生物菌剂对连作蒙古黄芪生物量及土壤真菌群落结构的影响[J].北方农业学报,2022,50(6):50-56.
[26] 宋志远,聂波.高效解磷菌的筛选及应用研究[J].现代农业研究,2020,26(11):56-57.
[27] 杨晓蕾,李建宏,姚拓,等.复合促生菌剂发酵条件优化及其对青稞促生效果评价[J].草地学报,2022,30(1):212-219.
[28] 张海珠,李杨,张彦如,等.菌根真菌处理下滇重楼对营养元素的吸收和积累[J].环境化学,2019,38(3):615-625.
[29] 石俊庭. 内蒙古艾草种质资源的评价和不同微生物组合对艾草产量及质量的影响[D].呼和浩特:内蒙古农业大学,2022.
[30] 包雯静. 多功能合成微生物群落YJ-1的构建及在土壤修复的应用研究[D].哈尔滨:东北农业大学,2023.