文献类型：期刊文献

中文题名：茄腐镰刀菌胁迫下甘肃渭源当归根际土壤微生物及代谢物变化

英文题名：Changes in rhizosphere soil microorganisms and metabolites of Angelica sinensis in Weiyuan,Gansu under Fusarium solani stress

作者：谢田朋[1,2];张佳宁[1,2];杨林桦[1,2];丁琦[1,2];杜晓雪[1,2];刘雪枫[1,2];朱田田[1,2];杨涛[3]

第一作者：谢田朋

机构：[1]甘肃中医药大学药学院,甘肃兰州730000;[2]甘肃中医药大学陇药产业创新研究院,甘肃兰州730000;[3]甘肃省科学院生物研究所/甘肃省微生物资源开发重点实验室,甘肃兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2025

卷号：56

期号：9

起止页码：2697

中文期刊名：南方农业学报

外文期刊名：Journal of Southern Agriculture

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金项目(82160714);甘肃省青年人才项目(2024QNTD36);甘肃省高校教师创新基金项目(2025A-101);甘肃省中药质量与标准研究重点实验室开放基金项目(ZYZL-2024-04)。

语种：中文

中文关键词：当归;根腐病;高通量测序;液相色谱—质谱联用技术;茄腐镰刀菌

外文关键词：Angelica sinensis;root rot;high-throughput sequencing;liquid chromatography-mass spectrometry;Fusarium solani

摘要：【目的】探明引发甘肃渭源当归根腐病的关键致病菌,研究致病菌对当归根际土壤微生物及代谢物的影响,为当归根腐病的生防菌剂研发提供理论基础。【方法】以甘肃渭源当归为研究对象,采用组织分离法分离纯化根腐病病原菌,运用形态学及分子生物学方法对病原菌进行种类鉴定;通过盆栽试验确定病原菌对当归植株的致死浓度,选择最佳菌液浓度建立患病组模型;采用16S rDNA和ITS rDNA高通量测序技术及液相色谱—质谱联用技术(LC-MS)对病原菌胁迫下当归根际土壤微生物及代谢物变化进行分析。【结果】从患病当归根部组织分离及纯化得到10株菌株,经柯赫氏法则验证菌株Z为当归根腐病的病原菌。结合菌株Z的形态学特征和分子生物学特性,确定菌株Z为茄腐镰刀菌(Fusarium solani)。在菌株Z作用下,当归根际土壤微生物群落结构在属分类水平上发生明显变化,其中,在健康组与患病组间有29个细菌属存在极显著组间差异(P<0.01,下同),29个细菌属中除Gemmatimonadaceae_unclassified外均为2组中的非优势菌属;有7个真菌属存在极显著组间差异,且全部为患病组的优势菌属。患病组与健康组间存在14种有显著差异(P<0.05,下同)的根际土壤代谢物,其中牛磺酸(Taurine)、喹啉酸(Quinolinic acid)、6-磷酸葡萄糖酸(6-Phosphogluconic acid)和氧代己二酸(Oxoadipic acid)的含量在患病组中显著上调。对当归根际土壤中极显著差异微生物和显著差异代谢物的Pearson相关分析结果显示,在患病组中显著上调的牛磺酸、喹啉酸、6-磷酸葡萄糖酸和氧代己二酸与Blfdi19、KD3-10、Luteolibacter、糖单胞菌属(Saccharimonas)、Aquicella、Arenimonas、Ferruginibacter、丰祐菌属(Opitutus)、A4b、成对杆菌属(Dyadobacter)等10个细菌属分别存在显著或极显著正相关,而与差异真菌间不显著相关(P>0.05)。【结论】茄腐镰刀菌是导致甘肃渭源当归根腐病的关键致病菌,在其干扰下,当归根际土壤微生物群落结构和代谢物均产生明显变化,致病菌对真菌属结构的影响较细菌属强,但差异代谢物则主要与差异细菌属存在显著或极显著相关。牛磺酸、喹啉酸、6-磷酸葡萄糖酸和氧代己二酸可能是潜在的“呼救”物质,Luteolibacter、糖单胞菌属、Arenimonas、Ferruginibacter、丰祐菌属、A4b、成对杆菌属可能是被招募的潜在有益菌。
【Objective】This study aimed to identify the key pathogens causing root rot in Angelica sinensis from Weiyuan,Gansu,and to study its effects on the rhizosphere soil microorganisms and metabolites of Angelica sinensis,so as to provide a theoretical basis for research and development of biocontrol agents for root rot of Angelica sinensis.【Method】Angelica sinensis from Weiyuan,Gansu was used as the research subjects.The pathogens causing root rot were isolated and purified using tissue separation method,and they were identified using morphological and molecular biological methods.The lethal concentrations of the pathogens were determined through pot experiments,and the disease model was established using the optimal solution concentration.The changes in the rhizosphere soil microorganisms and metabolites of Angelica sinensis under the stress of pathogen were analyzed using 16S rDNA,ITS rDNA high-throughput sequencing technology,and liquid chromatography–mass spectrometry(LC-MS)methods.【Result】Ten strains were isolated and purified from root tissues of diseased Angelica sinensis.Among which,the strain Z was identified as the pathogen.Based on the morphological characteristics and molecular biological features of strain Z,it was identified as Fusarium solani.Under the effect of strain Z,the rhizosphere soil microbial community structure of Angelica sinensis changed significantly at the genus level:29 bacterial genera showed extremely significant inter-group differences between the healthy group and diseased group(P<0.01,the same below),and they were non-dominant genera in the two groups except for Gemmatimonadaceae_unclassified;7 fungal genera showed extremely significant inter-group differences,and all of them were dominant genera in the diseased group.There were 14 significantly different rhizosphere soil metabolites between the diseased group and the healthy group(P<0.05,the same below),and the contents of taurine,quinolinic acid,6-phosphogluconic acid,and oxoadipic acid were significantly up-regulated in the diseased group.Through Pearson correlation analysis of differential microorganisms at P<0.01 and differential metabolites in rhizosphere soil of Angenlica sinensis showed that the significantly up-regulated taurine,quinolinic acid,6-phosphogluconic acid,and oxoadipic acid in the diseased group had significant or extremely significant positive correlations with 10 bacterial genera,including Blfdi19,KD3-10,Luteolibacter,Saccharimonas,Aquicella,Arenimonas,Ferruginibacter,Opitutus,A4b,and Dyadobacter,but there were no significant correlations among the differential fungi(P>0.05).【Conclusion】Fusarium solani is the key pathogen causing root rot in Angelica sinensis from Weiyuan,Gansu.Interference of Fusarium solani leads to obvious changes in rhizosphere soil microbial community structure and metabolites of Angelica sinensis.The impact of the pathogen is stronger on the fungal genus structure than on the bacterial genus structure,but the differential metabolites have significant or extremely significant correlations with the differential bacterial genera.Taurine,quinolinic acid,6-phosphogluconic acid,and oxoadipic acid may be potential“cry for help”substances;Luteolibacter,Saccharimonas,Arenimonas,Ferruginibacter,Opitutus,A4b,and Dyadobacter may be potential beneficial bacteria recruited.