摘要
干腐病是浙贝母(Fritillaria thunbergii Miq.)的主要病害之一,主要由镰刀属真菌尖孢镰刀菌(Fusarium oxysporum)引起。本研究旨在从浙江省道地药材浙贝母植株中鉴定筛选潜在生防菌和化学药剂,为干腐病生物防治提供新策略。通过核糖体内转录间隔区(ITS)等6个DNA序列的PCR扩增与测序,结合GenBank同源性比对及系统发育树构建,精确鉴定了3株潜在生防真菌:钩状木霉(Trichoderma hamatum)、产红青霉(Penicillium rubens)、白囊耙齿菌(Irpex lacteus)。采用平板对峙法和琼脂稀释法评估三者与实验室现有潜在生防细菌水生拉恩氏菌(Chryseobacterium aquaticum)、枯草芽孢杆菌(Bacillus subtilis)和解淀粉芽孢杆菌(B.licheniformis)对尖孢镰刀菌的室内抑制效果。结果显示,接种5 d后,6种生防菌对尖孢镰刀菌平均抑菌率五点对峙试验分别为76.48%、50.17%、58.28%、48.88%、66.26%和23.86%;两点对峙试验分别为47.92%、50.72%、34.22%、21.07%、17.18%和13.16%。琼脂稀释法揭示,当无菌发酵滤液体积分数在1%~20%时,6种生防菌的抑制效果随浓度增加而提高,最大抑制率分别为22.91%、21.49%、23.61%、46.06%、3.93%和2.67%。化学药剂筛选结果显示,百菌清、氰烯菌酯、吡唑醚菌酯和咯菌腈抑制孢子萌发的EC_(50)值分别为1.14、4.92、0.15和0.25μg/mL,抑制菌丝生长的EC_(50)值分别为3.60、11.07、4.02和0.13μg/mL。研究结果表明,6种生防菌均可显著抑制尖孢镰刀菌,其中钩状木霉、产红青霉、白囊耙齿菌和水生拉恩氏菌展现出高效生防潜力,化学杀菌剂中咯菌腈抑菌活性最佳。
Dry rot,predominantly caused by the Fusarium genus fungus Fusarium oxysporum,is a major disease affecting Fritillaria thunbergii Miq.,an important medicinal plant in Zhejiang Province.The aim of this study was to select potential biocontrol fungi and chemical agents from indigenous medicinal plants in Zhejiang,with the goal of developing innovative biological control measures against dry rot.Utilizing PCR amplification and sequencing of six gene sequences,including the internal transcribed spacer(ITS)region,coupled with BLAST homology comparison and phylogenetic tree construction,three potentially antagonistic fungi were accurately identified:Trichoderma hamatum,Penicillium rubens,and Irpex lacteus.Assessment of the in vitro antifungal efficacy of potentially antagonistic fungi and existing laboratory strains Chryseobacterium aquaticum,Bacillus subtilis,and B.licheniformis against F.oxysporum using the agar confrontation method and the agar dilution method.The results from the five-point confrontation test showed that five days after inoculation,the average inhibition rates of the six potentially antagonistic microorganisms were 76.48%,50.17%,58.28%,48.88%,66.26%,and 23.86%,respectively.In the two-point confrontation test,the average inhibition rates were 47.92%,50.72%,34.22%,21.07%,17.18%,and 13.16%,respectively.The agar dilution method demonstrated that the inhibitory effects of the six potentially antagonistic microorganisms increased with the concentration of sterile fermentation broth within the range of 1%to 20%,with maximum inhibition rates of 22.91%,21.49%,23.61%,46.06%,3.93%,and 2.67%,respectively.The chemical fungicide screening results showed that the EC_(50) values of chlorothalonil,phenamacril,pyraclostrobin,and fludioxonil against F.oxysporum were 1.14,4.92,0.15,0.25μg/mL(spore germination)and 3.60,11.07,4.02,0.13μg/mL(mycelial growth),respectively.The results indicated that six biocontrol agents showed significant inhibitory activity,particularly T.hamatum,P.rubens,I.lacteus,and C.aquaticum.Fludioxonil exhibited the best antifungal activity among the chemical fungicides tested.
作者
严紫焰
胡光锦
赵可欣
黄垲焯
杨佳骏
赵伟春
YAN Ziyan;HU Guangjin;ZHAO Kexin;HUANG Kaizhuo;YANG Jiajun;ZHAO Weichun(School of Life Science,Zhejiang Chinese Medical University,Hangzhou 310053,China)
出处
《农药学学报》
北大核心
2025年第3期533-542,共10页
Chinese Journal of Pesticide Science
基金
浙江省基础公益研究计划(No.LGN21C140004)
国家级大学生创新创业训练计划项目(No.202310344077)
浙江省大学生科技创新活动计划(新苗人才计划)项目(No.2024R410A048).
关键词
浙贝母
干腐病
尖孢镰刀菌
拮抗菌
杀菌剂
Fritillaria thunbergii Miq.
dry rot
Fusarium oxysporum
antagonistic microorganisms
fungicides
