烟草疫霉拮抗放线菌的生防潜力评价  

Evaluation of Biocontrol Potential of the Antagonistic Actinomycetes against Phytophthora parasitica var.nicotianae

作　　者：章舸 彭玉龙 芶剑渝 何楷 李章海[4] 汪章勋[1] 齐永霞[1] ZHANG Ge;PENG Yulong;GOU Jianyu;HE Kai;LI Zhanghai;WANG Zhangxun;QI Yongxia(College Plant Protection,Anhui Agricultural University/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,Anhui 230036,China;Zunyi Branch,Guizhou Tobacco Company,Zunyi 563000,China;Zheng'an County Tobacco Company of Zunyi,Guizhou Province,Zunyi 563000,China;Research Centre of Tobacco and Health,University of Science and Technology of China,Anhui 230052,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,合肥230036 [2]贵州省烟草公司遵义市公司,遵义563000 [3]贵州省遵义市正安县烟草公司,遵义563000 [4]中国科学技术大学烟草与健康研究中心,合肥230052

出　　处：《中国生物防治学报》2023年第3期667-675,共9页Chinese Journal of Biological Control

基　　金：贵州省烟草公司遵义市烟草公司项目复合微生物肥料研制与应用研究(201901)。

摘　　要：由寄生疫霉烟草致病变种Phytophthora parasitica var.nicotianae引起的烟草黑胫病是连作烟田的重要土传病害之一,重发年份常给烟农造成巨大的经济损失。本研究通过稀释分离法从贵州省遵义市烟草黑胫病重病区的健康烟草根际土壤中共分离获得179株放线菌。通过平板对峙培养法筛选得到15株对烟草黑胫病菌菌丝生长有明显抑制作用的菌株。测定了这15个菌株代谢液对烟草疫霉菌丝生长的抑制作用,其中H-3菌株代谢液对烟草疫霉菌丝生长抑制率最高,达到85.68%。研究了H-3代谢液对烟草疫霉游动孢子萌发的抑制作用。结果显示培养6、12和24 h对游动孢子萌发的抑制率分别为72.75%、65.20%和63.45%。结合形态特征及16S rDNA序列分析,初步鉴定H-3菌株属于链霉菌属Streptomyces sp.。盆栽防效和田间防效试验结果表明,施用H-3对烟草黑胫病具有一定的防效,当添加H-3固体发酵物22.5 kg/hm2时田间防效最好,防效可达70.42%。通过荧光定量PCR技术检测了烟草根际土壤中烟草疫霉的量,结果发现,随着H-3用量的增加,根际土壤中烟草疫霉的拷贝数呈下降趋势,说明H-3处理后,能有效降低烟草根际土壤中烟草疫霉的菌量。Tobacco black shank caused by P.parasitica var.nicotianae was one of the important soil-borne diseases in continuous cropping tobacco fields.When the disease occured seriously,it often causes huge economic losses to tobacco farmers.179 strains of actinomycetes were isolated from healthy tobacco rhizosphere soil in tobacco black shank serious disease area in Zunyi City,Guizhou Province.Fifteen strains with obvious inhibitory effects on the mycelial growth of P.parasitica var.nicotianae were screened by dual culture method.The inhibitory effect of the metabolic fluid of these 15 strains on the mycelial growth of P.parasitica var.nicotianae was determined.The results showed that the metabolic fluid of the strain H-3 had the highest inhibitory rate on the mycelial growth,and the inhibition rate was 85.68%.The inhibitory effect of the metabolic fluid strain H-3 on zoospore germination of P.parasitica var.nicotianae was studied.The results showed that the inhibition rates of zoospore germination for 6 h,12 h and 24 h were 72.75%,65.20%and 63.45%,respectively.The strain H-3 was identified as Streptomyces sp.by morphological characteristics and 16S rDNA gene sequence analysis.Pot experiment and field experiment showed that H-3 had a certain control effect on tobacco black shank.The potted control effect was 55.99%.When H-3 solid fermentation was added at a dosage of 22.5 kg/hm2,the field control effect was the best,which could reached 70.42%.The copy numbers of P.parasitica var.nicotianae in tobacco rhizosphere soil was detected by the method of fluorescence quantitative PCR after application of the strain H-3 solid fermentation.The fluorescence quantitative PCR results showed that the copy numbers of P.parasitica var.nicotianae in tobacco rhizosphere soil decreased with the increasing of H-3 dosage,which indicated that the strain H-3 could effectively reduce the number of P.parasitica var.nicotianae in tobacco rhizosphere soil.

关 键 词：烟草黑胫病菌 放线菌 菌丝生长抑制率 拮抗作用 田间防效 荧光定量PCR 

分 类 号：S476[农业科学—农业昆虫与害虫防治]