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烟草黑胫病和根黑腐病生防假单胞杆菌的筛选与鉴定
引用本文:千慧敏,文艺,赵辉,倪云霞,刘新涛,邱睿,李小杰,赵新贝,李淑君,刘红彦.烟草黑胫病和根黑腐病生防假单胞杆菌的筛选与鉴定[J].中国生物防治学报,2019,35(6):940-948.
作者姓名:千慧敏  文艺  赵辉  倪云霞  刘新涛  邱睿  李小杰  赵新贝  李淑君  刘红彦
作者单位:1. 河南省农业科学院植物保护研究所, 郑州 450002;2. 烟草行业黄淮烟区烟草病虫害绿色防控重点实验室/河南省农业科学院烟草研究所, 郑州 450002
基金项目:黄淮烟区蚜、烟粉虱和烟草根黑腐病绿色防控关键技术研究与应用(110201502017);河南烟草病虫害绿色防控关键技术研究与应用(201741000027063);黄淮烟区烟草黑胫病高效生防菌剂及抑菌活性物质研究(110201603012)
摘    要:为获得对烟草黑胫病和根黑腐病具有双重防病效果且能够促进烟草生长的假单胞杆菌,采用稀释涂布法从40份土壤样品中分离出201株细菌,通过平板对峙和含毒介质法,筛选出对烟草黑胫病和烟草根黑腐病病原菌均具有良好拮抗作用的菌株PA2101和PG3402。盆栽促生试验表明,菌株PA2101和PG3402能协调地改善烟草地上部分的生长和烟草根系发育,均在一定程度上增加了烟草的株高、叶面积、株鲜重、根鲜重、叶片数和根长,提高了烟草的根冠比和根活力。盆栽试验结果表明,菌株PA2101对烟草黑胫病和根黑腐病的防效分别为70.11%和62.67%,均高于其对照药剂;菌株PG3402对两种病害的防效分别为60.92%和60.00%,与对照药剂相当。抗性标记菌株的定殖试验结果表明,菌株PA2101和PG3402在接种后第29 d能定殖于烟草根际土壤和根内,在烟草茎和叶内也能长时间存在,表明两菌株能够良好定殖。16Sr DNA序列、菌落形态和生理生化性状分析表明,菌株PA2101为铜绿假单胞杆菌Pseudomonas aeruginosa,菌株PG3402为格拉纳达假单胞杆菌Pseudomonas granadensis。综上所述,菌株PA2101和PG3402对烟草具有良好的促生作用,并对烟草黑胫病和根黑腐病有较好的防病效果,是具有生防潜力的菌株。

关 键 词:烟草黑胫病  烟草根黑腐病  生物防治  假单胞杆菌  鉴定
收稿时间:2019-04-02

Screening and Identification of Pseudomonas against Tobacco Black Shank and Tobacco Root Black Rot
QIAN Huimin,WEN Yi,ZHAO Hui,NI Yunxia,LIU Xintao,QIU Rui,LI Xiaojie,ZHAO Xinbei,LI Shujun,LIU Hongyan.Screening and Identification of Pseudomonas against Tobacco Black Shank and Tobacco Root Black Rot[J].Chinese Journal of Biological Control,2019,35(6):940-948.
Authors:QIAN Huimin  WEN Yi  ZHAO Hui  NI Yunxia  LIU Xintao  QIU Rui  LI Xiaojie  ZHAO Xinbei  LI Shujun  LIU Hongyan
Affiliation:1. Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;2. Key Laboratory for Green Preservation&Control of Tobacco Diseases and Pests in Huanghuai Growing Area/Tobacco Research Institute of Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
Abstract:In order to obtain Pseudomonas bacteria with good biocontrol effect on tobacco black shank disease and black root rot disease, which can also promote tobacco growth, 201 biocontrol bacteria were isolated from 40 soil samples. Using plate confrontation method and poison media method, we ultimately obtained bacteria strains PA2101 and PG3402 that had stronger antagonistic effect on the pathogens of tobacco black shank disease and black root rot disease. The pot experiment showed that PA2101 and PG3402 could improve the growth of aboveground plants and root system in a coordinated way. Both of them could increase the plant height, leaf area, aerial fresh weight, root fresh weight, leaf number and root length of tobacco, as well as the root shoot ratio and root activity of tobacco. The control efficacies of PA2101 on tobacco black shank and black root rot were 70.11% and 62.67%, which were better than their fungicide control. And the control efficacies of PG3402 on the two tobacco diseases were 60.92% and 60.00%, which were equivalent to that of fungicide control. Colonization test with resistant label method showed that PA2101 and PG3402 could stably colonize on tobacco rhizosphere soil and root on 29th day after inoculation, and that they could be detected in tobacco stem and leaf, indicating that the two strains could well colonize. On the basis of molecular analysis of 16S rDNA sequences, colonial morphology, as well as physiological and biochemical characteristics, PA2101 and PG3402 were identified as Pseudomonas aeruginosa and Pseudomonas granadensis respectively. PA2101 and PG3402 have good growth-promoting effects and disease prevention effects on tobacco black shank disease and root black rot disease, indicating that they are of great potential in the biocontrol on the two tobacco fungal diseases.
Keywords:tobacco black shank  tobacco black root rot  biocontrol  Pseudomonas sp    identification  
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