拮抗细菌KRS022的鉴定及对大丽轮枝菌的抑制效果

doi:10.3864/j.issn.0578-1752.2023.04.005

中国农业科学 ›› 2023, Vol. 56 ›› Issue (4): 649-664.doi: 10.3864/j.issn.0578-1752.2023.04.005

拮抗细菌KRS022的鉴定及对大丽轮枝菌的抑制效果

罗万珍¹^,²(), 王丹²^,³(), 齐宏玥²^,⁴, 王彤²^,⁴, 刘政³^,⁵, 田李⁶, 戴小枫²^,³, 陈捷胤²^,³(), 麦合木提江·米吉提¹^,³^,⁷()

¹新疆农业大学农学院/农业农村部西北荒漠绿洲农林外来入侵生物防控重点实验室，乌鲁木齐 830052
²中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室，北京 100193
³中国农业科学院西部农业研究中心，新疆昌吉 831100
⁴牡丹江师范学院生命科学与技术学院，黑龙江牡丹江 157012
⁵石河子大学农学院，新疆石河子 832000
⁶曲阜师范大学生命科学学院，山东曲阜 273165
⁷新疆慧尔农业集团股份有限公司，新疆昌吉 831199

收稿日期:2022-10-02 接受日期:2022-11-16 出版日期:2023-02-16 发布日期:2023-02-24
通信作者: 陈捷胤，E-mail：chenjieyin@caas.cn; 麦合木提江·米吉提，E-mail：287600102@qq.com
联系方式: 罗万珍，E-mail：lwz17690914949@163.com。王丹，E-mail：wangdan_star@163.com。罗万珍和王丹为同等贡献作者。
基金资助:
国家重点研发计划-政府间国际科技创新合作(2022YFE0111300); 国家重点研发计划-政府间国际科技创新合作(2018YFE0112500); 中国博士后科学基金(2020M680783)

Identification of Antagonistic Bacterium Strain KRS022 and Its Inhibition Effect on Verticillium dahliae

LUO WanZhen¹^,²(), WANG Dan²^,³(), QI HongYue²^,⁴, WANG Tong²^,⁴, LIU Zheng³^,⁵, TIAN Li⁶, DAI XiaoFeng²^,³, CHEN JinYin²^,³(), MIJITI Maihemuti¹^,³^,⁷()

¹College of Agronomy, Xinjiang Agricultural University/Key Laboratory of Prevention and Control of Invasive Alien Species in Agriculture & Forestry of the North-western Desert Oasis (Co-construction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Urumqi 830052
²State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193
³Western Agricultural Research Center, Chinese Academy of Agricultural Sciences, Changji 831100, Xinjiang
⁴College of Life Sciences and Technology, Mudanjiang Normal University, Mudanjiang 157012, Heilongjiang
⁵Agricultural College of Shihezhi University, Shihezi 832000, Xinjiang
⁶College of Life Science, Qufu Normal University, Qufu 273165, Shandong
⁷Xinjiang Huier Agriculture Group Co., Ltd., Changji 831199, Xinjiang

Received:2022-10-02 Accepted:2022-11-16 Published:2023-02-16 Online:2023-02-24

摘要/Abstract

摘要：

【目的】明确拮抗细菌KRS022的分类地位及对多种病原真菌的抑制作用，重点研究对作物黄萎病的防治效果及其对病原大丽轮枝菌（Verticillium dahliae）的抑制效果，为抗大丽轮枝菌生防制剂的研发提供资源。【方法】通过形态学、生理生化试验及16S rDNA与gyrB基因序列串联分析确定KRS022的分类地位；采用对峙培养法和对扣熏蒸法测试KRS022对多种真菌的抑制作用；采用发酵菌液平板法、涂布法以及对扣熏蒸法明确KRS022对大丽轮枝菌的平板抑制效果；通过无细胞上清共培养法与对扣熏蒸法明确KRS022对大丽轮枝菌孢子萌发及菌丝形态的影响；通过盆栽试验及大丽轮枝菌生物量测定明确KRS022对棉花和烟草黄萎病的防治效果；利用RT-qPCR法检测KRS022激发植物免疫相关基因的表达特性。【结果】拮抗菌株KRS022为革兰氏阴性细菌，鉴定为产碱假单胞菌（Pseudomonas alcaligenes）。该菌株具有解磷、解钾、固氮功能，可产生嗜铁素，具有蛋白酶、淀粉酶、过氧化氢酶活性，属于好氧产碱型细菌，100 μg·mL^-1氨苄青霉素可作为该菌株的天然抗性筛选条件。KRS022对多种病原真菌（大丽轮枝菌、尖镰孢、禾谷镰孢、稻瘟病菌、灰葡萄孢、胶孢炭疽菌、果生炭疽菌）均具有不同程度的抑菌活性，对峙和对扣培养对大丽轮枝菌Vd991的抑制率分别达75.19%和99.78%。发酵菌液平板法、涂布法以及对扣熏蒸法对大丽轮枝菌的抑制率分别为96.21%、99.72%和99.44%，均能显著抑制Vd991菌丝生长；KRS022无细胞上清液与大丽轮枝菌分生孢子共培养完全抑制孢子萌发，对扣熏蒸后Vd991菌丝发生膨大、变粗。室内盆栽试验结果表明KRS022能够显著抑制棉花和烟草黄萎病的发生，并发现对植株具有促生作用，与清水处理的烟草盆栽相比，施用KRS022菌液的烟草盆栽的叶维度及单叶面积增幅分别达65.7%和146.4%；施用KRS022菌液后大丽轮枝菌在植株中的生物量显著减少，棉花和烟草对照组（单独接种Vd991）中大丽轮枝菌的生物量分别是处理组（KRS022+Vd991）的1.75和2.57倍。同时KRS022能够激发植物防御反应相关基因的表达，经KRS022处理的棉花植株中水杨酸（SA）通路标记基因GhPR1、茉莉酸（JA）通路标记基因GhAOC4和乙烯（ET）通路标记基因GhEIN2均显著上调表达，上调倍数分别是清水处理的7.23、1.69和15.05倍；经KRS022菌液处理后再接种大丽轮枝菌的棉花植株，GhAOC4和GhEIN2被显著诱导表达，上调倍数分别是只接种大丽轮枝菌的68.09和11.87倍；经KRS022无细胞上清液注射处理后NbHSR203、NbHIN1、NbPR1、NbPR2、NbPR5、NbRbohA、NbRbohB上调表达倍数分别是空白LB处理的1.98、2.79、2.52、1.25、1.70、3.28、3.44倍，均显著上调表达。【结论】拮抗细菌KRS022鉴定为产碱假单胞菌，能够产生铁载体，具有解磷、解钾、固氮并促进植物生长的潜力。对多种病原真菌均具有抑制效果，能够抑制大丽轮枝菌菌丝生长及孢子萌发，具有防治黄萎病及激发植物免疫反应的作用，是一株具有开发前景的防治黄萎病等真菌病害的生防微生物资源。

关键词: 产碱假单胞菌, 黄萎病, 大丽轮枝菌, 生物防治, 植物免疫

Abstract:

【Objective】To clarify the taxonomic status of antagonistic bacterium KRS022 and its inhibitory effect on a variety of pathogenic fungi, and to focus on the control efficacy on Verticillium dahliae, and this study will provide important resources for the research and development of biocontrol preparations against V. dahliae. 【Method】The classification of KRS022 was determined by morphological, physiological and biochemical tests, and 16S rDNA-gyrB gene sequence tandem analysis. The inhibitory effects of KRS022 on various fungi were tested by confrontation and fumigation methods. The effect of KRS022 on the morphology of V. dahliae hyphae and conidia was determined by co-culture of cell-free supernatant and fumigation. The control efficacy of KRS022 on Verticillium wilt of cotton and tobacco was determined by pot experiment and biomass measurement. The RT-qPCR method was used to detect the expression levels of plant resistance-related genes. 【Result】The strain KRS022 was gram-negative bacterium, and it was identified as Pseudomonas alcaligenes. KRS022 has the characteristics of phosphorus and potassium solubilization, nitrogen fixation and siderophore production, and the function of protease, amylase and catalase activities. KRS022 belongs to aerobic alkali-producing bacterium, and 100 μg·mL^-1 ampicillin can be used as the screening condition for natural resistance of this strain. KRS022 showed different degrees of antifungal activity against a variety of pathogenic fungi (such as Verticillium dahliae, Fusarium oxysporum, Fusarium graminearum, Magnaporthe oryzae, Botrytis cinerea, Colletotrichum gloeosprioides, Colletotrichum fructicola). The inhibitory rates of confrontation and fumigation against V. dahliae were 75.19% and 99.78%, respectively. The inhibition rates of PDA plates with fermentation liquid, spreading plates and fumigation against Vd991 were 96.21%, 99.72% and 99.44%, respectively. The co-culture of KRS022 cell-free supernatant with conidia of Vd991 completely inhibited spore germination. The hyphae of Vd991 were enlarged and thickened after fumigation treatment of KRS022. The results of pot experiment showed that KRS022 could significantly inhibit the occurrence of Verticillium wilt of cotton and tobacco. After treatment of KRS022 bacterial solution, the biomass of V. dahliae was significantly reduced. The biomass of V. dahliae in the control group (Vd991 alone) was 1.75 and 2.57 folds higher than that in the treatment group (KRS022+Vd991), respectively. Moreover, KRS022 could also promote plants growth, and the leaf dimension and single leaf area of the tobacco treated with KRS022 fermentation broth increased by 65.7% and 146.4%, respectively. At the same time, KRS022 could stimulate the plants resistance-related genes expression. Cotton treated with KRS022 fermentation broth, the salicylic acid (SA) pathway marker gene GhPR1, jasmonic acid (JA) pathway marker gene GhAOC4 and ethylene (ET) pathway marker gene GhEIN2 were significantly up-regulated, the up-regulation folds were 7.23, 1.69 and 15.05 that of water treatment, respectively. Cotton treated with KRS022 fermentation broth and inoculated with V. dahliae, the expression of GhAOC4 and GhEIN2 was significantly induced, and the up-regulation folds were 68.09 and 11.87 that of cotton plants inoculated with V. dahliae, respectively. The up-regulated expression folds of NbHSR203, NbHIN1, NbPR1, NbPR2, NbPR5, NbRbohA and NbRbohB after KRS022 cell-free supernatant injection were 1.98, 2.79, 2.52, 1.25, 1.70, 3.28 and 3.44 that of LB treatment, respectively. 【Conclusion】The antagonistic bacterium KRS022 was identified as P. alcaligenes. It has the characteristics of siderophore production, phosphorus and potassium solubilization, and nitrogen fixation, which suggested KRS022 may have the function of plant growth promotion. It can inhibit the conidia germination and hyphae growth of V. dahliae. In addition, KRS022 can protect plants from V. dahliae infection and trigger plant immunity response. Together, KRS022 is a potential biocontrol microbial resource with development prospect for the prevention and control of Verticillium wilt and other fungal diseases.

Key words: Pseudomonas alcaligenes, Verticillium wilt, Verticillium dahliae, biocontrol, plant immunity

罗万珍, 王丹, 齐宏玥, 王彤, 刘政, 田李, 戴小枫, 陈捷胤, 麦合木提江·米吉提. 拮抗细菌KRS022的鉴定及对大丽轮枝菌的抑制效果[J]. 中国农业科学, 2023, 56(4): 649-664.

LUO WanZhen, WANG Dan, QI HongYue, WANG Tong, LIU Zheng, TIAN Li, DAI XiaoFeng, CHEN JinYin, MIJITI Maihemuti. Identification of Antagonistic Bacterium Strain KRS022 and Its Inhibition Effect on Verticillium dahliae[J]. Scientia Agricultura Sinica, 2023, 56(4): 649-664.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2023.04.005

https://www.chinaagrisci.com/CN/Y2023/V56/I4/649

图/表 10

表1

图1

表2

图2

图3

图4

图5

图6

图7

图8

参考文献 49

[1]	HARTING R, NAGEL A, NESEMANN K, HÖFER A M, BASTAKIS E, KUSCH H, STANLEY C E, STÖCKLI M, KAEVER A, HOFF K J, et al. Pseudomonas strains induce transcriptional and morphological changes and reduce root colonization of Verticillium spp. Frontiers in Microbiology, 2021, 12: 652468. doi: 10.3389/fmicb.2021.652468. doi: 10.3389/fmicb.2021.652468.
[2]	纪晓彬, 王丹, 刘政, 李冉, 宋健, 张丹丹, 陈捷胤, 戴小枫, 林克剑. 芽胞杆菌BvR001对大丽轮枝菌抑制效果研究. 植物保护, 2021, 47(1): 40-47. doi: 10.16688/j.zwbh.2019568. doi: 10.16688/j.zwbh.2019568.
	JI X B, WANG D, LIU Z, LI R, SONG J, ZHANG D D, CHEN J Y, DAI X F, LIN K J. Inhibitory efficacy of Bacillus BvR001 against Verticillium dahliae. Plant Protection, 2021, 47(1): 40-47. doi: 10.16688/j.zwbh.2019568. (in Chinese) doi: 10. 16688/j.zwbh.2019568.
[3]	LI S, XIAO Q, YANG H, HUANG J, LI Y. Characterization of a new Bacillus velezensis as a powerful biocontrol agent against tomato gray mold. Pesticide Biochemistry and Physiology, 2022, 187: 105199. doi: 10.1016/j.pestbp.2022.105199. doi: 10.1016/j.pestbp.2022.105199.
[4]	HASAN N, FARZAND A, HENG Z, KHAN I U, MOOSA A, ZUBAIR M, NA Y, YING S, CANMING T. Antagonistic potential of novel endophytic Bacillus strains and mediation of plant defense against Verticillium wilt in upland cotton. Plants, 2020, 9(11): 1438. doi: 10.3390/plants9111438. doi: 10.3390/plants9111438.
[5]	DEKETELAERE S, TYVAERT L, FRANÇA S C, HÖFTE M. Desirable traits of a good biocontrol agent against Verticillium wilt. Frontiers in Microbiology, 2017, 8: 1186. doi: 10.3389/fmicb.2017.01186. doi: 10.3389/fmicb.2017.01186 pmid: 28729855
[6]	TAO X Y, ZHANG H L, GAO M T, LI M L, ZHAO T, GUAN X Y. Pseudomonas species isolated via high-throughput screening significantly protect cotton plants against Verticillium wilt. AMB Express, 2020, 10(1): 193. doi: 10.1186/s13568-020-01132-1. doi: 10.1186/s13568-020-01132-1.
[7]	PATTEN C L, GLICK B R. Role of Pseudomonas putida indoleacetic acid in development of the host plant root system. Applied and Environmental Microbiology, 2002, 68(8): 3795-3801. doi: 10.1128/ AEM.68.8.3795-3801.2002. doi: 10.1128/ AEM.68.8.3795-3801.2002.
[8]	AFZAL I, SHINWARI Z K, SIKANDAR S, SHAHZAD S. Plant beneficial endophytic bacteria: Mechanisms, diversity, host range and genetic determinants. Microbiological Research, 2019, 221: 36-49. doi: 10.1016/j.micres.2019.02.001. doi: S0944-5013(18)30459-2 pmid: 30825940
[9]	张煜琦, 刘勇, 陈应凤, 曾军, 贺东方, 方守国. 沼泽红假单胞菌PSB-06菌剂对番茄褪绿病的防效及其作用机制. 植物保护学报, 2021, 48(6): 1496-1507. doi: 10.13802/j.cnki.zwbhxb.2021.2021905. doi: 10.13802/j.cnki.zwbhxb.2021.2021905.
	ZHANG Y Q, LIU Y, CHEN Y F, ZENG J, HE D F, FANG S G. Control efficacy and mechanism of fungicide Rhodopseudomonas palustris PSB-06 on tomato chlorosis virus. Journal of Plant Protection, 2021, 48(6): 1496-1507. doi: 10.13802/j.cnki.zwbhxb.2021.2021905. (in Chinese) doi: 10.13802/j.cnki.zwbhxb. 2021.2021905.
[10]	BIESSY A, NOVINSCAK A, ST-ONGE R, LÉGER G, ZBORALSKI A, FILION M. Inhibition of three potato pathogens by phenazine- producing Pseudomonas spp. is associated with multiple biocontrol- related traits. mSphere, 2021, 6(3): e00427-21. doi: 10.1128/mSphere.00427-21. doi: 10.1128/mSphere.00427-21.
[11]	LI C H, SHI L, HAN Q, HU H L, ZHAO M W, TANG C M, LI S P. Biocontrol of Verticillium wilt and colonization of cotton plants by an endophytic bacterial isolate. Journal of Applied Microbiology, 2012, 113(3): 641-651. doi: 10.1111/j.1365-2672.2012.05371.x. doi: 10.1111/j.1365-2672.2012.05371.x pmid: 22726297
[12]	LÓPEZ-ESCUDERO F J, MERCADO-BLANCO J. Verticillium wilt of olive: A case study to implement an integrated strategy to control a soil-borne pathogen. Plant and Soil, 2011, 344: 1-50. doi: 10.1007/s11104-010-0629-2. doi: 10.1007/s11104-010-0629-2.
[13]	BERG G, OPELT K, ZACHOW C, LOTTMANN J, GÖTZ M, COSTA R, SMALLA K. The rhizosphere effect on bacteria antagonistic towards the pathogenic fungus Verticillium differs depending on plant species and site. FEMS Microbiology Ecology, 2006, 56(2): 250-261. doi: 10.1111/j.1574-6941.2005.00025.x. doi: 10.1111/j.1574-6941.2005.00025.x.
[14]	LIU K Z, DING H T, YU Y, CHEN B. A cold-adapted chitinase- producing bacterium from antarctica and its potential in biocontrol of plant pathogenic fungi. Marine Drugs, 2019, 17(12): 695. doi: 10.3390/md17120695. doi: 10.3390/md17120695.
[15]	GÓMEZ-LAMA CABANÁS C, LEGARDA G, RUANO-ROSA D, PIZARRO-TOBÍAS P, VALVERDE-CORREDOR A, NIQUI J L, TRIVIÑO J C, ROCA A, MERCADO-BLANCO J. Indigenous Pseudomonas spp. strains from the olive (Olea europaea L.) rhizosphere as effective biocontrol agents against Verticillium dahliae: From the host roots to the bacterial genomes. Frontiers in Microbiology, 2018, 9: 277. doi: 10.3389/fmicb.2018.00277. doi: 10.3389/fmicb.2018.00277.
[16]	MALDONADO-GONZÁLEZ M M, BAKKER P A, PRIETO P, MERCADO-BLANCO J. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7. Frontiers in Microbiology, 2015, 6: 266. doi: 10.3389/fmicb.2015.00266. doi: 10.3389/fmicb.2015.00266.
[17]	王明元, 徐志周, 刘建福, 李文魁, 林思锻. 伯克霍尔德菌HQB-1抑制香蕉枯萎病菌的活性化合物分离鉴定. 微生物学通报, 2021, 48(6): 1965-1975. doi: 10.13344/j.microbiol.china.200863. doi: 10.13344/j.microbiol.china.200863.
	WANG M Y, XU Z Z, LIU J F, LI W K, LIN S D. Isolation and identification of bioactive compounds of Burkholderia HQB-1 strain inhibiting banana wilt. Microbiology China, 2021, 48(6): 1965-1975. doi: 10.13344/j.microbiol.china.200863. (in Chinese) doi: 10.13344/j.microbiol.china.200863.
[18]	蒋宝贵, 赵斌. 解磷解钾自生固氮菌的分离筛选及鉴定. 华中农业大学学报, 2005, 24(1): 43-48. doi: 10.3321/j.issn:1000-2421.2005.01.011. doi: 10.3321/j.issn:1000-2421.2005.01.011.
	JIANG B G, ZHAO B. Screening and identification of the bacterium which have high efficiency on resolving phosphorus and potassium and in nitrogen fixation. Journal of Huazhong Agricultural University, 2005, 24(1): 43-48. doi: 10.3321/j.issn:1000-2421.2005.01.011. (in Chinese) doi: 10.3321/j.issn:1000-2421.2005.01.011.
[19]	王晓丹, 闵凡祥, 郭梅, 胡林双, 魏琪, 董学志, 李学湛. 马铃薯青枯病菌生化型研究及菌株接种方法的比较. 中国马铃薯, 2010, 24(1): 38-40.
	WANG X D, MIN F X, GUO M, HU L S, WEI Q, DONG X Z, LI X Z. A study on biotypes of Ralstonia solanacearum from potato hosts and comparison of inoculation methods. Chinese Potato Journal, 2010, 24(1): 38-40. (in Chinese)
[20]	东秀珠, 蔡妙英. 常见细菌系统鉴定手册. 北京: 科学出版社, 2001.
	DONG X Z, CAI M Y. Systematic Identification Manual of Common Bacteria. Beijing: Science Press, 2001. (in Chinese)
[21]	BUCHANAN R E, GIBBONS N E. 中国科学院微生物研究所, 译. 8 版. 伯杰细菌鉴定手册. 北京: 科学出版社, 1984.
	BUCHANAN R E, GIBBONS N E. Instituteof Microbiology, Chinese Academy of Sciencestrans. 8th ed. Bergey’s Manual of Determinative Bacteriology. Beijing: Science Press, 1984. (in Chinese)
[22]	王信, 程亮, 王亚艺, 高旭升, 蔡晓剑. 一种由山葵果胶杆菌引起的马铃薯细菌性软腐病. 西南农业学报, 2018, 31(7): 1386-1392. doi: 10.16213/j.cnki.scjas.2018.7.010. doi: 10.16213/j.cnki.scjas.2018.7.010.
	WANG X, CHENG L, WANG Y Y, GAO X S, CAI X J. Bacterial soft rot disease of Solanum tuberosum L. by Pectobacterium wasabiae. Southwest China Journal of Agricultural Sciences, 2018, 31(7): 1386-1392. doi: 10.16213/j.cnki.scjas.2018.7.010. (in Chinese) doi: 10.16213/j.cnki.scjas.2018.7.010.
[23]	李文学, 肖瑞刚, 吕苗苗, 丁宁, 石华荣, 顾沛雯. 葡萄霜霉病菌实时荧光定量PCR检测体系的建立和应用. 中国农业科学, 2019, 52(9): 1529-1540. doi: 10.3864/j.issn.0578-1752.2019.09.005. doi: 10.3864/j.issn.0578-1752.2019.09.005.
	LI W X, XIAO R G, LÜ M M, DING N, SHI H R, GU P W. Establishment and application of real-time PCR for quantitatively detecting Plasmopara viticola in Vitis vinifera. Scientia Agricultura Sinica, 2019, 52(9): 1529-1540. doi: 10.3864/j.issn.0578-1752.2019.09.005. (in Chinese) doi: 10.3864/j.issn.0578-1752.2019. 09.005.
[24]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^{-ΔΔ CT} method. Methods, 2001, 25(4): 402-408. doi: 10.1006/meth.2001.1262. doi: 10.1006/meth.2001.1262.
[25]	王丹. 大丽轮枝菌CFEM类小分子富含半胱氨酸蛋白家族基因功能研究[D]. 北京: 中国农业科学院, 2020.
	WANG D. Functional analysis of CFEM domain-containing small secreted cysteine-rich proteins in Verticillium dahliae[D]. Beijing: Chinese Academy of Agricultural Sciences, 2020. (in Chinese)
[26]	NINH T T, GAO W, TRUSOV Y, ZHAO J R, LONG L, SONG C P, BOTELLA J R. Tomato and cotton G protein beta subunit mutants display constitutive autoimmune responses. Plant Direct, 2021, 5(11): e359. doi: 10.1002/pld3.359. doi: 10.1002/pld3.359 pmid: 34765865
[27]	张丽. 烟草NtabSPL6-2和NtabSPL6-3基因的功能研究[D]. 西安: 西北大学, 2019.
	ZHANG L. Functional research of NtabSPL6-2 and NtabSPL6-3 genes in Nicotiana tabacum[D]. Xi’an: Northwest University, 2019. (in Chinese)
[28]	于世霞. 烟草和番茄中活性氧介导的绒毡层程序性细胞死亡影响花粉的发育[D]. 泰安: 山东农业大学, 2017.
	YU S X. Reactive oxygen species mediated tapetal programmed cell death affects pollen development in tobacco and tomato[D]. Taian: Shandong Agricultural University, 2017. (in Chinese)
[29]	石中全, 杨致邦, 朱黎黎, 马廉举, 蒋仁举, 杨露. 一株地塞米松磷酸钠降解菌的分离鉴定. 中国微生态学杂志, 2014, 26(11): 1250-1256. doi: 10.13381/j.cnki.cjm.201411003. doi: 10.13381/j.cnki.cjm.201411003.
	SHI Z Q, YANG Z B, ZHU L L, MA L J, JIANG R J, YANG L. Isolation and identification of a strain of bacteria degradating dexamethasone sodium phosphate. Chinese Journal of Microecology, 2014, 26(11): 1250-1256. doi: 10.13381/j.cnki.cjm.201411003. (in Chinese) doi: 10.13381/j.cnki.cjm.201411003.
[30]	薛菊兰, 艾彪, 张艳华. 产碱假单胞菌在医院环境中分布情况调查. 中国消毒学杂志, 2011, 28(2): 241.
	XUE J L, AI B, ZHANG Y H. Investigation of the distribution of Pseudomonas alcaligenes in hospital settings. Chinese Journal of Disinfection, 2011, 28(2): 241. (in Chinese)
[31]	华杰, 李艳霞.20株产碱假单胞菌的分离鉴定及药敏分析. 湖北省暨武汉市微生物学会分析微生物专业委员会第十届第五次学术会议论文汇编, 2008: 324-326.
	HUA J, LI Y X.Isolation and identification of 20 strains of Pseudomonas alcaligenes and their susceptibility analysis. Proceedings of the 10th Fifth Academic Conference of Analytical Microbiology Professional Committee of Hubei Province and Wuhan Society of Microbiology, 2008: 324-326. (in Chinese)
[32]	孙崇思, 陈晓敏, 束长龙, 齐放军, 高继国, 张杰. 对大丽轮枝菌具有拮抗作用的萎缩芽胞杆菌的分离和鉴定. 植物保护, 2014, 40(1): 30-37. doi: 10.3969/j.issn.0529-1542.2014.01.005. doi: 10.3969/j.issn.0529-1542.2014.01.005.
	SUN C S, CHEN X M, SHU C L, QI F J, GAO J G, ZHANG J. Isolation and identification of Bacillus atrophaeus antagonistic against Verticillium dahliae. Plant Protection, 2014, 40(1): 30-37. doi: 10.3969/j.issn.0529-1542.2014.01.005. (in Chinese) doi: 10.3969/j.issn.0529-1542.2014.01.005.
[33]	KURANISHI T, SEKIGUCHI J I, YANAGISAWA I, AKIWA M, TOKUNO Y. Development of a new semi-selective lysine-ornithine- mannitol-arginine-charcoal medium for the isolation of Pantoea species from environmental sources in Japan. Microbes and Environments, 2019, 34(2): 136-145. doi: 10.1264/jsme2.ME18128. doi: 10.1264/jsme2.ME18128.
[34]	ZHOU J P, XIE Y Q, LIAO Y H, LI X Y, LI Y M, LI S P, MA X G, LEI S M, LIN F, JIANG W, HE Y Q. Characterization of a Bacillus velezensis strain isolated from Bolbostemmatis Rhizoma displaying strong antagonistic activities against a variety of rice pathogens. Frontiers in Microbiology, 2022, 13: 983781. doi: 10.3389/fmicb.2022.983781. doi: 10.3389/fmicb.2022.983781.
[35]	CESA-LUNA C, BAEZ A, AGUAYO-ACOSTA A, LLANO- VILLARREAL R C, JUÁREZ-GONZÁLEZ V R, GAYTÁN P, BUSTILLOS-CRISTALES M D R, RIVERA-URBALEJO A, MUÑOZ-ROJAS J, QUINTERO-HERNÁNDEZ V. Growth inhibition of pathogenic microorganisms by Pseudomonas protegens EMM-1 and partial characterization of inhibitory substances. PLoS ONE, 2020, 15(10): e0240545. doi: 10.1371/journal.pone.0240545. doi: 10.1371/journal.pone.0240545.
[36]	SURESH P, REKHA M, GOMATHINAYAGAM S, RAMAMOORTHY V, SHARMA M P, SAKTHIVEL P, SEKAR K, VALAN ARASU M, SHANMUGAIAH V. Characterization and assessment of 2, 4- diacetylphloroglucinol (DAPG)-producing Pseudomonas fluorescens VSMKU3054 for the management of tomato bacterial wilt caused by Ralstonia solanacearum. Microorganisms, 2022, 10(8): 1508. doi: 10.3390/microorganisms10081508. doi: 10.3390/microorganisms10081508.
[37]	阳洁, 秦莹溪, 王晓甜, 尹坤, 江院, 袁涛, 谭志远. 广西药用野生稻内生细菌多样性及促生作用. 生态学杂志, 2015, 34(11): 3094-3100. doi: 10.13292/j.1000-4890.20151023.006. doi: 10.13292/j.1000-4890.20151023.006.
	YANG J, QIN Y X, WANG X T, YIN K, JIANG Y, YUAN T, TAN Z Y. Diversity and growth promotion of endophytic bacteria isolated from Orzy officinalis in Guangxi. Chinese Journal of Ecology, 2015, 34(11): 3094-3100. doi: 10.13292/j.1000-4890.20151023.006. (in Chinese) doi: 10.13292/j.1000-4890.20151023.006.
[38]	WANG X, ZHOU X, CAI Z, GUO L, CHEN X, CHEN X, LIU J, FENG M, QIU Y, ZHANG Y, WANG A. A biocontrol strain of Pseudomonas aeruginosa CQ-40 promote growth and control Botrytis cinerea in tomato. Pathogens, 2020, 10(1): 22. doi: 10.3390/pathogens10010022. doi: 10.3390/ pathogens10010022.
[39]	GAO Y, FENG J, WU J, WANG K, WU S, LIU H, JIANG M. Transcriptome analysis of the growth-promoting effect of volatile organic compounds produced by Microbacterium aurantiacum GX14001 on tobacco (Nicotiana benthamiana). BMC Plant Biology, 2022, 22(1): 208. doi: 10.1186/s12870-022-03591-z. doi: 10.1186/s12870-022-03591-z.
[40]	HERNÁNDEZ-LEÓN R, ROJAS-SOLÍS D, CONTRERAS-PÉREZ M, OROZCO-MOSQUEDA M D C, MACÍAS-RODRÍGUEZ L I, REYES-DE LA CRUZ H, VALENCIA-CANTERO E, SANTOYO G. Characterization of the antifungal and plant growth-promoting effects of diffusible and volatile organic compounds produced by Pseudomonas fluorescens strains. Biological Control, 2015, 81: 83-92. doi: 10.1016/j.biocontrol.2014.11.011. doi: 10.1016/j.biocontrol.2014.11.011.
[41]	ZHOU J Y, YUAN J, LI X, NING Y F, DAI C C. Endophytic bacterium-triggered reactive oxygen species directly increase oxygenous sesquiterpenoid content and diversity in Atractylodes lancea. Applied and Environmental Microbiology, 2015, 82(5): 1577-1585. doi: 10.1128/AEM.03434-15. doi: 10.1128/AEM.03434-15.
[42]	杨艺炜, 黎妍妍, 张安盛, 周建云, 李斌, 王静, 陈德鑫. 烟草黑胫病拮抗菌XF10的筛选与鉴定. 烟草科技, 2018, 51(4): 20-27.
	YANG Y W, LI Y Y, ZHANG A S, ZHOU J Y, LI B, WANG J, CHEN D X. Screening and identification of an antagonistic bacterium XF10 against tobacco black shank. Tobacco Science and Technology, 2018, 51(4): 20-27. (in Chinese)
[43]	SAFARA S, HARIGHI B, BAHRAMNEJAD B, AHMADI S. Antibacterial activity of endophytic bacteria against sugar beet root rot agent by volatile organic compound production and induction of systemic resistance. Frontiers in Microbiology, 2022, 13: 921762. doi: 10.3389/fmicb.2022.921762. doi: 10.3389/fmicb.2022.921762.
[44]	BERG G, ROSKOT N, STEIDLE A, EBERL L, ZOCK A, SMALLA K. Plant-dependent genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different Verticillium host plants. Applied and Environmental Microbiology, 2002, 68(7): 3328-3338. doi: 10.1128/AEM.68.7.3328-3338.2002. doi: 10.1128/AEM.68.7.3328-3338.2002.
[45]	赵君洁, 曾卫军, 李艳红, 葛风伟, 杜钰, 袁琳琳, 赵歉歉, 王敏, 谢红桃, 白若翔, 韩生成, 赵和平, 赵惠新. 大丽轮枝菌拮抗芽孢菌株的分离、鉴定及两株菌抑菌特性研究. 北京师范大学学报(自然科学版), 2017, 53(3): 294-300. doi: 10.16360/j.cnki.jbnuns.2017.03.009. doi: 10.16360/j.cnki.jbnuns.2017.03.009.
	ZHAO J J, ZENG W J, LI Y H, GE F W, DU Y, YUAN L L, ZHAO Q Q, WANG M, XIE H T, BAI R X, HAN S C, ZHAO H P, ZHAO H X. Isolation and identification of antagonistic Bacillus spp. against Verticillium dahliae: The antibacterial properties of two strains. Journal of Beijing Normal University (Natural Science Edition), 2017, 53(3): 294-300. doi: 10.16360/j.cnki.jbnuns.2017.03.009. (in Chinese) doi: 10.16360/j.cnki.jbnuns.2017.03.009.
[46]	DUAN Y, CHEN R, ZHANG R, JIANG W, CHEN X, YIN C, MAO Z. Isolation, identification, and antibacterial mechanisms of Bacillus amyloliquefaciens QSB-6 and its effect on plant roots. Frontiers in Microbiology, 2021, 12: 746799. doi: 10.3389/fmicb.2021.746799. doi: 10.3389/fmicb.2021.746799.
[47]	黄艺烁, 谢学文, 石延霞, 柴阿丽, 李磊, 李宝聚. 一株绿针假单胞菌桔黄亚种在防治番茄匍柄霉叶斑病中的应用. 中国生物防治学报, 2021, 37(6): 1265-1275. doi: 10.16409/j.cnki.2095-039x.2021.06.018. doi: 10.16409/j.cnki.2095-039x.2021.06.018.
	HUANG Y S, XIE X W, SHI Y X, CHAI A L, LI L, LI B J. Application of Pseudomonas chlororaphis subsp. aurantiaca against gray leaf spot of tomato. Chinese Journal of Biological Control, 2021, 37(6): 1265-1275. doi: 10.16409/j.cnki.2095-039x.2021.06.018. (in Chinese) doi: 10.16409/j.cnki.2095-039x.2021.06.018.
[48]	沙月霞, 黄泽阳, 马瑞. 嗜碱假单胞菌Ej2对稻瘟病的防治效果及对水稻内源激素的影响. 中国农业科学, 2022, 55(2): 320-328. doi: 10.3864/j.issn.0578-1752.2022.02.007. doi: 10.3864/j.issn.0578-1752.2022.02.007.
	SHA Y X, HUANG Z Y, MA R. Control efficacy of Pseudomonas alcaliphila strain Ej2 against rice blast and its effect on endogenous hormones in rice. Scientia Agricultura Sinica, 2022, 55(2): 320-328. doi: 10.3864/j.issn.0578-1752.2022.02.007. (in Chinese) doi: 10.3864/j.issn.0578-1752.2022.02.007.
[49]	HU L J, WU X Q, WEN T Y, YE J R, QIU Y J, RUI L, ZHANG Y. The key molecular pattern BxCDP1 of Bursaphelenchus xylophilus induces plant immunity and enhances plant defense response via two small peptide regions. Frontiers in Plant Science, 2022, 13: 937473. doi: 10.3389/fpls.2022.937473. doi: 10.3389/fpls.2022.937473.

基因Gene	正向引物 Forward primer (5′-3′)	反向引物 Reverse primer (5′-3′)	参考文献Reference
GhUB7	GAAGGCATTCCACCTGACCAAC	CTTGACCTTCTTCTTCTTGTGCTTG	[26]
GhPR1	ACCTCAACGCTCACAACACA	GGTCCACTGGAGTGCACAAG	[26]
GhEIN2	TTTTGATCTGGTAGCCCCC	CAATATGAAACCTGCCGCAT	[26]
GhAOC4	GGCATCACGGCTGGACTCT	GCGATGGTGGCACTGGC	[26]
NbEF-1α	TGAGTTCGAGGCTGGTATCT	CACTTGGTGGTGTCCATCTT	[25]
NbHSR203	GGCAGTGGAGGAGCTTAAAT	GCTATGTCCCACTCCATTGTTA	[25]
NbHIN1	ATCCTCGGAGTGATTGCATTAG	TGTTGTTTGTGGTGGACAAATC	[25]
NbPR1	CCGCCTTCCCTCAACTCAAC	GCACAACCAAGACGTACTGAG	[25]
NbPR2	AGGTGTTTGCTATGGAATGC	TCTGTACCCACCATCTTGC	[25]
NbPR5	GGGCCAATCTTGGAGCATTA	CAGTCTCCAGTCTCACAATTACC	[27]
NbRbohA	CTTGCTCGTCAACATCGTG	GGAGAAATCTTGTTGAGAGC	[28]
NbRbohB	TAATACAAGGAGGGCATATT	GCAGAACGAGCATCACCT	[28]

特性 Characteristics	KRS022	产碱假单胞菌 P. alcaligenes	特性 Characteristics	KRS022	产碱假单胞菌 P. alcaligenes
铁载体Siderophore	+	+	厌氧试验Anaerobic test	-	-
解磷Phosphorus solubilization	+	+	过氧化氢酶活性 Catalase activity	+	+
解钾Potassium solubilization	+	+	葡萄糖酵解试验 Glycolysis test	-	-
固氮Nitrogen-fixing	+	+	卡那霉素^aKanamycin	-	-
明胶水解Gelatin hydrolysis	+	+	氨苄青霉素^aAmpicillin	+	+
蛋白酶活性Protease activity	+	+	头孢霉素^a Cephalosporin	-	-
淀粉酶活性Amylase activity	+	+	氯霉素^aChloramphenicol	-	-
甲基红试验MR test	-	-	利福平^aRifampicin	-	-
柠檬酸盐利用Citrate utilization	+	+	血平板试验CAMP test	+	+
吲哚试验Indole test	-	-

拮抗细菌KRS022的鉴定及对大丽轮枝菌的抑制效果

Identification of Antagonistic Bacterium Strain KRS022 and Its Inhibition Effect on Verticillium dahliae

RichHTML

PDF

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 10

参考文献 49

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	王炫栋, 宋振, 兰赫婷, 江樱姿, 齐文杰, 刘晓阳, 蒋冬花. 杨梅园土壤优势放线菌的分离及其防病促生功能[J]. 中国农业科学, 2023, 56(2): 275-286.
[2]	沙月霞, 黄泽阳, 马瑞. 嗜碱假单胞菌Ej2对稻瘟病的防治效果及对水稻内源激素的影响[J]. 中国农业科学, 2022, 55(2): 320-328.
[3]	闫多子,蔡霓,王峰,农向群,王广君,涂雄兵,张泽华. 绿僵菌黏附素MAD1体外表达及诱导花生响应的作用[J]. 中国农业科学, 2021, 54(4): 744-753.
[4]	陈洋,赵红怡,闫俊杰,黄剑,高玉林. 马铃薯块茎蛾性信息素化学合成研究现状[J]. 中国农业科学, 2021, 54(3): 556-572.
[5]	曹钰晗,李紫腾,张静怡,张静娜,胡同乐,王树桐,王亚南,曹克强. 我国苹果斑点落叶病菌携带dsRNA分析及一种dsRNA病毒的鉴定[J]. 中国农业科学, 2021, 54(22): 4787-4799.
[6]	赵卫松,郭庆港,苏振贺,王培培,董丽红,胡卿,鹿秀云,张晓云,李社增,马平. 马铃薯健株与黄萎病株根际土壤真菌群落结构及其对碳源利用特征[J]. 中国农业科学, 2021, 54(2): 296-309.
[7]	张小雪,孙天歌,张迎春,陈丽华,张新宇,李艳军,孙杰. 大丽轮枝菌木糖苷酶基因的鉴定及基于HIGS技术的功能分析[J]. 中国农业科学, 2021, 54(15): 3219-3231.
[8]	胡昌雄,范苇,张倩,陈国华,殷红慧,徐天养,杨进波,杨航,吴道慧,张晓明. 基于两性生命表和年龄-阶段捕食率的南方小花蝽对西花蓟马的控制作用[J]. 中国农业科学, 2021, 54(13): 2769-2780.
[9]	李扬凡,邵美琪,刘畅,郭庆港,王培培,陈秀叶,苏振贺,马平. 解淀粉芽孢杆菌HMB33604的抑菌物质及对马铃薯黑痣病的防治效果[J]. 中国农业科学, 2021, 54(12): 2559-2569.
[10]	赵卫松,郭庆港,李社增,王培培,鹿秀云,苏振贺,张晓云,马平. 花铃期棉花黄萎病抗病与感病品种对土壤细菌群落结构的影响[J]. 中国农业科学, 2020, 53(5): 942-954.
[11]	李姝,王杰,黄宁兴,金振宇,王甦,张帆. 捕食性天敌储蓄植物系统研究进展与展望[J]. 中国农业科学, 2020, 53(19): 3975-3987.
[12]	孙琦,何芳,邵胜楠,刘政,黄家风. 棉花黄萎病菌VdHP1的克隆及功能分析[J]. 中国农业科学, 2020, 53(14): 2872-2884.
[13]	张磊,贾琦,巫蔚,赵路评,薛冰,刘欢欢,尚静,雍太文,李庆,杨文钰. 大豆高隆象致病球孢白僵菌菌株BEdy1的鉴定及毒力测定[J]. 中国农业科学, 2020, 53(14): 2974-2982.
[14]	刘海洋, 王伟, 张仁福, 热西达·阿不都热合曼, 姚举. 黄萎病不同发生程度棉田土壤中的真菌群落特征分析[J]. 中国农业科学, 2019, 52(3): 455-465.
[15]	赵卫松,郭庆港,李社增,王亚娇,鹿秀云,王培培,苏振贺,张晓云,马平. 西兰花残体还田对棉花黄萎病防治效果及其对不同生育时期土壤细菌群落的影响[J]. 中国农业科学, 2019, 52(24): 4505-4517.