一株大蒜根际细菌特性研究及其对田间大蒜产量和土壤酶活性的影响
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摘要
植物根际促生菌(Plant growth-promoting rhizobacteria,PGPR)具有抵抗植物病害,促进植物生长等作用。以前期在山东省金乡县连作大蒜根际土壤中分离获得的根际细菌为研究对象,筛选获得对大蒜根腐病病原菌H5(Setophoma terrestris)和H9 (Fusarium solani)具有较好拮抗效果的菌株DS7,经16S rRNA基因序列分析初步鉴定,该菌株属于芽孢杆菌属(Bacillus)。功能特性研究显示:菌株Bacillus sp. DS7对H5抑制率达到51.72%,对H9抑制率达到42.31%;菌株DS7具有产IAA(19.29±0)μg/mL、产铁载体、水解蛋白、溶磷等能力,但不具有产ACC脱氨酶能力。田间接种试验结果显示:与不接菌的对照相比,接种菌株DS7后大蒜产量提高15.42%;土壤的蔗糖酶活性和脲酶活性高于对照土壤,过氧化氢酶活性低于对照土壤。综上,植物根际促生菌Bacillus sp. DS7具有多种功能特性,可以增加田间大蒜产量,改良土壤,是潜在的微生物肥料生产菌种。
Plant growth-promoting rhizobacteria(PGPR)has the function of resisting plant diseases and promoting plant growth.The rhizosphere bacteria previously isolated from the rhizosphere soil of garlic in continuous cropland in Jinxiang County of Shandong Province were studied.The results showed that the selected strains had good antagonism on the pathogens H5(Setophoma terrestris)and H9(Fusarium solani)of garlic root rot.The strain DS7 was initially identified by the sequence analysis of the 16 S rRNA gene,which belongs to the genus Bacillus.The study of functional properties showed that the inhibition rate of strain Bacillus sp.DS7 inhibition to H5 reached 51.72%,and the inhibition rate to H9 reached 42.31%. The strain DS7 carrys the abilities of producinged IAA(19.29±0)μg/mL,producing iron-producing carrier,hydrolyzing ed protein,and dissolving phosphorus-dissolving ability,but does can't have the ability to produce ACC deaminase. The field inoculation test results showed that the garlic yield increased by 15.42% after inoculation with strain DS7,and the invertase activity and urease activity of soil were higher than that of the control soil,and the catalase activity was lower than that of the control soil.In summary,the plant rhizosphere growth-promoting strain Bacillus sp.DS7 has a variety of functional properties,which can increase the yield of garlic in the field and improve the soil.It is a potential microbe fertilizer production strain.
Plant growth-promoting rhizobacteria(PGPR)has the function of resisting plant diseases and promoting plant growth.The rhizosphere bacteria previously isolated from the rhizosphere soil of garlic in continuous cropland in Jinxiang County of Shandong Province were studied.The results showed that the selected strains had good antagonism on the pathogens H5(Setophoma terrestris)and H9(Fusarium solani)of garlic root rot.The strain DS7 was initially identified by the sequence analysis of the 16 S rRNA gene,which belongs to the genus Bacillus.The study of functional properties showed that the inhibition rate of strain Bacillus sp.DS7 inhibition to H5 reached 51.72%,and the inhibition rate to H9 reached 42.31%. The strain DS7 carrys the abilities of producinged IAA(19.29±0)μg/mL,producing iron-producing carrier,hydrolyzing ed protein,and dissolving phosphorus-dissolving ability,but does can't have the ability to produce ACC deaminase. The field inoculation test results showed that the garlic yield increased by 15.42% after inoculation with strain DS7,and the invertase activity and urease activity of soil were higher than that of the control soil,and the catalase activity was lower than that of the control soil.In summary,the plant rhizosphere growth-promoting strain Bacillus sp.DS7 has a variety of functional properties,which can increase the yield of garlic in the field and improve the soil.It is a potential microbe fertilizer production strain.
引文
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[27]陶光灿,王素英,王玉平,等.芽孢杆菌属(Bacillus sp.)10株细菌混合制剂对4种作物出苗及苗期生长的影响[J].应用与环境生物学报,2003,9(6):598-602.
[28]王娜娜.拮抗性大蒜内生细菌的多样性及其促植物生长特性研究[D].杨凌:西北农林科技大学,2010.
[29]张静,冉晓潇,朱天辉,等.解淀粉芽孢杆菌对山茶灰斑病菌的抑制作用[J].东北林业大学学报,2014,42(7):122-125.
[30]钱一鑫,康冀川,罗乙凯,等.猕猴桃灰霉病拮抗解淀粉芽孢杆菌X17可湿性粉剂的研制[J].中国生物防治学报,2016,32(3):342-348.
[31]吴一晶,林艺芬,林河通,等.生防菌解淀粉芽孢杆菌研究进展[J].包装与食品机械,2012,30(6):49-52.
[32]Gupta G,Panwar J,Jha P N.Natural occurrence of Pseudomonas aeruginosa,a dominant cultivable diazotrophic endophytic bacterium colonizing Pennisetum glaucum.(L.)R.Br.[J].Applied Soil Ecology,2013,64:252-261.
[33]Gao M,Zhou J,Wang E,et al.Multiphasic characterization of a plant growth promoting bacterial strain Burkholderia sp.7016 and its effect on tomato growth in the field[J].Journal of Integrative Agriculture,2015,14(9):1855-1863.
[34]李琬,刘淼,张必弦,等.植物根际促生菌的研究进展及其应用现状[J].中国农学通报,2014,30(24):1-5.
[35]李建宏,李雪萍,张建贵,等.饲用玉米根际促生菌资源筛选及其特性研究[J].草原与草坪,2017,37(1):44-50.
[36]洪常青,何忠俊,鱼海霞.三江并流区暗棕壤酶活性特征研究[J].云南农业大学学报(自然科学版),2013,28(6):857-864.
[37]景宇鹏,李跃进,年佳乐,等.土默川平原不同盐渍化土壤酶活性特征的研究[J].生态环境学报,2013,22(9):1538-1543.
[38]张志丹,赵兰坡.土壤酶在土壤有机培肥研究中的意义[J].土壤通报,2006,(2):2362-2368.
[39]兰宇,韩晓日,战秀梅,等.施用不同有机物料对棕壤酶活性的影响[J].土壤通报,2013,44(1):110-115.
[40]马云华,王秀峰,魏岷,等.黄瓜连作土壤酚类物质积累对土壤微生物和酶活性的影响[J].应用生态学报,2005,16(11):2149-2153.
[41]Woese C R.Bacterial evolution[J].Microbiol Rev,1987,51:221-271.
[42]尹淑丽,张丽萍,习彦花,等.沼渣对土壤微生态结构、土壤酶活及理化性状的影响[J].中国沼气,2017,35(1):72-76.
[43]赵晶,孟庆峰,周连仁,等.长期施用有机肥对草甸碱土土壤酶活性及养分特征的影响[J].中国土壤与肥料,2014,(2):23-26,34.
[44]牛鑫斌,杨慧,孙建光,等.三株固氮类芽孢杆菌的特点及其对中国青菜的产量和土壤酶活性的影响[J/OL].微生物学报,2018,58(7):1213-1223.
[2]Bhattacharyya P N,Jha D K.Plant growth-promoting rhizobacteria(PGPR):emergence in agriculture[J].World Journal of Microbiology and Biotechnology,2012,28(4):1327-1350.
[3]龙伟文,王平,冯新梅.PGPR与AMF相互关系的研究进展[J].应用生态学报,2000,11(2):311-314.
[4]Rana A,Saharan B,Joshi M,et al.Identification of multi-trait PGPR isolates and evaluating their potential as inoculants for wheat[J].Annals of Microbiology,2011,61:893-900.
[5]崔晓双,王伟,张如,等.基于根际营养竞争的植物根际促生菌的筛选及促生效应研究[J].南京农业大学学报,2015,38(6):958-966.
[6]Haiyambo D H,Chimwamurombe P M,Reinhold-Hurek B.Isolation and screening of rhizosphere bacteria from grasses in east Kavango Region of Namibia for plant growth promoting characteristics[J].Current Microbiology,2015,71(5):566-571.
[7]Ahmad F,Ahmad I,Khan M S.Screening of free-living rhizospheric bacteria for their multiple plant growth promoting activities[J].Microbiological Research,2008,163(2):173-181.
[8]Thokchom E,Kalita M C,Talukdar N C.Isolation,screening,characterization,and selection of superior rhizobacterial strains as bioinoculants for seedling emergence and growth promotion of Mandarin orange(Citrus reticulata Blanco)[J].Canadian Journal of Microbiology,2014,60(2):85-92.
[9]Kuklinsky-Sobral J,Araújo W L,Mendes R,et al.Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion[J].Environmental Microbiology,2004,6(12):1244-1251.
[10]Rodríguez H,Fraga R.Phosphate solubilizing bacteria and their role in plant growth promotion[J].Biotechnology Advances,1999,17(4/5):319-339.
[11]贾茹,张迪,马晓东,等.大蒜连作障碍研究进展[J].北方园艺,2014,(19):207-210.
[12]刘素慧,刘世琦,张自坤,等.大蒜连作对其根际土壤微生物和酶活性的影响[J].中国农业科学,2010,43(5):1000-1006.
[13]李宪生,刘留,孙光天.地膜大蒜根腐病的发生与防治[J].现代农业科技,2010,(20):212.
[14]张梦琦,陈云云,张熙,等.多功能植物根际促生菌DD3的功能特性及对大蒜幼苗的促生效果[J].植物营养与肥料学报,2017,23(3):748-756.
[15]孙建光,胡海燕,刘君,等.农田环境中固氮菌的促生潜能与分布特点[J].中国农业科学,2012,45(8):1532-1544.
[16]González-Sánchez M.Evaluation of the effectiveness of biocontrol bacteria against avocado white root rot occurring under commercial greenhouse plant production conditions[J].Biological Control,2013,67:94-100.
[17]孙建光,徐晶,胡海燕,等.中国十三省市土壤中非共生固氮菌微生物菌种资源研究[J].植物营养与肥料学报,2009,15(6):1450-1465.
[18]Amprayn K,Rose M T,Kecskes M,et al.Plant growth promoting characteristics of soil yeast(Candida tropicalis HY)and its effectiveness for promoting rice growth[J].Applied Soil Ecology,2012,61:295-299.
[19]Glick B R.Modulation of plant ethylene levels by the bacterialenzyme ACC deaminase[J].FEMS Microbiology Letter,2005,251(1):1-7.
[20]Schwyn B,Neilands J B.University Chemical assay for the detection and determination of siderophores[J].Analytical Biochemistry,1987,160:47-56.
[21]关松荫.土壤酶及其研究法[M].北京:农业出版社,1986.274-340.
[22]Christopher A D,Soo-Jin K,Soon-Wo K,et al.Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens,Bacillus methylotrophicus,Bacillus amyloliquefaciens subsp.plantarum and'Bacillus oryzicola'are later heterotypic synonyms of Bacillus velezensis based on phylogenomics[J].International Journal of Systematic and Evolutionary Microbiology,2016,66(3):1212-1217.
[23]刘洪波.探讨植物根际促生菌在蔬菜种植中的应用进展[J].种子科技,2017,35(6):44-47.
[24]Ipek M,Pirlak L,Esitken A,et al.plant growth-promoting rhizobacteria(Pgpr)increase yield,growth and nutrition of strawberry under high-calcareous soil conditions[J].Journal of Plant Nutrition,2014,37(7):990-1001.
[25]Xun F,Xie B,Liu S,et al.Effect of plant growth-promoting bacteria(PGPR)and arbuscular mycorrhizal fungi(AMF)inoculation on oats in saline-alkali soil contaminated by petroleum to enhance phytoremediation[J].Environmental Science&Pollution Research International,2014,22(1):598-608.
[26]Kusari S,Singh S,Jayabaskaran C.Biotechnological potential of plant-associated endophytic fungi:hope versus hype[J].Trends in Biotechnology,2014,32(6):297-303.
[27]陶光灿,王素英,王玉平,等.芽孢杆菌属(Bacillus sp.)10株细菌混合制剂对4种作物出苗及苗期生长的影响[J].应用与环境生物学报,2003,9(6):598-602.
[28]王娜娜.拮抗性大蒜内生细菌的多样性及其促植物生长特性研究[D].杨凌:西北农林科技大学,2010.
[29]张静,冉晓潇,朱天辉,等.解淀粉芽孢杆菌对山茶灰斑病菌的抑制作用[J].东北林业大学学报,2014,42(7):122-125.
[30]钱一鑫,康冀川,罗乙凯,等.猕猴桃灰霉病拮抗解淀粉芽孢杆菌X17可湿性粉剂的研制[J].中国生物防治学报,2016,32(3):342-348.
[31]吴一晶,林艺芬,林河通,等.生防菌解淀粉芽孢杆菌研究进展[J].包装与食品机械,2012,30(6):49-52.
[32]Gupta G,Panwar J,Jha P N.Natural occurrence of Pseudomonas aeruginosa,a dominant cultivable diazotrophic endophytic bacterium colonizing Pennisetum glaucum.(L.)R.Br.[J].Applied Soil Ecology,2013,64:252-261.
[33]Gao M,Zhou J,Wang E,et al.Multiphasic characterization of a plant growth promoting bacterial strain Burkholderia sp.7016 and its effect on tomato growth in the field[J].Journal of Integrative Agriculture,2015,14(9):1855-1863.
[34]李琬,刘淼,张必弦,等.植物根际促生菌的研究进展及其应用现状[J].中国农学通报,2014,30(24):1-5.
[35]李建宏,李雪萍,张建贵,等.饲用玉米根际促生菌资源筛选及其特性研究[J].草原与草坪,2017,37(1):44-50.
[36]洪常青,何忠俊,鱼海霞.三江并流区暗棕壤酶活性特征研究[J].云南农业大学学报(自然科学版),2013,28(6):857-864.
[37]景宇鹏,李跃进,年佳乐,等.土默川平原不同盐渍化土壤酶活性特征的研究[J].生态环境学报,2013,22(9):1538-1543.
[38]张志丹,赵兰坡.土壤酶在土壤有机培肥研究中的意义[J].土壤通报,2006,(2):2362-2368.
[39]兰宇,韩晓日,战秀梅,等.施用不同有机物料对棕壤酶活性的影响[J].土壤通报,2013,44(1):110-115.
[40]马云华,王秀峰,魏岷,等.黄瓜连作土壤酚类物质积累对土壤微生物和酶活性的影响[J].应用生态学报,2005,16(11):2149-2153.
[41]Woese C R.Bacterial evolution[J].Microbiol Rev,1987,51:221-271.
[42]尹淑丽,张丽萍,习彦花,等.沼渣对土壤微生态结构、土壤酶活及理化性状的影响[J].中国沼气,2017,35(1):72-76.
[43]赵晶,孟庆峰,周连仁,等.长期施用有机肥对草甸碱土土壤酶活性及养分特征的影响[J].中国土壤与肥料,2014,(2):23-26,34.
[44]牛鑫斌,杨慧,孙建光,等.三株固氮类芽孢杆菌的特点及其对中国青菜的产量和土壤酶活性的影响[J/OL].微生物学报,2018,58(7):1213-1223.