极端耐盐碱菌株的筛选及其菌肥对盐碱条件下小麦生长和土壤环境的影响
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摘要
为获得具有盐碱地改良应用潜力的耐盐碱菌株,将采集于东营盐碱地的土样稀释涂布于pH 9、盐浓度100 g·L~(-1)的Gibbson改良培养基上,共获得18株细菌.通过提高盐浓度、pH值,最终获得在pH 12、盐浓度20%的条件下仍然可以生长的极端耐盐碱菌株N14.对N14进行形态学、生理生化特征和16S rDNA序列分析鉴定,结果表明:菌株N14为马氏芽孢杆菌.盆栽试验结果表明,与盐碱土(CK)相比,N14菌肥可以显著提高小麦的生物量,株高、鲜重、干重分别提高了21.8%、57.9%、41.7%;显著增加小麦叶绿素a、叶绿素b、叶绿素总量,增长率分别为36.4%、20.0%、31.7%;显著提高盐碱土壤中的蔗糖酶、脲酶和碱性磷酸酶的活性,增长率分别为23.2%、68.8%、106.5%;显著提高小麦根系的超氧化物歧化酶、过氧化物酶和过氧化氢酶的活性,增长率分别为109.6%、17.8%、50%;显著减少小麦根系丙二醛的含量,减少率为39.8%.本研究为极端耐盐碱菌的应用提供了一条新思路,为盐碱地的改良提供了一条新途径.
To obtain salt-alkali tolerant strains which could be potenially used to improve the quality of saline-alkali soil, soil samples collected from Dongying, Shandong Province were diluted and spread to modified Gibbson medium with pH 9 and salt concentration of 100 g·L~(-1). A total of 18 bacteria strains were obtained. By increasing salt concentration and pH, an extremely salt-alkali tole-rant strain N14 was screened which could grow at pH 12 and salt concentration of 20%. We analyzed the morphological, physiological and biochemical characters and 16 S rDNA sequence of N14. The strain N14 was identified as Bacillus marmarensis. N14 bacterial fertilizer significantly increased the biomass of wheat, improved shoot height, fresh weight and dry weight by 21.8%, 57.9% and 41.7%, respectively. The addition of N14 bacterial fertilizer significantly increased the chlorophyll a, chlorophyll b and total chlorophyll in wheat by 36.4%, 20.0% and 31.7%, respectively. It significantly increased the activities of invertase, urease and alkaline phosphatase in saline-alkali soil by 23.2%, 68.8% and 106.5%, respectively. It also significantly increased the activities of superoxide dismutase, peroxidase and catalase in roots by 109.6%, 17.8% and 50%, respectively. The concentration of malondialdehyde in wheat roots was significantly reduced by 39.8%. This study provided an idea for the application of extreme salt-alkali tolerant bacteria and a way for improvement of saline-alkali soil.
To obtain salt-alkali tolerant strains which could be potenially used to improve the quality of saline-alkali soil, soil samples collected from Dongying, Shandong Province were diluted and spread to modified Gibbson medium with pH 9 and salt concentration of 100 g·L~(-1). A total of 18 bacteria strains were obtained. By increasing salt concentration and pH, an extremely salt-alkali tole-rant strain N14 was screened which could grow at pH 12 and salt concentration of 20%. We analyzed the morphological, physiological and biochemical characters and 16 S rDNA sequence of N14. The strain N14 was identified as Bacillus marmarensis. N14 bacterial fertilizer significantly increased the biomass of wheat, improved shoot height, fresh weight and dry weight by 21.8%, 57.9% and 41.7%, respectively. The addition of N14 bacterial fertilizer significantly increased the chlorophyll a, chlorophyll b and total chlorophyll in wheat by 36.4%, 20.0% and 31.7%, respectively. It significantly increased the activities of invertase, urease and alkaline phosphatase in saline-alkali soil by 23.2%, 68.8% and 106.5%, respectively. It also significantly increased the activities of superoxide dismutase, peroxidase and catalase in roots by 109.6%, 17.8% and 50%, respectively. The concentration of malondialdehyde in wheat roots was significantly reduced by 39.8%. This study provided an idea for the application of extreme salt-alkali tolerant bacteria and a way for improvement of saline-alkali soil.
引文
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[14] Mao X (茆璇),Guo J-F (郭江峰).Functional protein research progression in extremophiles.Chinese Bulletin of Life Sciences (生命科学),2018,30(1):107-112 (in Chinese)
[15] Yan H (燕红),Zhong F (钟方),Gao X-L (高新亮),et al.Isolation and screening of saline-alkali tolerant bacterial strains and the biological characteristics and salt-alkali removal efficiency of the strain screened.Chinese Journal of Ecology (生态学杂志),2012,31(4):1000-1008 (in Chinese)
[16] Buchanan RE,Gibbons NE.Bergey’s Manual of Determinative Bacteriology.8th Ed.Beijing:Science Press,1984 (in Chinese)
[17] Zhao L,Xu Y,Lai XH,et al.Screening and characte-rization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters.Brazilian Journal of Micro-biology,2015,46:977-989
[18] Guan S-Y (关松荫).Soil Enzymes and Their Research Methods.Beijing:China Agricultural Press,1986 (in Chinese)
[19] Zheng B-S (郑炳松).Modern Plant Physiology and Biochemistry Research Technology.Beijing:Meteorological Press,2006 (in Chinese)
[20] He J-Z (贺江舟),Zhang L-L (张莉利),Wang J-M (王建明),et al.The preliminary investigation of halophilic and halotolerant bacteria in Tarim Basin.Journal of Tarim University (塔里木大学学报),2007,19(2):53-56 (in Chinese)
[21] Liu M-S (刘敏胜),Xing X-H (邢新会).Transformation methods of alkalophilic Bacillus pseudofirmus.Journal of Chemical Industry and Engineering (化工学报),2006,57(4):922-926 (in Chinese)
[22] Wernick DG,Pontrelli SP,Pollock AW,et al.Sustainable biorefining in wastewater by engineered extreme alkaliphile Bacillus marmarensis.Science Report,2016,6:20224
[23] Luo H (罗欢).Study of Growth Promoting and Salt Tolerance of Bacillus spp.on Plant and the Research of Underlying Mechanisms.Master Thesis.Nanjing:Nanjing Agricultural University,2013 (in Chinese)
[24] Liu L-Y (刘丽英),Liu K-X (刘珂欣),Zhu H (朱浩),et al.Screening of bacteria that antagonize the main pathogen fungi of apple replantation disorders from anaerobic fermented fluid of organic material and its inhibitory effect.Chinese Journal of Applied Ecology (应用生态学报),2018,29(10):3407-3415 (in Chinese)
[25] Wu K,Yuan S,Wang L,et al.Effects of bio-organic fertilizer plus soil amendment on the control of tobacco bacterial wilt and composition of soil bacterial communities.Biology & Fertility of Soils,2014,50:961-971
[26] Chen L (陈琳).Mechanism of Enhanced Plant Salt Tolerance by Bacillus amyloliquefciens SQR9.PhD Thesis.Nanjing:Nanjing Agricultural University,2016 (in Chinese)
[27] Yao XH,MinH,Lü ZH,et al.Influence of acetamiprid on soil enzymatic activities and respiration.European Journal of Soil Biology,2006,42:120-126
[28] Yan L-M (颜路明),Guo X-Q (郭祥泉).Effect of Salinity-alkalinity stress on the rhizosphere soil enzyme activity of camphor seedlings.Soils (土壤),2017,49(4):733-737 (in Chinese)
[29] Habib SH,Kausar H,Saud HM.Plant growth-promoting rhizobacteria enhance salinity stress tolerance in okra through ROS-scavenging enzymes.BioMed Research International,2016,2016:1-10
[30] Zhang A-Q (张爱琴),Pang Q-Y (庞秋颖),Yan X-F (闫秀峰).Advances in salt-tolerance mechanisms of Suaeda plants.Acta Ecologica Sinica (生态学报),2013,33(12):3575-3583 (in Chinese)
[31] Zhang J (张俊),Liu J (刘娟),Zang X-W (臧秀旺).Research of seed germination ability and physiological change of peanut under different storage method.Journal of Agricultural Science and Technology (中国科技导报),2018,20(6):19-27 (in Chinese)
[32] Sreenivasulu N,Grimm B,Wobus U,et al.Differential response of antioxidant compounds to salinity stress in salt-tolerant and salt-sensitive seedlings of foxtail millet (Setaria italica).Physiologia Plantarum,2010,109:435-442
[33] Li Y-M (李玉梅),Guo X-W (郭修武),Dai H-P (代汉萍).Effects of salt-saline stress on contents of osmo-tic adjustment substances and chlorophyll in Rubus crataegifolius seedlings.Nonwood Forest Research (经济林研究),2015,33(1):9-16 (in Chinese)
[34] Ghoulam C,Fares K.Effect of salinity on seed germination and early seedling growth of sugar beet (Beta vulgaris L.).Seed Science & Technology,2001,29:357-364
[2] Sun Q-J (孙芹菊),Ling W (凌玮),Lin S-H (林少华),et al.Studies on the effects of soil properties and vegetable quality after application of biogas slurry.Journal of Nanjing Forestry University (南京林业大学学报),2018,42(5):91-98 (in Chinese)
[3] Xie G-H (谢光辉),Liu Q-Q (刘奇颀),Duan Z-Q (段增强),et al.Review on resource of non-food land suitable for energy plant production in China.Journal of China Agricultural University (中国农业大学学报),2015,20(2):1-10 (in Chinese)
[4] Akhter J,Mahmood K,Malik KA,et al.Amelioration of a saline sodic soil through cultivation of a salt-tolerant grass Leptochloa fusca.Environmental Conservation,2003,30:26-35
[5] Ravindran KC,Venkatesan K,Balakrishnan V,et al.Restoration of saline land by halophytes for Indian soils.Soil Biology & Biochemistry,2007,39:2661-2664
[6] Lu X-Y (路晓筠),Xiang W-D (项卫东),Zheng G-Y (郑光耀),et al.Research progress of saline alkali soil improvement measures.Jiangsu Agricultural Sciences (江苏农业科学),2015,43(12):5-8 (in Chinese)
[7] Song J-Q (宋家清),Zheng X-S (郑秀社),Zhang Q-G (张庆国),et al.Improving effect of microbial orga-nic fertilizer on coastal saline-alkali soil.Northern Horticulture (北方园艺),2010(18):53-55 (in Chinese)
[8] Pang H-C (逄焕成),Li Y-Y (李玉义),Yan H-J (严慧峻),et al.Effects of inoculating different microorga-nism agents on the improvement of salinized soil.Journal of Agro-Environment Science (农业环境科学学报),2009,28(5):951-955 (in Chinese)
[9] Song Y-Z (宋玉珍),An Z-G (安志刚),Zhang Y-H (张玉红),et al.Application of microorganism fertilizer to saline-alkali soil forestation in Daqing City.Journal of Northeast Forestry University (东北林业大学学报),2008,36(7):17-19 (in Chinese)
[10] Li X-L (李晓兰),Wang H-Y (王海鹰),Yang T (杨涛),et al.The function of soil microorganism bacterial fertilizer in the salt and alkaline land on afforest.Journal of Northwest Forestry University (东北林业大学学报).2005,20(4):60-63 (in Chinese)
[11] Santos H,Da CM.Compatible solutes of organisms that live in hot saline environments.Environmental Microbio-logy,2002,4:501-509
[12] Ochsenreiter T,Pfeifer F,Schleper C.Diversity of Archaea in hypersaline environments characterized by molecular-phylogenetic and cultivation studies.Extremophiles,2002,6:267-274
[13] Inipiguri M,Dilibel T,Ajn B,et al.Isolation and phylogenetic analysis of haloalkalophilic bacteria in the black lake of Yuli County in Xinjiang.Microbiology China (微生物学通报),2016,43(12):2601-2608 (in Chinese)
[14] Mao X (茆璇),Guo J-F (郭江峰).Functional protein research progression in extremophiles.Chinese Bulletin of Life Sciences (生命科学),2018,30(1):107-112 (in Chinese)
[15] Yan H (燕红),Zhong F (钟方),Gao X-L (高新亮),et al.Isolation and screening of saline-alkali tolerant bacterial strains and the biological characteristics and salt-alkali removal efficiency of the strain screened.Chinese Journal of Ecology (生态学杂志),2012,31(4):1000-1008 (in Chinese)
[16] Buchanan RE,Gibbons NE.Bergey’s Manual of Determinative Bacteriology.8th Ed.Beijing:Science Press,1984 (in Chinese)
[17] Zhao L,Xu Y,Lai XH,et al.Screening and characte-rization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters.Brazilian Journal of Micro-biology,2015,46:977-989
[18] Guan S-Y (关松荫).Soil Enzymes and Their Research Methods.Beijing:China Agricultural Press,1986 (in Chinese)
[19] Zheng B-S (郑炳松).Modern Plant Physiology and Biochemistry Research Technology.Beijing:Meteorological Press,2006 (in Chinese)
[20] He J-Z (贺江舟),Zhang L-L (张莉利),Wang J-M (王建明),et al.The preliminary investigation of halophilic and halotolerant bacteria in Tarim Basin.Journal of Tarim University (塔里木大学学报),2007,19(2):53-56 (in Chinese)
[21] Liu M-S (刘敏胜),Xing X-H (邢新会).Transformation methods of alkalophilic Bacillus pseudofirmus.Journal of Chemical Industry and Engineering (化工学报),2006,57(4):922-926 (in Chinese)
[22] Wernick DG,Pontrelli SP,Pollock AW,et al.Sustainable biorefining in wastewater by engineered extreme alkaliphile Bacillus marmarensis.Science Report,2016,6:20224
[23] Luo H (罗欢).Study of Growth Promoting and Salt Tolerance of Bacillus spp.on Plant and the Research of Underlying Mechanisms.Master Thesis.Nanjing:Nanjing Agricultural University,2013 (in Chinese)
[24] Liu L-Y (刘丽英),Liu K-X (刘珂欣),Zhu H (朱浩),et al.Screening of bacteria that antagonize the main pathogen fungi of apple replantation disorders from anaerobic fermented fluid of organic material and its inhibitory effect.Chinese Journal of Applied Ecology (应用生态学报),2018,29(10):3407-3415 (in Chinese)
[25] Wu K,Yuan S,Wang L,et al.Effects of bio-organic fertilizer plus soil amendment on the control of tobacco bacterial wilt and composition of soil bacterial communities.Biology & Fertility of Soils,2014,50:961-971
[26] Chen L (陈琳).Mechanism of Enhanced Plant Salt Tolerance by Bacillus amyloliquefciens SQR9.PhD Thesis.Nanjing:Nanjing Agricultural University,2016 (in Chinese)
[27] Yao XH,MinH,Lü ZH,et al.Influence of acetamiprid on soil enzymatic activities and respiration.European Journal of Soil Biology,2006,42:120-126
[28] Yan L-M (颜路明),Guo X-Q (郭祥泉).Effect of Salinity-alkalinity stress on the rhizosphere soil enzyme activity of camphor seedlings.Soils (土壤),2017,49(4):733-737 (in Chinese)
[29] Habib SH,Kausar H,Saud HM.Plant growth-promoting rhizobacteria enhance salinity stress tolerance in okra through ROS-scavenging enzymes.BioMed Research International,2016,2016:1-10
[30] Zhang A-Q (张爱琴),Pang Q-Y (庞秋颖),Yan X-F (闫秀峰).Advances in salt-tolerance mechanisms of Suaeda plants.Acta Ecologica Sinica (生态学报),2013,33(12):3575-3583 (in Chinese)
[31] Zhang J (张俊),Liu J (刘娟),Zang X-W (臧秀旺).Research of seed germination ability and physiological change of peanut under different storage method.Journal of Agricultural Science and Technology (中国科技导报),2018,20(6):19-27 (in Chinese)
[32] Sreenivasulu N,Grimm B,Wobus U,et al.Differential response of antioxidant compounds to salinity stress in salt-tolerant and salt-sensitive seedlings of foxtail millet (Setaria italica).Physiologia Plantarum,2010,109:435-442
[33] Li Y-M (李玉梅),Guo X-W (郭修武),Dai H-P (代汉萍).Effects of salt-saline stress on contents of osmo-tic adjustment substances and chlorophyll in Rubus crataegifolius seedlings.Nonwood Forest Research (经济林研究),2015,33(1):9-16 (in Chinese)
[34] Ghoulam C,Fares K.Effect of salinity on seed germination and early seedling growth of sugar beet (Beta vulgaris L.).Seed Science & Technology,2001,29:357-364