水稻分蘖期和孕穗期根际反硝化菌群落结构及功能变化
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摘要
通过水稻盆栽试验,分别于水稻分蘖期和孕穗期采集根际与非根际土壤,利用末端限制性片段长度多态技术(T-RFLP)和实时荧光定量PCR(qPCR)技术探究水稻生长对根际反硝化作用和反硝化微生物的影响.结果表明:分蘖期根际土壤的反硝化势显著低于非根际土壤,而孕穗期根际与非根际土壤的反硝化势没有显著性差异.但分蘖期和孕穗期根际土壤中含narG和nosZ基因的微生物数量均显著高于非根际土壤,其中含nosZ基因的反硝化微生物的群落组成结构和多样性对根际环境响应更敏感.说明虽然水稻根系生长会刺激反硝化微生物的生长繁殖,但抑制了根际土壤中一些反硝化微生物的活性,从而降低了根际土壤的反硝化潜势.
We investigated the variation of denitrifying communities in rice rhizosphere at tillering and booting stages in comparison with bulk soils with a pot experiment. The techniques of quantitative polymerase chain reaction(qPCR) and terminal restriction fragment length polymorphism(T-RFLP) were used to measure the abundance and community composition of denitrifiers(narG and nosZ), respectively. The results showed that the potential denitrification activity in the rhizosphere at tillering stage was significantly lower than bulk soils. No significant difference was detected between the rhizosphere and bulk soils at booting stage. The abundance of both narG-and nosZ-containing denitrifying bacteria was significantly higher in rhizosphere than in bulk soils at both tillering and booting stages. In comparison with narG-containing community, community composition and diversity of nosZ-containing bacteria were more sensitive to rice growth. In conclusion, the exudates of rice could induce significantly more denitrifying bacteria in rhizosphere, whose denitrifying activities were related to growth stage of rice. At the period with strong growth, the secretion of roots showed clear restriction to the functions of rhizospheric denitrifiers compared to booting stage.
We investigated the variation of denitrifying communities in rice rhizosphere at tillering and booting stages in comparison with bulk soils with a pot experiment. The techniques of quantitative polymerase chain reaction(qPCR) and terminal restriction fragment length polymorphism(T-RFLP) were used to measure the abundance and community composition of denitrifiers(narG and nosZ), respectively. The results showed that the potential denitrification activity in the rhizosphere at tillering stage was significantly lower than bulk soils. No significant difference was detected between the rhizosphere and bulk soils at booting stage. The abundance of both narG-and nosZ-containing denitrifying bacteria was significantly higher in rhizosphere than in bulk soils at both tillering and booting stages. In comparison with narG-containing community, community composition and diversity of nosZ-containing bacteria were more sensitive to rice growth. In conclusion, the exudates of rice could induce significantly more denitrifying bacteria in rhizosphere, whose denitrifying activities were related to growth stage of rice. At the period with strong growth, the secretion of roots showed clear restriction to the functions of rhizospheric denitrifiers compared to booting stage.
引文
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[3] Yang JX, Yong L, Ye ZH. Root-induced changes of pH, Eh, Fe(Ⅱ) and fractions of Pb and Zn in rhizos-phere soils of four wetland plants with different radial oxygen losses. Pedosphere, 2012, 22: 518-527
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[14] Nicolaisen MH, Risgaard-Petersen N, Revsbech NP, et al. Nitrification-denitrification dynamics and community structure of ammonia oxidizing bacteria in a high yield irrigated Philippine rice field. FEMS Microbiology Ecology, 2004, 49: 359-369
[15] Li Z-G (李振高), Yu S (俞慎), Pan Y-H (潘映华), et al. Horizontal spatial variations of ecological factors of nitrification-denitrification in soil of rice rhizos-phere. Acta Pedologica Sinica (土壤学报), 1999, 36(1): 111-117 (in Chinese)
[16] Kramer SB, Reganold JP, Glover JD, et al. Reduced nitrate leaching and enhanced denitrifier activity and efficiency in organically fertilized soils. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103: 4522-4527
[17] Li Y-L (李奕林), Zhang Y-L (张亚丽), Hu J (胡江), et al. Spatiotemporal variations of nitrification in rhizosphere soil for two different rice cultivars at the seedling stage growing under waterlogged conditions. Acta Ecologica Sinica (生态学报), 2006, 26(5): 1461-1467 (in Chinese)
[18] Kakuda K, Ando H, Konno T. Contribution of nitrogen absorption by rice plants and nitrogen immobilization enhanced by plant growth to the reduction of nitrogen loss through denitrification in rhizosphere soil. Soil Science and Plant Nutrition, 2000, 46: 601-610
[19] Pell M, Stenberg B, Stenstr?m J, et al. Potential denitrification activity assay in soil with or without chloramphenicol? Soil Biology and Biochemistry, 1996, 28: 393-398
[20] Chen Z, Luo XQ, Hu RG, et al. Impact of long-term fertilization on the composition of denitrifier communities based on nitrite reductase analyses in a paddy soil. Microbial Ecology, 2010, 60: 850-861
[21] Chen Z (陈哲). Effect of Long-term Fertilization on Denitrification and Denitrifying Microbial Community. Changsha: Institute of Subtropical Agriculture, Chinese Academy of Sciences, 2010 (in Chinese)
[22] R?sch C, Mergel A, Bothe H. Biodiversity of denitri-fying and dinitrogen-fixing bacteria in an acid forest soil. Applied and Environmental Microbiology, 2002, 68: 3818-3829
[23] Perryman SE, Rees GN, Walsh CJ. Analysis of denitri-fying communities in streams from an urban and non-urban catchment. Aquatic Ecology, 2008, 42: 95-101
[24] Rees GN, Baldwin DS, Watson GO, et al. Ordination and significance testing of microbial community composition derived from terminal restriction fragment length polymorphisms: Application of multivariate statistics. Antonie van Leeuwenhoek, 2004, 86: 339-347
[25] Zhang ZX, Zhang WZ, Yang HC, et al. Elevated N2O emission by the rice roots: Based on the abundances of narG and bacterial amoA genes. Environmental Science and Pollution Research, 2017, 24: 2116-2125
[26] Briones AM, Okabe S, Umemiya Y, et al. Influence of different cultivars on populations of ammonia-oxidizing bacteria in the root environment of rice. Applied and Environmental Microbiology, 2002, 68: 3067-3075
[27] Keeney DR, Sahrawat KL. Nitrogen transformations in flooded rice soils. Nutrient Cycling in Agroecosystems, 1986, 9: 15-38
[28] Henry S, Texier S, Hallet S, et al. Disentangling the rhizosphere effect on nitrate reducers and denitrifiers: Insight into the role of root exudates. Environmental Microbiology, 2008, 10: 3082-3092
[29] Chen HX, Ma GL, Chen ZG, et al. Study on denitrification intensity in rice rhizosphere soil under water management model. Advanced Materials Research, 2014, 1010-1012: 584-587
[30] Chaparro JM, Badri DV, Vivanco JM. Rhizosphere microbiome assemblage is affected by plant development. ISME Journal, 2014, 8: 790-803
[31] Ruiz-Rueda O, Hallin S, Baňeras L. Structure and function of denitrifying and nitrifying bacterial communities in relation to the plant species in a constructed wetland. FEMS Microbiology Ecology, 2009, 67: 308-319
[32] Cavigelli MA, Robertson GP. Role of denitrifier diversity in rates of nitrous oxide consumption in a terrestrial ecosystem. Soil Biology and Biochemistry, 2001, 33: 297-310
[33] Rich JJ, Heichen RS, Bottomley PJ, et al. Community composition and functioning of denitrifying bacteria from adjacent meadow and forest soils. Applied and Environmental Microbiology, 2003, 69: 5974-5982
[34] Li H, Su JQ, Yang XR, et al. Distinct rhizosphere effect on active and total bacterial communities in paddy soils. Science of the Total Environment, 2019, 649: 422-430
[35] Bertin C, Yang XH, Weston LA. The role of root exudates and allelochemicals in the rhizosphere. Plant and Soil, 2003, 256: 67-83
[36] Henry S, Texier S, Hallet S, et al. Disentangling the rhizosphere effect on nitrate reducers and denitrifiers: Insight into the role of root exudates. Environmental Microbiology, 2008, 10: 3082-3092
[37] Lynch JM, Whipps JM. Substrate flow in the rhizos-phere. Plant and Soil, 1990, 129: 1-10
[2] Lu H-Y (陆海雅), Zhang F-S (张福锁). The advances in rhizosphere microbiology. Acta Pedologica Sinica (土壤学报), 2006, 38(2): 113-121 (in Chinese)
[3] Yang JX, Yong L, Ye ZH. Root-induced changes of pH, Eh, Fe(Ⅱ) and fractions of Pb and Zn in rhizos-phere soils of four wetland plants with different radial oxygen losses. Pedosphere, 2012, 22: 518-527
[4] Bru D, Ramette A, Saby N, et al. Determinants of the distribution of nitrogen-cycling microbial communities at the landscape-scale. ISME Journal, 2011, 5: 532-542
[5] Carminati A, Moradi AB, Vetterlein D, et al. Dynamics of soil water content in the rhizosphere. Plant and Soil, 2010, 332: 163-176
[6] Bird JA, Herman DJ, Firestone MK. Rhizosphere pri-ming of soil organic matter by bacterial groups in a grassland soil. Soil Biology and Biochemistry, 2011, 43: 718-725
[7] Luo Y-Q (罗永清), Zhao X-Y (赵学勇), Li M-X (李美霞). Ecological effect of plant root exudates and related affecting factors: A review. Chinese Journal of Applied Ecology (应用生态学报), 2012, 23(12): 3496-3504 (in Chinese)
[8] Jones DL. Rhizosphere Congress 2004: Perspectives and challenges, Munich, September 2004. Soil Biology and Biochemistry, 2006, 38: 1177
[9] Philippot L, Hallin S, B?rjesson G, et al. Biochemical cycling in the rhizosphere having an impact on global change. Plant and Soil, 2009, 321: 61-81
[10] Yang H, Sheng R, Zhang Z, et al. Responses of nitri-fying and denitrifying bacteria to flooding-drying cycles in flooded rice soil. Applied Soil Ecology, 2016, 103: 101-109
[11] Liu JB, Hou HJ, Sheng R, et al. Denitrifying communities differentially respond to flooding drying cycles in paddy soils. Applied Soil Ecology, 2012, 62: 155-162
[12] Zou JW, Huang Y, Zheng XH, et al. Quantifying direct N2O emissions in paddy fields during rice growing season in mainland China: Dependence on water regime. Atmospheric Environment, 2007, 41: 8030-8042
[13] Voesenek LACJ, Colmer TD, Pierik R, et al. How plants cope with complete submergence. New Phytologist, 2006, 170: 213-226
[14] Nicolaisen MH, Risgaard-Petersen N, Revsbech NP, et al. Nitrification-denitrification dynamics and community structure of ammonia oxidizing bacteria in a high yield irrigated Philippine rice field. FEMS Microbiology Ecology, 2004, 49: 359-369
[15] Li Z-G (李振高), Yu S (俞慎), Pan Y-H (潘映华), et al. Horizontal spatial variations of ecological factors of nitrification-denitrification in soil of rice rhizos-phere. Acta Pedologica Sinica (土壤学报), 1999, 36(1): 111-117 (in Chinese)
[16] Kramer SB, Reganold JP, Glover JD, et al. Reduced nitrate leaching and enhanced denitrifier activity and efficiency in organically fertilized soils. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103: 4522-4527
[17] Li Y-L (李奕林), Zhang Y-L (张亚丽), Hu J (胡江), et al. Spatiotemporal variations of nitrification in rhizosphere soil for two different rice cultivars at the seedling stage growing under waterlogged conditions. Acta Ecologica Sinica (生态学报), 2006, 26(5): 1461-1467 (in Chinese)
[18] Kakuda K, Ando H, Konno T. Contribution of nitrogen absorption by rice plants and nitrogen immobilization enhanced by plant growth to the reduction of nitrogen loss through denitrification in rhizosphere soil. Soil Science and Plant Nutrition, 2000, 46: 601-610
[19] Pell M, Stenberg B, Stenstr?m J, et al. Potential denitrification activity assay in soil with or without chloramphenicol? Soil Biology and Biochemistry, 1996, 28: 393-398
[20] Chen Z, Luo XQ, Hu RG, et al. Impact of long-term fertilization on the composition of denitrifier communities based on nitrite reductase analyses in a paddy soil. Microbial Ecology, 2010, 60: 850-861
[21] Chen Z (陈哲). Effect of Long-term Fertilization on Denitrification and Denitrifying Microbial Community. Changsha: Institute of Subtropical Agriculture, Chinese Academy of Sciences, 2010 (in Chinese)
[22] R?sch C, Mergel A, Bothe H. Biodiversity of denitri-fying and dinitrogen-fixing bacteria in an acid forest soil. Applied and Environmental Microbiology, 2002, 68: 3818-3829
[23] Perryman SE, Rees GN, Walsh CJ. Analysis of denitri-fying communities in streams from an urban and non-urban catchment. Aquatic Ecology, 2008, 42: 95-101
[24] Rees GN, Baldwin DS, Watson GO, et al. Ordination and significance testing of microbial community composition derived from terminal restriction fragment length polymorphisms: Application of multivariate statistics. Antonie van Leeuwenhoek, 2004, 86: 339-347
[25] Zhang ZX, Zhang WZ, Yang HC, et al. Elevated N2O emission by the rice roots: Based on the abundances of narG and bacterial amoA genes. Environmental Science and Pollution Research, 2017, 24: 2116-2125
[26] Briones AM, Okabe S, Umemiya Y, et al. Influence of different cultivars on populations of ammonia-oxidizing bacteria in the root environment of rice. Applied and Environmental Microbiology, 2002, 68: 3067-3075
[27] Keeney DR, Sahrawat KL. Nitrogen transformations in flooded rice soils. Nutrient Cycling in Agroecosystems, 1986, 9: 15-38
[28] Henry S, Texier S, Hallet S, et al. Disentangling the rhizosphere effect on nitrate reducers and denitrifiers: Insight into the role of root exudates. Environmental Microbiology, 2008, 10: 3082-3092
[29] Chen HX, Ma GL, Chen ZG, et al. Study on denitrification intensity in rice rhizosphere soil under water management model. Advanced Materials Research, 2014, 1010-1012: 584-587
[30] Chaparro JM, Badri DV, Vivanco JM. Rhizosphere microbiome assemblage is affected by plant development. ISME Journal, 2014, 8: 790-803
[31] Ruiz-Rueda O, Hallin S, Baňeras L. Structure and function of denitrifying and nitrifying bacterial communities in relation to the plant species in a constructed wetland. FEMS Microbiology Ecology, 2009, 67: 308-319
[32] Cavigelli MA, Robertson GP. Role of denitrifier diversity in rates of nitrous oxide consumption in a terrestrial ecosystem. Soil Biology and Biochemistry, 2001, 33: 297-310
[33] Rich JJ, Heichen RS, Bottomley PJ, et al. Community composition and functioning of denitrifying bacteria from adjacent meadow and forest soils. Applied and Environmental Microbiology, 2003, 69: 5974-5982
[34] Li H, Su JQ, Yang XR, et al. Distinct rhizosphere effect on active and total bacterial communities in paddy soils. Science of the Total Environment, 2019, 649: 422-430
[35] Bertin C, Yang XH, Weston LA. The role of root exudates and allelochemicals in the rhizosphere. Plant and Soil, 2003, 256: 67-83
[36] Henry S, Texier S, Hallet S, et al. Disentangling the rhizosphere effect on nitrate reducers and denitrifiers: Insight into the role of root exudates. Environmental Microbiology, 2008, 10: 3082-3092
[37] Lynch JM, Whipps JM. Substrate flow in the rhizos-phere. Plant and Soil, 1990, 129: 1-10
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