减量施氮对玉米/大豆套作系统中作物氮素吸收及土壤氨氧化与反硝化细菌多样性的影响

doi:10.3864/j.issn.0578-1752.2017.06.009

摘要/Abstract

摘要： 【目的】揭示玉米/大豆套作系统下作物根际土壤细菌数量及群落多样性变化特征与土壤总氮含量、作物氮素吸收之间的关系，为禾/豆间（套）作减肥增效生产提供理论和技术支撑。【方法】大田试验于2013—2015年进行，采用两因素裂区设计，主因素为种植模式，设玉米单作（MM）、大豆单作（SS）和玉米/大豆套作（IMS）；副因素为玉米、大豆施氮总量，设不施氮（NN：0）、减量施氮（RN：180 kg·hm^-2）和常量施氮（CN：240 kg·hm^-2）。在玉米V12期、VT期和R6期，大豆V5期、R2期、R5期和R8期，利用稀释平板法和凯氏定氮法测定各作物根际土壤细菌数量、非根际土壤和植株总氮含量；结合克隆文库和荧光定量PCR技术研究各处理氨氧化细菌（amoA基因）、反硝化细菌（nirS基因）多样性及其基因丰度。【结果】与相应的单作相比，套作玉米（IM）的作物根际土壤细菌数量提高2.6%，套作大豆提高12.9%；套作玉米土壤总氮含量和植株吸氮量分别提高13.39%和2.10%，大豆的分别降低5.81%和3.24%；套作玉米、大豆的amoA基因丰度比单作增加了38.5%、64.8%，nirS基因丰度比单作提高57.77%、126.39%。各施氮水平间，RN的玉米根际土壤细菌数量比NN和CN的分别提高9.6%和9.8%，大豆的分别提高11.7%和11.0%；施氮提高了玉米、大豆植株吸氮量和土壤总氮含量，单作玉米随施氮量的增加而增加，套作玉米及单、套作大豆的均在RN下最高；减量施氮提高了玉米、大豆amoA基因多样性指数和单作玉米nirS基因多样性指数，降低了套作玉米和单套作大豆nirS基因多样性指数。【结论】减量施氮有利于增加玉米/大豆套作系统中作物根际土壤细菌数量，调节氨氧化细菌和反硝化细菌群落结构及多样性，改善土壤氮素转化过程，促进玉米、大豆对氮素的吸收，实现节肥增效。

关键词: 玉米/大豆套作, 减量施氮, 细菌多样性, 氮素吸收

Abstract: 【Objective】The aim of this study is to reveal the variation characteristics of soil bacteria quantity and community in crop rhizosphere, and the relationship among bacteria quantity and community, soil total N content, crops N uptake in maize/soybean relay strip intercropping system, which will provide theoretical and technology support to reduce fertilizer input and efficient production in cereal/legumes intercropping system.【Method】Field experiments were conducted with two factors split-plot designed from 2013 to 2015, the main factor was planting patterns which included monoculture maize (MM), solo soybean (SS) and maize/soybean relay strip intercropping system (IMS), and the sub-factor total N for maize and soybean were no nitrogen 0 (NN), reduced nitrogen 180 kg N·hm^-2 (RN), conventional nitrogen 240 kg N·hm^-2 (CN), respectively. At V12, VT, R6 stages of maize and V5, R2, R5, R8 stages of soybean, the rhizosphere bacteria quantity was investigated by dilution-plate method, non-rhizosphere soil and crops shoot total N contents were determined by Kjeldahl method, and the diversity and gene abundance of ammonia oxdizing bacteria (amoA) and denitrifying bacteria (nirS) under different N fertilization rates were measured by combining clone library with FQ-PCR. 【Result】 The bacteria quantity in the rhizosphere of intercropped maize (IM) was 2.6% higher, and that of intercropped soybean (IS) was 12.9% higher compared with the corresponding monoculture. The total soil and shoot N contents of intercropped maize were increased by 13.39% and 2.10%, and that of intercropped soybean were decreased by 5.81% and 3.24% in contrast with corresponding monoculture, respectively. The gene abundance of ammonia oxdizing bacteria (amoA) of intercropped maize and soybean was 38.5% and 64.8% higher, and the gene abundance of denitrifying bacteria (nirS) of intercropped maize and soybean was 57.77% and 126.39% higher in contrast with corresponding monoculture, respectively. Under different N fertilization rates, the bacteria quantity in the rhizosphere of maize was 9.6% and 9.8% higher, and that of soybean was 11.7% and 11.0% greater in RN compared NN with CN, respectively. In addition, reduced N fertilization increased the diversity index of amoA gene in the rhizosphere of maize and soybean, and increased the diversity index of nirS of monoculture maize, moreover, reduced N fertilization decreased the diversity index of nirS of intercropped maize and soybean. 【Conclusion】 Reduced N fertilization contributed to increase bacteria quantity in the rhizosphere under maize/soybean relay strip intercropping system, improved the progress of soil N transfer, promoted the N uptake of maize and soybean thereby achieved saving fertilizer.

Key words: maize-soybean relay strip intercropping system, reduced N, bacterial diversity, N uptake

周丽，付智丹，杜青，陈平，杨文钰，雍太文. 减量施氮对玉米/大豆套作系统中作物氮素吸收及土壤氨氧化与反硝化细菌多样性的影响[J]. 中国农业科学, 2017, 50(6): 1076-1087.

ZHOU Li, FU ZhiDan, DU Qing, CHEN Ping, YANG WenYu, YONG TaiWen. Effects of Reduced N Fertilization on Crop N Uptake, Soil Ammonia Oxidation and Denitrification Bacteria Diversity in Maize/Soybean Relay Strip Intercropping System[J]. Scientia Agricultura Sinica, 2017, 50(6): 1076-1087.

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.0578-1752.2017.06.009

https://www.chinaagrisci.com/CN/Y2017/V50/I6/1076

参考文献

[1] 陈文新. 土壤和环境微生物学. 北京: 北京农业大学出版社,1990.

CHEN W X. Soil and Environmental Microbiology. Beijing: Beijing Agricultural University Press, 1990. (in Chinese)

[2] 徐建文, 左淑珍. 保护性耕作对旱作玉米土壤微生物和酶活性影响的探讨. 黑龙江科技信息, 2009(8): 113.

XU J W, ZUO S Z. Effects of conservation tillage on soil microorganism and enzyme activity of dry-land corn. Heilongjiang Science and Technology Information, 2009(8): 113. (in Chinese)

[3] 周修冲, 刘国坚, Portch S. 平衡施肥在广东“三高”农业中的作用. 广东农业科学, 1998(1): 32-34.

ZHOU X C, LIU G J, PORTCH S. Effects of balance fertilization on Guangdong three-high agriculture. Guangdong Agricultural Science, 1998(1): 32-34. (in Chinese)

[4] 雍太文, 杨文钰, 向达兵, 陈小容. 不同种植模式对作物根系生长、产量及根际土壤微生物数量的影响. 应用生态学报, 2012, 23(1): 125-132.

YONG T W, YANG W Y, XIANG D B, CHEN X R. Effects of different cropping modes on crop root growth, yield, and rhizosphere soil microbes’ number. Chinese Journal of Applied Ecology, 2012, 23(1): 125-132. (in Chinese)

[5] 张向前, 黄国勤, 卞新民, 赵其国. 施氮和隔根对玉米植株生长、产量和根际微生物的影响. 应用生态学报, 2012, 23(12): 3369-3376.

ZHANG X Q, HUANG G Q, BIAN X M, ZHAO G Q. Effects of nitrogen fertilization and root separation on the plant growth and grain yield of maize and its rhizosphere microorganisms. Chinese Journal of Applied Ecology, 2012, 23(12): 3369-3376. (in Chinese)

[6] CRECCHIO C, CURCI M, PELLEGRINO A. Soil microbial dynamics and genetic diversity in soil under monoculture wheat grown in different long-?term management systems. Soil Biology and Biochemistry, 2007, 39(6): 1391-1400.

[7] 刘振香, 刘鹏, 贾绪存, 程乙, 董树亭, 赵斌, 张吉旺, 杨今胜. 不同水肥处理对夏玉米田土壤微生物特性的影响. 应用生态学报, 2015, 26(1): 113-121.

LIU Z X, LIU P, JIA X C, CHENG Y, DONG S T, ZHAO B, ZHANG J W, YANG L S. Effects of irrigation and fertilization on soil microbial properties in summer maize field. Chinese Journal of Applied Ecology, 2015, 26(1): 113-121. (in Chinese)

[8] 涂勇, 杨文钰, 刘卫国, 雍太文, 江连强, 王小春. 大豆与烤烟不同套作年限对根际土壤微生物数量的影响. 作物学报, 2015, 41(5): 733-742.

TU Y, YANG W Y, LIU W G, YONG T W, JIANG L Q, WANG X C. Effects of relay strip intercropping years between flue-cured tobacco and soybean on rhizospheric microbes quanties. Acta Agronomica Sinica, 2015, 41(5): 733-742. (in Chinese)

[9] Mei P P, Gui L G, Wang P, LI L. Maize/faba bean intercropping with rhizobia inoculation enhances productivity and recovery of fertilizer P in a reclaimed desert soil. Field Crops Research,2012, 130(11): 19-27.

[10] 王俊华, 尹睿, 张华勇, 林先贵, 陈瑞蕊, 钦绳武. 长期定位施肥对农田土壤酶活性及相关因素的影响. 生态环境, 2007, 26(6): 922-925.

WANG J H , YIN R, ZHENG H Y, LIN X G, CHEN R R, QIN S W. Changes in soil enzyme activities, microbial biomass, and soil nutrition status in response to fertilization regimes in a long-term field experiments. Ecological Environment, 2001, 26(6): 922-925. (in Chinese)

[11] 孟庆英, 于忠和, 贾会彬, 张春峰, 朱宝国, 王囡囡, 高雪冬. 控释肥对大豆根际土壤微生物数目、肥力及大豆农艺性状的影响. 大豆科学, 2011, 30(5): 827-829.

Meng Q Y, Yu Z H, Jia H B, ZHANG C F, ZHU B G, WANG N N, GAO X D. Effects of controlled? release fertilizer on rhizosphere soil microorganisms, soil fertility and agronomic characters of soybean. Soybean Science, 2011, 30(5): 827-829.(in Chinese)

[12] 辜运富, 云翔, 张小平, 涂仕华, 孙锡发, Kristina Lindstrom. 不同施肥处理对石灰性紫色土微生物数量及氨氧化细菌群落结构的影响. 中国农业科学, 2008, 41(12): 4119-4126.

GU Y F, YUN X, ZHANG X P, TU S H, SUN X F, KRISTINA LINDSTROM. Effect of different fertilizer treatments on soil microbes and ammonium oxidizing bacteral community in a calcareous purple paddy soil. Scientia Agricultura Sinica, 2008, 41(12): 4119-4126. (in Chinese)

[13] 曾希柏, 王亚男, 王玉忠, 白玲玉, 李莲芳, 段然, 苏世鸣, 吴翠霞. 施肥对设施菜地nirK型反硝化细菌群落结构和丰度的影响. 应用生态学报, 2014, 25(2): 505-514.

ZENG X B, WANG Y N, WANG Y Z, BAI L Y, LI L F, DUAN R, SU S M, WU C X. Effects of different fertilization regimes on abundance and community structure of the nirk-type denitrifying bacteria in greenhouse vegetable soils. Chinese Journal of Applied Ecology,2014, 25(2): 505-514. (in Chinese)

[14] 郭九信, 杨瑛, 周毅, 段立珍, 汪建飞. 铵硝态氮单独供应对苗期淹水胁迫玉米氮磷钾累积的影响. 西北农林科技大学学报(自然科学版), 2012, 40(8): 185-192.

GUO J X, YANG Y, ZHOU Y, DUAN L Z, WANG J F. Effect of ammonium and nitrate sole supply and nitrogen levels on nitrogen, phosphorus and potassium content and accumulation of maize(Zea mays L) under water-logging stress and seeding stage. Journal of Northwest A ＆ F University(Natural Science Edition), 2012, 40(8): 185-192. (in Chinese)

[15] 汪峰, 曲浩丽, 丁玉芳, 孙波, 崔中利, 曹慧. 三种农田土壤中氨氧化细菌aomA基因多样性比较分析. 土壤学报, 2012, 49(2): 347-352.

WANG F, QU H L, DING Y F, SUN B, CUI Z L, CAO H. Comparative analysis of ammonia monooxygenase (amoA) gene diversity in three cultivated soils. Acta Pedologica Sinica, 2012, 49(2): 347-352. (in Chinese)

[16] Norton J M, Alzerreca J J, Suwa Y C. Diversity ofammonia monooxygenase operon in autotrophic ammonia-oxidizing bacteria. Archives of Microbiology, 2002, 177(2): 139-149.

[17] 尹昌, 范分良, 李兆君. 长期施用有机肥和无机肥对黑土nirS型反硝化菌种群结构和丰度的影响. 环境科学, 2012, 33(11): 3967-3974.

YIN C, FAN F L, LI Z J. Influences of long-term application of organic and inorganic fertilizers on the composition and abundance of nirS-type denitrifiers in black soil. Environmental Science, 2012, 33(11): 3967-3974. (in Chinese)

[18] YANG F, HUANG S, GAO R C, LIU W G, YONG T W, WU X L, YANG W Y. Growth of soybean seedlings in relay strip intercropping systems in relation to light quantity and red: far-red ratio. Field Crops Research, 2014, 155: 245-253.

[19] NGWIRA A R, AUNE J B, MKWINDA S. On farm evaluation of yield and economic benefit of short term maize legume intercropping systems under conservation agriculture in Malawi. Field Crops Research, 2012, 132(3): 149-157.

[20] MAO L L, ZHANG L Z, LI W Q. Yield advantage and water saving in maize/pea intercrop. Field Crops Research, 2012, 138(3): 11-20.

[21] YONG T W, LIU X M, YANG F. Characteristics of nitrogen uptake, use and transfer in a wheat-maize-soybean relay intercropping system. Plant Production Science, 2015, 18(3): 388-397.

[22] BETENCOURT E, DUPUTEL M, COLOMB B. Intercropping promotes the ability of durum wheat and chick pea to increase rhizosphere phosphorus availability in a low P soil. Soil Biology & Biochemistry, 2012, 46(1): 181-190.

[23] LITHOURGIDIS A S, VLACHOSTERGIOS D N, DORDASC C A. Dry matter yield, nitrogen content, and competition in pea-cereal intercropping systems. European Journal of Agronomy, 2011, 34(4): 287-294.

[24] 刘小明, 雍太文, 刘文钰, 苏本营, 宋春, 杨峰, 王小春, 杨文钰. 减量施氮对玉米/大豆套作体系土壤氮素残留和氮肥损失的影响. 应用生态学报, 2014, 25(8): 2267-2274.

LIU X M, YONG T W, LIU W Y, SU B Y, SONG C, YANG F ,WANG X C, YANG W Y. Effects of reduced N application on soil N residue and N loss in maize-soybean relay strip intercropping system. Chinese Journal of Applied Ecology, 2014, 25(8): 2267-2274. (in Chinese)

[25] 苏本营, 陈圣宾, 李永庚, 杨文钰. 间套作种植提升农田生态系统服务功能. 生态学报, 2013, 33(14): 4505-4514.

SU B Y, CHEN S B, LI Y G, YANG W Y. Intercropping enhances the farmland ecosystem services. Acta Ecologica Sinica, 2013, 33(14): 4505-4514. (in Chinese)

[26] 雍太文, 刘小明, 宋春, 周丽, 李星辰. 种植方式对玉米-大豆套作体系中作物产量、养分吸收和种间竞争的影响. 中国生态农业学报, 2015, 23(6): 659-667.

YONG T W, LIU X M, SONG C, ZHOU L, LI X C. Effect of planting patterns on crop yield, nutrients uptake and interspecific competition in maize-soybean relay strip intercropping system. Chinese Journal of Eco-Agriculture, 2015, 23(6): 659-667. (in Chinese)

[27] 雍太文, 刘小明, 刘文钰, 周丽, 宋春, 杨峰, 蒋利, 王小春, 杨文钰. 减量施氮对玉米-大豆套作系统下作物氮素吸收和利用效率的影响. 生态学报, 2015, 35(13): 4473-4482.

YONG T W, LIU X M, LIU W Y, ZHOU L, SONG C, YANG F, JIANG L, WANG X C, YANG W Y. Effects of reduced nitrogen application on nitrogen uptake and utilization efficiency in maize-soybean relay strip intercropping system. Acta Ecologica Sinica, 2015, 35(13): 4473-4482. (in Chinese)

[28] 刘小明, 雍太文, 苏本营, 刘文钰, 周丽, 宋春, 杨峰, 王小春, 杨文钰. 减量施氮对玉米-大豆套作系统中作物产量的影响. 作物学报, 2014, 40( 9): 1629-1638.

LIU X M, YONG T W, SU B Y, LIU W Y, ZHOU L, SONG C, YANG F, WANG X C, YANG W Y. Effect of reduced N application on crop yield in maize-soybean intercropping system. Acta Agronomica Sinica, 2014, 40(9): 1629-1638. (in Chinese)

[29] 毛璐, 宋春, 徐敏, 雍太文, 王小春, 杨文钰. 栽培模式及施肥对玉米和大豆根际土壤磷素有效性的影响. 中国生态农业学报, 2015, 23(12): 1502-1510.

MAO L, SONG C, XU M, YONG T W, WANG X C, YANG W Y. Effects of cropping patterns and fertilization on rizosphere soil phosphorus availability of maize/soybean relay intercropping system. Chinese Journal of Eco-Agriculture, 2015, 23(12): 1502-1510. (in Chinese)

[30] 宋亚娜, 林智敏, 林艳. 氮肥对稻田土壤反硝化细菌群落结构和丰度的影响. 中国生态农业学报, 2012, 20 (1): 7-12.

SONG Y N, LIN Z M, LIN Y. Response of denitrifying bacteria community structure and abundance to nitrogen in paddy fields. Chinese Journal of Eco-Agriculture, 2012, 20(1): 7-12. (in Chinese)

[31] 武传东, 闫倩, 辛亮, 王保莉, 曲东. 长期施用氮肥和磷肥对渭北旱塬土壤中氨氧化古菌多样性的影响. 农业环境科学学报, 2012, 31(4): 743-749.

WU C D, YAN Q, XIN L, WANG B L, QU D. Effects of long-term nitrogen and phosphate fertilization on diversity of ammonia- oxidizing archaea in dry highland soil of Loess Plateau, China. Journal of Agro-Environment Science, 2012, 31(4): 743-749. (in Chinese)

[32] 刘骁蒨, 涂仕华, 孙锡发, 辜运富, 张先琴, 张小平. 秸秆还田与施肥对稻田土壤微生物生物量及固氮菌群落结构的影响, 生态学报, 2013, 33(17): 5210-5218.

LIU Y Q, TU S H, SUN X F, GU Y F, ZHANG X Q, ZHANG X P. Effect of different fertilizer combinations and straw return on microbial biomass and nitrogen-fixing bacteria community in a paddy soil. Acta Ecologica Sinica, 2013, 33(17): 5210-5218. (in Chinese)

[33] 周德庆. 微生物学实验教程. 北京: 高等教育出版社, 2013: 109-121, 272- 375.

ZHOU D Q. Microbiological Experiment Teaching. Beijing: Higher Education Press, 2013: 109-121, 272-375. (in Chinese)

[34] 程占冰, 杨江科, 李鹤, 朱兵, 陈相军, 闫云军. 淡水富营养型湖泊沉积物亚硝酸还原酶(nirS)的多样性和系统发育. 微生物学报, 2011, 51(5): 667-675.

CHENG Z B, YANG J K, LI H, ZHU B, CHEN X J, YAN Y J. Phylogeny diversity of the nitrite reductase gene (nirS) in the sediments of the eutrophic East Lake, Wu han. Acta Microbiologica Sinica, 2011, 51(5): 667-675. (in Chinese)

[35] 孟庆英, 张春峰, 于忠和, 贾会彬, 朱宝国, 王囡囡. 根瘤茵对大豆根际土壤微生物及大豆农艺性状的影响. 大豆科学, 2012, 31(3): 498-503.

MENG Q Y, ZHENG C F, YU Z H, JIA H B, ZHU B G, WANG N N. Effects of rhizobia on rhizosphere soil microoganisms and agronomic characters of soybean. Soybean Science, 2012, 31(3): 498-503. (in Chinese)

[36] HINSINGER P, BETENCOURT E, BERNARD L. P for two, sharing a scarce resource: Soil phosphorus acquisition in the rhizosphere of intercropped species. Plant Physiology, 2011, 156(3): 1078-1086.

[37] 雍太文, 杨文钰, 向达兵, 陈小容, 万燕. 小麦/玉米/大豆套作的产量、氮营养表现及其种间竞争力的评定. 草业学报, 2012, 21(1): 50-58.

YONG T W, YANG W Y, XIANG D B, CHEN X R, WAN Y. Production and N nutrient performance of wheat-maize-soybean relay strip intercropping system and evaluation of interspecies competition. Acta Prataculturae Sinica, 2012, 21(1): 50-58.(in Chinese)

[38] 周海霞, 刘其友, 郑经堂. 海洋微生物固氮作用研究进展. 化学与生物工程, 2013, 30(11): 5-9.

ZHOU H X, LIU Q Y, ZHENG J T. Research progress on marine microbial nitrogen fixation. Chemistry & Bioengineering, 2013, 30(11): 5-9. (in Chinese)

[39] 于亚军, 朱波, 王小国, 项虹艳, 郑循华. 成都平原水稻一油菜轮作系统氧化亚氮排放. 应用生态学报, 2008, 19(6): 1277-1282.

YU Y J, ZHU B, WANG X G, XIANG H Y, ZHENG X H. N₂O emission from rice-rapeseed rotation system in Chengdu Plain of Sichun Basin. Chinese Journal of Applied Ecology, 2008, 19(6): 1277-1282. (in Chinese)