Turn off MathJax
Article Contents
Article Navigation >  Journal of Zhejiang A&F University  > 2026  > Accepted Manuscript

BAO Daiqin, LIU Guoqun, WEN Jun, et al. Effects of combined application of green manure and bio-organic fertilizer on soil arbuscular mycorrhizal fungal community structure and Citrus reticulata fruit quality[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−13 doi:  10.11833/j.issn.2095-0756.20260182
Citation: BAO Daiqin, LIU Guoqun, WEN Jun, et al. Effects of combined application of green manure and bio-organic fertilizer on soil arbuscular mycorrhizal fungal community structure and Citrus reticulata fruit quality[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−13 doi:  10.11833/j.issn.2095-0756.20260182
shu

OnlineFirst articles are published online before they appear in a regular issue of the journal. Please find and download the full texts via CNKI.

Effects of combined application of green manure and bio-organic fertilizer on soil arbuscular mycorrhizal fungal community structure and Citrus reticulata fruit quality

DOI: 10.11833/j.issn.2095-0756.20260182
  • 1.

    College of Environment and Resources/College of Carbon Neutrality, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China

  • 2.

    Centre of Beautiful Countryside Construction, Kecheng District, Quzhou 324000, Zhejiang, China

  • Received Date: 2026-03-05
  • Accepted Date: 2026-06-12
  • Rev Recd Date: 2026-06-09
  •   Objective  The aim of this study is to explore the effects of different green manure and organic fertilizer combination treatments on the structure of soil arbuscular mycorrhizal (AM) fungal communities and Citrus reticulata fruit quality through field experiments, so as to provide a scientific basis for sustainable green C. reticulata production.   Method  In a field experiment in the C. reticulata orchard of Quzhou, Zhejiang Province, four green manure treatments were set up: clean tillage (ck), Vulpia myuros, Trifolium repens, and Astragalus sinicus, each paired with compound fertilizer (CF), commercial organic fertilizer (COF), and bio-organic fertilizer (BOF). After two years, fruit quality, soil chemical properties, enzyme activities, and AM fungal spore density were measured. AM fungal community structure was analyzed via Illumina MiSeq sequencing, and the community stability was assessed by co-occurrence network analysis.   Result  Compared with the application of ck combined with CF, the treatment of V. myuros combined with BOF significantly increased fruit soluble solids-to-titratable acidity ratio and vitamin C content by 73.6% and 16.6% (P＜0.05), respectively. At the same time, soil total nitrogen, easily extractable glomalin and AM fungal spore density significantly increased (P＜0.05). After the combined application of green manure and organic fertilizer, the dominant genera shifted to Glomus, Claroideoglomus and Paraglomus. The combination of green manure and organic fertilizer enhanced AM fungal network modularity, with a modularity index of 0.844 for the treatment of V. myuros with BOF.   Conclusion  The combined application of green manure (especially V. myuros) and bio-organic fertilizer can improve soil nutrient and glomalin contents, optimize AM fungal community structure and increase network modularity, and promote C. reticulata fruit quality. [Ch, 7 fig. 4 tab. 58 ref.]
  • [1] SHI Luyi, XU Rui, WU Qifeng, TONG Xiaodong, ZHANG Linling, MA Xingcong, QIN Hua.  Short-term response characteristics of soil bacterial community in open-field vegetable soils to combined application of organic fertilizer and soil conditioner . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20250403
    [2] LI Jiaxuan, CHEN Yuewen, ZHOU Tie, SHI Jun, YANG Junfeng, LU Xiaopeng, LI Feifei.  Genome-wide identification and drought stress response analysis of the OSCA gene family in Poncirus trifoliata . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20250103
    [3] YE Xiaoling, ZHAO Yuhong, JIANG Nan, ZHANG Min, ZHANG Chi.  Fertility effect of E3 ubiquitin ligase gene CsRNF217 in transgenic tobacco plants . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20220710
    [4] CHEN Jiayin, HUANG Chengpeng, ZHENG Mengqi, LI Zihong, WANG Jiayu, JIANG Peikun.  Effects of substituting organic fertilizer and biochar-based fertilizer instead of chemical fertilizer on nitrogen and phosphorus runoff loss in sweet potato sloping farmland . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20220360
    [5] PENG Sili, ZHANG Xin, WU Renjie, CAI Yanjiang, XING Wei, GE Zhiwei, MAO Lingfeng.  Seasonal dynamic responses of soil arbuscular mycorrhizal fungal community to nitrogen additions in a poplar plantation . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20220640
    [6] GAO Ning, XING Yijing, XIONG Rui, SHI Wenhui.  Mechanisms of plant P acquisition coordinated by arbuscular mycorrhizal fungi and phosphate-solubilizing bacteria . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20220765
    [7] REN Yi, JIANG Peikun, LU Changgen, SHAO Jianjun, ZHOU Xue’e, CHEN Junhui.  Effects of biochar-based fertilizer and organic fertilizer substituting chemical fertilizer partially on soil microbial abundances and enzyme activities . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20210619
    [8] WU Jindong, HE Yong, ZHU Zhujun.  Effects of partial substitution of chemical fertilizer with organic fertilizer on growth and quality of Solanum melongena in open field . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20200767
    [9] FANG Chenlu, JIAN Yongqi, WU Jiasen, ZHANG Yan, LU Changgen, SHAO Jianjun, GUO Feifei, JIANG Peikun.  Response of nitrogen and phosphorus uptake and runoff loss in single cropping rice to different fertilization treatments . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20200724
    [10] CHEN Limei, LI Xiaoying, LI Junlong, LIANG Zhi, SHI Liangtao.  Effects of combined application of bamboo charcoal and organic fertilizer on soil fertility and growth of Brassica oleracea var. capitata f. rubra . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.20200723
    [11] WANG Yingying, ZHAO Bing, LI Ying.  Heat resistance of Rhododendron with arbuscular mycorrhizal fungi . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.2019.04.013
    [12] QU Minghua, YU Yuanchun, LI Sheng, ZHANG Jinchi.  Advances in research on activation of mineral nutrients by arbuscular mycorrhizal fungi . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.2019.02.022
    [13] CHEN Lulu, LEI Niya.  Plant-soil feedback with arbuscular mycorrhizal fungi colonization and biomass of two introduced tree species . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.2018.03.005
    [14] ZHOU Ziqiu, LU Yuanyuan, FAN Weiqing, YE Huiqun, WU Lidong.  Effects of fertilizer on physical and mechanical properties of 5 years old Phyllostachys edulis . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.2013.05.015
    [15] WANG Guoli, AN Huaming, QIN Qiaoping, LI Mengjiao, LIU Zhenzhen, CHEN Jiaying, ZHOU Qian, ZHANG Lanlan.  Cloning and sequence analysis of fruit ripening-specific gene CsPMEI/InvI from citrus . Journal of Zhejiang A&F University, doi: 10.11833/j.issn.2095-0756.2013.03.005
    [16] WANG Min-hua, ZHOU Jing, CUI Jian.  Effects of red soil moisture on leaves growth and nutrient accumulation of Citrus unshiu . Journal of Zhejiang A&F University,
    [17] XU Qiu-fang, JIANG Pei-kun, LU Yi-tong.  Soil microbial diversity with different fertilizer types and rates in a Phyllostachys praecox stand . Journal of Zhejiang A&F University,
    [18] JIANG Pei-kun, XUQiu-fang, WUQi-feng, WUJia-sen.  Effects of fertilization on soil properties under Castanea mollissima plantation . Journal of Zhejiang A&F University,
    [19] YANG Fang, WU Jia-sen, QIAN Xin-biao, WU Li-jun.  Dynamic changes of soil microbial biomass carbon in Phyllostachys praecox stand with different fertilizers . Journal of Zhejiang A&F University,
    [20] LIU Ai-qin, MA Xiang-qing, HE Zhi-ying.  Effect of intercorpping green manure crops on degraded soil improvement of Chinese fir plantation after control burning . Journal of Zhejiang A&F University,
  • [1]
    ZHANG Xian, HU Xizhou, PENG Xitian, et al. Determination of 19 organophosphorus pesticide residues in Citrus reticulata peel based on QuEChERS-GC method[J]. Hubei Agricultural Sciences, 2020, 59(21): 147−150. DOI: 10.14088/j.cnki.issn0439-8114.2020.21.031.
    [2]
    CAI Jiaojiao, XIN Qing, LIU Wenmei. Analysis of Chinese Citrus export competitiveness under the background of “double cycle”[J]. Science and Technology of Tianjin Agriculture and Forestry, 2025(2): 33−37, 46. DOI: 10.3969/j.issn.1002-0659.2025.02.009.
    [3]
    LÜ Fenglian, SONG Jiashan, GILTRAP D, et al. Crop yield and N2O emission affected by long-term organic manure substitution fertilizer under winter wheat-summer maize cropping system[J]. Science of the Total Environment, 2020, 732: 139321. DOI: 10.1016/j.scitotenv.2020.139321.
    [4]
    WU Qiangsheng, ZOU Yingning. New advances in the research of arbuscular mycorrhizas in Citrus[J]. Acta Agriculturae Universitatis Jiangxiensis, 2014, 36(2): 279−284. DOI: 10.13836/j.jjau.2014044.
    [5]
    CHEN Rui, CUI Zeyuan, LI Zhi, et al. The effect of AM fungi on crop growth in continuous cropping soils: a review[J]. Chinese Agricultural Science Bulletin, 2024, 40(6): 1−8.
    [6]
    GAO Ning, XING Yijing, XIONG Rui, et al. Mechanisms of plant P acquisition coordinated by arbuscular mycorrhizal fungi and phosphate-solubilizing bacteria[J]. Journal of Zhejiang A&F University, 2023, 40(6): 1167−1180. DOI: 10.11833/j.issn.2095-0756.20220765.
    [7]
    OU Jing, LIU Renyang, ZHANG Ren’ai, et al. Nitrate reductase activity and N absorption of Rhododendron annae seedlings with ericoid mycorrhiza inoculation[J]. Journal of Zhejiang A&F University, 2014, 31(6): 926−931. DOI: 10.11833/j.issn.2095-0756.2014.06.015.
    [8]
    WANG Wanxiao, SHI Jincai, XIE Qiujin, et al. Nutrient exchange and regulation in arbuscular mycorrhizal symbiosis[J]. Molecular Plant, 2017, 10(9): 1147−1158. DOI: 10.1016/j.molp.2017.07.012.
    [9]
    CHEN Baodong, YU Meng, HAO Zhipeng, et al. Research progress in arbuscular mycorrhizal technology[J]. Chinese Journal of Applied Ecology, 2019, 30(3): 1035−1046. DOI: 10.13287/j.1001-9332.201903.037.
    [10]
    OMIROU M, IOANNIDES I M, EHALIOTIS C. Mycorrhizal inoculation affects arbuscular mycorrhizal diversity in watermelon roots, but leads to improved colonization and plant response under water stress only[J]. Applied Soil Ecology, 2013, 63: 112−119. DOI: 10.1016/j.apsoil.2012.09.013.
    [11]
    LÓPEZ-RÁEZ J A, CHARNIKHOVA T, GÓMEZ-ROLDÁN V, et al. Tomato strigolactones are derived from carotenoids and their biosynthesis is promoted by phosphate starvation[J]. New Phytologist, 2008, 178(4): 863−874. DOI: 10.1111/j.1469-8137.2008.02406.x.
    [12]
    YE Rongsheng. Effect of Organic Fertilier on Nutrition and Growth of Citrus[D]. Chongqing: Southwest University, 2013.
    [13]
    YE J, WANG Y, WANG Y, et al. Improvement of soil acidification and ammonium nitrogen content in tea plantations by long-term use of organic fertilizer[J]. Plant Biology, 2023, 25(6): 994−1008. DOI: 10.1111/plb.13554.
    [14]
    WANG Guocui, LI Hanting, HU Falong, et al. Application of green manure combined with synthetic nitrogen fertilizer enhances soil aggregate stability in an arid wheat cropping system[J]. Applied Soil Ecology, 2025, 206: 105849. DOI: 10.1016/j.apsoil.2024.105849.
    [15]
    WEN Jun, LIU Yidan, MA Xingcong, et al. Response of arbuscular mycorrhizal fungal communities in Citrus orchard soils to fertilization practices and their influencing factors at a regional scale[J]. Journal of Plant Nutrition and Fertilizers, 2025, 31(6): 1163−1174. DOI: 10.11674/zwyf.2024500.
    [16]
    KUZYAKOV Y. Priming effects: interactions between living and dead organic matter[J]. Soil Biology and Biochemistry, 2010, 42(9): 1363−1371. DOI: 10.1016/j.soilbio.2010.04.003.
    [17]
    LIU Yidan, WEN Jun, MA Xingcong, et al. Effects of bio-organic fertilizer on soil arbuscular mycorrhizal fungi community under different crop rotation[J]. Chinese Journal of Applied and Environmental Biology, 2025, 31(3): 486−496. DOI: 10.19675/j.cnki.1006-687x.2024.11038.
    [18]
    DENG Sufeng, YANG Shuizhi, LONG Xianyao, et al. Effect of multi-functional bio-organic fertilizer on improving alkaline soil of Citrus orchards[J]. South China Fruits, 2025, 54(3): 7−14. DOI: 10.13938/j.issn.1007-1431.20240174.
    [19]
    BAO Shidan. Soil Agrochemical Analysis[M]. 3rd ed. Beijing: China Agriculture Press, 2000.
    [20]
    SAIYA-CORK K R, SINSABAUGH R L, ZAK D R. The effects of long term nitrogen deposition on extracellular enzyme activity in an Acer saccharum forest soil[J]. Soil Biology and Biochemistry, 2002, 34(9): 1309−1315. DOI: 10.1016/S0038-0717(02)00074-3.
    [21]
    GERDEMANN J W, NICOLSON T H. Spores of mycorrhizal Endogone species extracted from soil by wet sieving and decanting[J]. Transactions of the British Mycological Society, 1963, 46(2): 235−244. DOI: 10.1016/S0007-1536(63)80079-0.
    [22]
    LEE J, LEE S, YOUNG J P W. Improved PCR primers for the detection and identification of arbuscular mycorrhizal fungi: PCR primers for arbuscular mycorrhizal fungi[J]. FEMS Microbiology Ecology, 2008, 65(2): 339−349. DOI: 10.1111/j.1574-6941.2008.00531.x.
    [23]
    HIIESALU I, PÄRTEL M, DAVISON J, et al. Species richness of arbuscular mycorrhizal fungi: associations with grassland plant richness and biomass[J]. New Phytologist, 2014, 203(1): 233−244. DOI: 10.1111/nph.12765.
    [24]
    CHEN Keyu, ZHANG Jing, MUNEER M A, et al. Plant community and soil available nutrients drive arbuscular mycorrhizal fungal community shifts during alpine meadow degradation[J]. Fungal Ecology, 2023, 62: 101211. DOI: 10.1016/j.funeco.2022.101211.
    [25]
    LIU Zherong, YANG Yi, BAI Wenke, et al. Responses of arbuscular mycorrhizal fungal communities to various artificial vegetation restoration practices and oat intercropping[J]. Southwest China Journal of Agricultural Sciences, 2025, 38(1): 171−181. DOI: 10.16213/j.cnki.scjas.2025.1.019.
    [26]
    CAO Mengyan, YE Shaoping, JIN Cheng, et al. The communities of arbuscular mycorrhizal fungi established by different winter green manures in paddy fields promote post-cropping rice production[J]. Journal of Integrative Agriculture, 2025, 24(4): 1588−1605. DOI: 10.1016/j.jia.2024.07.035.
    [27]
    DONG Ningguang, HU Guanglong, ZHANG Yunqi, et al. Effects of green-manure and tillage management on soil microbial community composition, nutrients and tree growth in a walnut orchard[J]. Scientific Reports, 2021, 11: 16882. DOI: 10.1038/s41598-021-96472-8.
    [28]
    MBUTHIA L W, ACOSTA-MARTÍNEZ V, DEBRUYN J, et al. Long term tillage, cover crop, and fertilization effects on microbial community structure, activity: implications for soil quality[J]. Soil Biology and Biochemistry, 2015, 89: 24−34. DOI: 10.1016/j.soilbio.2015.06.016.
    [29]
    ZHAO Wenjun, FENG Yu, CHEN Hua, et al. Assessment of green manure resources suitability for tobacco-growing regions in Yuxi, Yunnan Province[J]. Chinese Journal of Soil Science, 2025, 56(6): 1726−1733. DOI: 10.19336/j.cnki.trtb.2025041602.
    [30]
    LI Qingmei, WANG Hualing, ZHANG Lingling, et al. Cover cropping with white clover and ratten grass impacts soil microbial and nematode communities[J]. Plant Nutrition and Fertilizer Science, 2021, 27(6): 1055−1067. DOI: 10.11674/zwyf.20622.
    [31]
    DUAN Shilong, YAN Wenhui, FENG Gu, et al. Carbon-phosphorus reciprocal mechanism for plants to acquire nutrients through the root/mycorrhizal pathway[J]. Plant Nutrition and Fertilizer Science, 2023, 29(6): 1160−1167. DOI: 10.11674/zwyf.2022627.
    [32]
    CHEN Yiyong, LI Jianlong, ZHOU Bo, et al. Effects of intercropping with Vulpia myuros in tea plantation on soil and tea quality components[J]. Scientia Agricultura Sinica, 2023, 56(24): 4916−4929. DOI: 10.3864/j.issn.0578-1752.2023.24.010.
    [33]
    MARTIN G M, RIVERA R. Influence of mycorrhizal inoculation on green manures. Effect on the main crop. A case study: corn[J]. Cultivos Tropicales, 2015, 36: 34−50.
    [34]
    WARNER A, MOSSE B. Factors affecting the spread of vesicular mycorrhizal fungi in soil[J]. New Phytologist, 1982, 90(3): 529−536. DOI: 10.1111/j.1469-8137.1982.tb04485.x.
    [35]
    QIAN Siru, XU Ying, ZHANG Yifei, et al. Effect of AMF inoculation on reducing excessive fertilizer use[J]. Microorganisms, 2024, 12(8): 1550. DOI: 10.3390/microorganisms12081550.
    [36]
    de MIRANDA J C C, HARRIS P J. Effects of soil phosphorus on spore germination and hyphal growth of arbuscular mycorrhizal fungi[J]. New Phytologist, 1994, 128(1): 103−108. DOI: 10.1111/j.1469-8137.1994.tb03992.x.
    [37]
    LIU Jiai, SHU Aiping, SONG Weifeng, et al. Long-term organic fertilizer substitution increases rice yield by improving soil properties and regulating soil bacteria[J]. Geoderma, 2021, 404: 115287. DOI: 10.1016/j.geoderma.2021.115287.
    [38]
    PONCE M A, SCERVINO J M, ERRA-BALSELLS R, et al. Flavonoids from shoots and roots of Trifolium repens (white clover) grown in presence or absence of the arbuscular mycorrhizal fungus Glomus intraradices[J]. Phytochemistry, 2004, 65(13): 1925−1930. DOI: 10.1016/j.phytochem.2004.06.005.
    [39]
    HOU Jiaojiao, SUN Tao, YU Zhongdong, et al. Effect of endophytic fungi on growth and physiology of saline stressed Sophora japonica seedlings[J]. Journal of Zhejiang A&F University, 2017, 34(2): 294−300. DOI: 10.11833/j.issn.2095-0756.2017.02.013.
    [40]
    ZHANG Meiqing, WANG Youshan, XING Lijun. The relationship between the distribution of AM fungi and environmental factors[J]. Mycosystema, 1999, 18(1): 25−29. DOI: 10.13346/j.mycosystema.1999.01.005.
    [41]
    MA Li, CHEN Jianxin, KANG Dingxu, et al. Identification and diversity of arbuscular mycorrhizal fungi (AMF) and microorganisms of Bombax ceiba in dry-hot valley[J]. Journal of Northwest Forestry University, 2023, 38(1): 131−138. DOI: 10.3969/j. issn. 1001-7461.2023. 01.18.
    [42]
    ZHANG Cunzhi, XIANG Xingjia, YANG Teng, et al. Nitrogen fertilization reduces plant diversity by changing the diversity and stability of arbuscular mycorrhizal fungal community in a temperate steppe[J]. Science of the Total Environment, 2024, 918: 170775. DOI: 10.1016/j.scitotenv.2024.170775.
    [43]
    ZHANG Tantan, YU Lingling, SHAO Yuting, et al. Root and hyphal interactions influence N transfer by arbuscular mycorrhizal fungi in soybean/maize intercropping systems[J]. Fungal Ecology, 2023, 64: 101240. DOI: 10.1016/j.funeco.2023.101240.
    [44]
    ACIEGO PIETRI J C, BROOKES P C. Substrate inputs and pH as factors controlling microbial biomass, activity and community structure in an arable soil[J]. Soil Biology and Biochemistry, 2009, 41(7): 1396−1405. DOI: 10.1016/j.soilbio.2009.03.017.
    [45]
    MA Xiaocui, XU Xia, GENG Qinghong, et al. Global arbuscular mycorrhizal fungal diversity and abundance decreases with soil available phosphorus[J]. Global Ecology and Biogeography, 2023, 32(8): 1423−1434. DOI: 10.1111/geb.13704.
    [46]
    TAWARAYA K, SAITO M, MORIOKA M, et al. Effect of phosphate application to arbuscular mycorrhizal onion on the development and succinate dehydrogenase activity of internal hyphae[J]. Soil Science and Plant Nutrition, 1994, 40(4): 667−673. DOI: 10.1080/00380768.1994.10414306.
    [47]
    ZHANG Xiangyu, CHEN Xiaofang, LIU Min, et al. Correlation between arbuscular mycorrhiza fungi diversity and environmental factors in rotation soil of Pinellia ternate[J]. Jiangsu Agricultural Sciences, 2024, 52(2): 213−220. DOI: 10.15889/j.issn.1002-1302.2024.02.030.
    [48]
    JIN Wenhao, SHAO Shuai, CHEN Junhui, et al. Research progress in the impact of different mycorrhizal types on soil carbon cycling[J]. Journal of Zhejiang A&F University, 2021, 38(5): 953−962. DOI: 10.11833/j.issn.2095-0756.20210531.
    [49]
    GRILLI J, ROGERS T, ALLESINA S. Modularity and stability in ecological communities[J]. Nature Communications, 2016, 7: 12031. DOI: 10.1038/ncomms12031.
    [50]
    CHEN Xinyu, HU Yongqing, LÜ Junting, et al. Effects of biochar application on bacterial community structure and symbiotic network analysis in acidic purple soil[J]. Environmental Science, 2025, 46(12): 8071−8081. DOI: 10.13227/j.hjkx.202412036.
    [51]
    YE Zhencheng, LI Jing, WANG Jie, et al. Diversity and co-occurrence network modularization of bacterial communities determine soil fertility and crop yields in arid fertigation agroecosystems[J]. Biology and Fertility of Soils, 2021, 57(6): 809−824. DOI: 10.1007/s00374-021-01571-3.
    [52]
    GAO Cheng, XU Ling, MONTOYA L, et al. Co-occurrence networks reveal more complexity than community composition in resistance and resilience of microbial communities[J]. Nature Communications, 2022, 13: 3867. DOI: 10.1038/s41467-022-31343-y.
    [53]
    FENG Yu, LIANG Hai, NIE Jun, et al. Roles of microbial community and keystone taxa in rice productivity under green manuring in South China[J]. Sustainability, 2024, 16(9): 3565. DOI: 10.3390/su16093565.
    [54]
    BASTIDA F, ELDRIDGE D J, GARCÍA C, et al. Soil microbial diversity-biomass relationships are driven by soil carbon content across global biomes[J]. The ISME Journal, 2021, 15(7): 2081−2091. DOI: 10.1038/s41396-021-00906-0.
    [55]
    LIN Guigang, MCCORMACK M L, GUO Dali. Arbuscular mycorrhizal fungal effects on plant competition and community structure[J]. Journal of Ecology, 2015, 103(5): 1224−1232. DOI: 10.1111/1365-2745.12429.
    [56]
    ZHANG Yu, DONG Lingbo, SHANGGUAN Zhouping. Appropriate N addition improves soil aggregate stability through AMF and glomalin-related soil proteins in a semiarid agroecosystem[J]. Land Degradation & Development, 2023, 34(3): 710−722. DOI: 10.1002/ldr.4488.
    [57]
    LIU Yiwen, GUAN Dongxing, QIU Lixue, et al. Spatial dynamics of phosphorus mobilization by mycorrhiza[J]. Soil Biology and Biochemistry, 2025, 206: 109797. DOI: 10.1016/j.soilbio.2025.109797.
    [58]
    YANG Bo, WANG Shaojun, ZHAO Shuang, et al. Effects of arbuscular mycorrhizal fungi symbiosis on growth and photosynthetic characteristics of Fraxinus malacophylla in rocky desertification habitats[J]. Journal of Zhejiang A&F University, 2022, 39(5): 1028−1036. DOI: 10.11833/j.issn.2095-0756.20210740.
  • 加载中
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Figures(7)  / Tables(4)

Get Citation
PDF
XML

Article views(88) PDF downloads(16) Cited by()

Related
Proportional views

Effects of combined application of green manure and bio-organic fertilizer on soil arbuscular mycorrhizal fungal community structure and Citrus reticulata fruit quality

doi: 10.11833/j.issn.2095-0756.20260182
  • 1. College of Environment and Resources/College of Carbon Neutrality, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China
  • 2. Centre of Beautiful Countryside Construction, Kecheng District, Quzhou 324000, Zhejiang, China
  • Received Date: 2026-03-05
  • Accepted Date: 2026-06-12
  • Rev Recd Date: 2026-06-09

Abstract:   Objective  The aim of this study is to explore the effects of different green manure and organic fertilizer combination treatments on the structure of soil arbuscular mycorrhizal (AM) fungal communities and Citrus reticulata fruit quality through field experiments, so as to provide a scientific basis for sustainable green C. reticulata production.   Method  In a field experiment in the C. reticulata orchard of Quzhou, Zhejiang Province, four green manure treatments were set up: clean tillage (ck), Vulpia myuros, Trifolium repens, and Astragalus sinicus, each paired with compound fertilizer (CF), commercial organic fertilizer (COF), and bio-organic fertilizer (BOF). After two years, fruit quality, soil chemical properties, enzyme activities, and AM fungal spore density were measured. AM fungal community structure was analyzed via Illumina MiSeq sequencing, and the community stability was assessed by co-occurrence network analysis.   Result  Compared with the application of ck combined with CF, the treatment of V. myuros combined with BOF significantly increased fruit soluble solids-to-titratable acidity ratio and vitamin C content by 73.6% and 16.6% (P＜0.05), respectively. At the same time, soil total nitrogen, easily extractable glomalin and AM fungal spore density significantly increased (P＜0.05). After the combined application of green manure and organic fertilizer, the dominant genera shifted to Glomus, Claroideoglomus and Paraglomus. The combination of green manure and organic fertilizer enhanced AM fungal network modularity, with a modularity index of 0.844 for the treatment of V. myuros with BOF.   Conclusion  The combined application of green manure (especially V. myuros) and bio-organic fertilizer can improve soil nutrient and glomalin contents, optimize AM fungal community structure and increase network modularity, and promote C. reticulata fruit quality. [Ch, 7 fig. 4 tab. 58 ref.]

BAO Daiqin, LIU Guoqun, WEN Jun, et al. Effects of combined application of green manure and bio-organic fertilizer on soil arbuscular mycorrhizal fungal community structure and Citrus reticulata fruit quality[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−13 doi:  10.11833/j.issn.2095-0756.20260182
Citation: BAO Daiqin, LIU Guoqun, WEN Jun, et al. Effects of combined application of green manure and bio-organic fertilizer on soil arbuscular mycorrhizal fungal community structure and Citrus reticulata fruit quality[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−13 doi:  10.11833/j.issn.2095-0756.20260182
shu

    HTML

  • 柑橘Citrus reticulata是世界第一大类水果，种植面积和产量长期位居全球首位，其产业健康发展对保障优质果品供给至关重要[1−2]。由于长期集约化栽培，橘园中过量投入化肥和农药等，引发土壤酸化板结、养分失衡，土壤微生物活性减弱、生物多样性下降以及生态系统功能衰退，已严重制约柑橘产业可持续发展[3]。通过定向调控土壤有益微生物群落结构、优化提升土壤生态环境质量，可推动橘园栽培管理向绿色生态转型，进而助力柑橘产业实现提质增效与稳产优产的目标。研究发现：柑橘属于菌根依赖型植物，根毛稀少或无根毛，其养分吸收高度依赖丛枝菌根(arbuscular mycorrhizal，AM)真菌的协助[4]。丛枝菌根真菌是广泛分布于陆地生态系统中的土壤微生物，可与约80%陆生植物根系形成互惠共生关系[5]。丛枝菌根真菌菌丝网络能扩大根系的吸收范围，协助宿主获取土壤中移动性较差的磷、锌、铜等矿质元素。研究表明：接种丛枝菌根真菌可提高柑橘幼苗的磷吸收效率，促进根系发育，增强叶片光合能力，进而改善植株生物量积累与早期建植[6−7]。此外，丛枝菌根真菌可通过诱导系统抗性，竞争根际生态位等方式，增强柑橘对根腐病等土传病害及干旱、盐胁迫等非生物胁迫的抗性[8−9]。丛枝菌根真菌菌丝分泌的球囊霉素相关蛋白(glomalin-related soil protein，GRSP)，能够胶结土壤颗粒，促进水稳性团聚体的形成，从而改善土壤结构，增强孔隙度和保水能力；还可通过调节土壤酶活性，与根际微生物互作，影响土壤碳氮周转与有机质稳定[10−11]。因此，加强橘园丛枝菌根真菌多样性保育有助于提升土壤生物活性，增强生态系统稳定性与功能韧性，是实现柑橘园土壤健康管理和可持续发展的关键生物策略。

    柑橘园的绿色发展通常采用有机肥与化肥配施模式，从而实现化肥减量，果实品质提升及土壤健康[12]。增施有机肥、种植绿肥等措施已成为改良橘园土壤，提升土壤生物活性的重要途径。有机肥可改善土壤理化性质、缓解酸化，并为微生物提供碳源与养分，促进有益菌群增殖；绿肥则可改善土壤结构、活化养分并调控根际微生物群落[13−14]。单施化肥不利于丛枝菌根真菌与植物共生，而适量有机肥或绿肥配施能提高丛枝菌根真菌的α多样性，改变群落结构，并富集球囊霉属Glomus等有益菌群[15−17]。这些措施协同作用有助于恢复土壤微生态平衡，增强系统抗逆性与养分转化效率，从而为柑橘果实品质提升和柑橘产业绿色发展提供生态支撑。然而，在橘园配施有机肥和绿肥后，土壤丛枝菌根真菌群落的响应规律和机制，对柑橘品质有何影响，目前仍不清楚。

    本研究选择成熟柑橘园建立样地，设置清耕、鼠茅草Vulpia myuros、白三叶Trifolium repens、紫云英Astragalus sinicus配施复合肥、商品有机肥、生物有机肥等处理，探究绿肥和有机肥配施对柑橘果实品质、土壤酶活性及丛枝菌根真菌群落结构的影响，并探究丛枝菌根真菌群落与柑橘品质的关系，为橘园土壤健康管理，柑橘果实品质提升及产业绿色可持续发展提供理论依据。

    • 1.   材料与方法

      1.1.   试验设计及样品采集

    • 试验地位于浙江省衢州市柯城区园中园家庭农场。试验品种为‘椪柑’‘Ponkan’，大棚种植，7~8 年生，且树势相近。栽植密度为60 株·hm−2。常规种栽模式为果园清耕杂草，施用复合肥。以清耕作为对照(ck)，设置种植鼠茅草(VM)、白三叶草(TR)和紫云英(AS)等3个绿肥处理，每个处理再分别设置施常规复合肥(CF)、复合肥配施商品有机肥(COF)和复合肥配施生物有机肥(BOF)等3个施肥处理，共计12个处理，每个处理3个重复，共计36个小区；每个小区包括5株，小区随机区组排列。试验所用复合肥氮(N)∶五氧化二磷(P2O5)∶氧化钾(K2O)比例(质量比)为15∶15∶15，年施用量为1.5 kg·株−1，分 3~4 次撒施于根部周围地表；商品有机肥主要原料为牛粪、木屑和菇渣，总养分N+P2O5+K2O质量分数≥4.0%，有机质质量分数≥30%；生物有机肥主要原料为畜禽粪、菜籽饼、豆粕、核桃壳，总养分N+P2O5+K2O质量分数≥4.0%，有机质质量分数≥50%；解淀粉芽孢杆菌Bacillus amyloliquefaciens密度5×107 CFU·g−1。COF和BOF的年施用量均为10 kg·株−1，沿滴水线开沟施入。2022年8月起对所有小区进行清耕，9月底在树下和行间种植绿肥，播种量控制在11.25 kg·hm−2左右，次年4月初绿肥全部翻压入土。

      试验开展2 a后，于2024年9月底第3轮绿肥种植前，在每个小区用直径5 cm的不锈钢土钻采集来自5株柑橘树树冠水滴线内、深度为20 cm的土芯样品，总计30个，充分混匀，置于冰盒内带回实验室。一部分土壤样品保存于−80 ℃冰箱，用于土壤总DNA的提取和酶活性的测定；另一部分土壤样品经自然风干后，用于测定土壤化学性质及丛枝菌根真菌孢子密度。待12月果实成熟期，分别于每个小区每株柑橘树冠外围中部位置随机采集2个柑橘果实，混合后带回实验室测定品质与养分指标。柑橘的可滴定酸参照邓素枫等[18]的方法，采用浓度为0.1 mol·L−1氢氧化钠溶液滴定法测定。柑橘单果质量用百分之一精度的电子天平测定；可溶性固形物使用ATAGO PAL-3型手持折光仪测定；固酸比通过“可溶性固形物/可滴定酸”计算获得；维生素C质量分数则通过2,6 -二氯酚靛酚滴定法测定。

      • 1.2.   土壤化学性质与酶活性分析

    • 土壤pH与养分参照《土壤农业化学分析方法》测定[19]。pH按照1.0∶2.5的土水质量比充分搅拌后，采用玻璃电极法测定；有机碳通过重铬酸钾氧化-硫酸外加热法测定；全氮采用硫酸-加速剂消解，凯氏定氮法测定；碱解氮采用碱解扩散法测定；有效磷采用碳酸氢钠浸提法处理后，通过钼锑抗比色法测定；有效钾采用乙酸铵浸提法处理后，通过火焰光度计测定；易提取态球囊霉素(EE-GRSP)先以柠檬酸钠溶液浸提，再通过考马斯亮蓝法显色，最后在分光光度计上完成测定。采用SAIYA-CORK等[20]的荧光微孔板检测技术测定β-葡萄糖苷酶(BG)、N-乙酰-β-氨基葡萄糖苷酶(NAG)和酸性磷酸酶(PHOS)等3种土壤酶的活性。主要步骤如下：将2 g新鲜土壤加入醋酸钠缓冲液中，以180 r·min−1的转速震荡30 min后，利用八通道移液器分3次将土壤悬液转移至96孔板中，同时设置空白、标准对照物质和特定底物的对照。加样完成后，将96孔板在25 ℃下恒温培养3 h，然后使用Biotek®Synergy H1型酶标仪(Biotek)在365 nm激发光和450 nm发射光下读取水解酶的荧光值或吸光值。

      • 1.3.   土壤丛枝菌根真菌孢子密度测定、总DNA提取和高通量测序分析

    • 先将土样自然风干，再采用湿筛倾析法进行操作[21]。具体操作方法如下：先称取10 g土样置于烧杯中，加入少量水浸泡20~30 min至充分润湿；加入适量自来水，用玻璃棒快速搅悬土液，静置15 s待粗颗粒沉淀，将土悬液缓慢依次过60、120、400目网筛，于缓水流下冲洗至滤出液澄清，收集400目筛面上含孢子的悬液至培养皿，于显微镜下观察并计数。统计以每1 g土壤中丛枝菌根真菌孢子数量作为孢子密度。土壤总DNA的提取采用MoBio PowerSoil® DNA Isolation Kit试剂盒(MoBio Laboratories)，称取0.5 g的−80 ℃冻干土样，按说明书提取DNA。提取后的DNA片段大小经1%的琼脂糖凝胶电泳检测，并用微量分光光度计(NanoDrop ND-1000，Thermo Scientific)进行浓度测定，合格后送至上海美吉生物医药科技有限公司测定。

      采用巢式聚合酶链式反应(PCR)扩增丛枝菌根真菌，第1轮引物为AML1F(5′-ATCAACTTTCGATGGTAGAGATAGA-3′)和AML2R(5′-GAACCCAAACACTTTGGTTTCC-3′)，第2轮引物为AMV4-5NF(5′-AAGCTCGTAGTTGAATTTCG-3′)和AMDGR(5′-CCCAACTATCCCTATTAATCAT-3′)。扩增产物使用Illumina MiSeqTM平台进行测序[22]。原始fastq文件由测序数据质控软件Trimmomatic进行质量过滤，FLASH软件进行合并。使用USEARCH11的“uparse”算法将相似性序列(＞97%)聚类为操作分类单元(OTU)，每个OTU对应于1条代表序列。将过滤所得代表序列在MaarjAM数据库进行比对，选择标准：①一致性≥97%；②覆盖度≥97%；③期望值＜1e−50。将不符合的序列再与美国生物技术信息中心(NCBI)进行BLAST筛选，选择标准：①一致性≥90%；②覆盖度≥90%；③期望值＜1e−50，剔除不符合标准的OTU (非丛枝菌根真菌)。基于RDP 11.5数据库，采用RDP classifier方法对OTU进行物种注释。

      • 1.4.   数据统计分析

    • 采用Origin 2021软件进行绘图，利用SPSS 26软件进行统计分析，利用单因素方差分析(one-way ANOVA)和邓肯法(Duncan)对柑橘果实品质和土壤养分及微生物多样性进行差异显著性检验(P＜0.05)。土壤微生物群落结构利用上海美吉生物医药科技有限公司提供的i-Sanger平台(https://www.majorbio.com/)进行分析；使用R4.4.0的“linkET”包对丛枝菌根真菌群落与土壤化学性质间的相关关系进行Mantel检验，使用“randomForest”包进行随机森林模型分析；为区分并解析绿肥类型和施肥方式的独立效应，阐明不同处理下丛枝菌根真菌群落的种间互作模式、生态位分化特征及共现网络稳定性，本研究分别构建不同绿肥处理与不同施肥方式下的丛枝菌根真菌群落共现网络，并采用Gephi 0.9.2 (https://gephi.org)进行网络可视化与分析。

    2.   结果与分析

      2.1.   土壤化学性质及酶活性

    • 如表1所示：种植绿肥、施肥及其交互作用对柑橘土壤pH、有机碳、全氮、有效磷和速效钾影响显著(P＜0.05)。在清耕和鼠茅草处理中，商品有机肥施用显著提高土壤pH (P＜0.05)，缓解了土壤酸化。在绿肥处理中，生物有机肥施用反而降低了土壤有机碳质量分数，尤其在种植鼠茅草处理下，与常规施肥处理达到显著差异(P＜0.05)。除种植白三叶处理外，清耕及其他绿肥处理配施商品有机肥，可普遍提高土壤全氮和碱解氮质量分数。与清耕相比，种植鼠茅草和白三叶处理土壤有效磷和速效钾有降低趋势，但是与常规施肥相比，配施有机肥处理土壤速效磷、土壤速效钾质量分数未呈现统一的变化规律。种植绿肥和施肥均显著影响柑橘土壤易提取态球囊霉素质量分数(P＜0.05)。

      处理 pH 有机碳/(g·kg−1) 全氮/(g·kg−1) 碱解氮/(mg·kg−1) 有效磷/(mg·kg−1)
      ck CF 5.18±0.03 Aa 22.50±1.01 Bb 0.93±0.06 Aa 40.30±4.37 ABa 67.60±4.26 Bc
      COF 7.18±0.22 Bb 17.60±1.09 BCa 1.66±0.07 Ab 75.00±6.25 Bb 54.00±2.26 Ab
      BOF 5.66±0.08 Ba 20.90±0.55 Cb 1.49±0.13 Bb 46.70±6.17 Ba 36.80±1.59 Ba
      VM CF 6.81±0.23 Bb 28.80±2.83 Cb 1.08±0.11 Aa 20.70±4.06 Aa 41.20±1.21 Ab
      COF 7.43±0.13 Bb 21.80±0.74 Cab 2.11±0.07 Bb 97.70±4.33 Cb 53.10±2.46 Ac
      BOF 5.32±0.31 ABa 15.20±2.08 ABa 0.95±0.10 Aa 25.30±6.12 Aa 18.60±2.96 Aa
      TR CF 6.47±0.24 Ba 17.90±1.88 ABa 1.77±0.13 Ba 67.70±8.41 Cb 42.60±3.19 Aa
      COF 6.12±0.07 Aa 14.40±1.96 ABa 1.40±0.10 Aa 30.30±4.67 Aa 62.50±4.26 Ab
      BOF 6.41±0.04 Ca 20.10±2.01 BCa 1.41±0.10 Ba 32.70±4.67 ABa 86.50±1.57 Dc
      AS CF 7.18±0.09 Cc 14.50±1.54 Aa 1.74±0.08 Ba 43.70±8.25 Ba 73.60±4.44 Bb
      COF 6.31±0.17 Ab 12.40±1.24 Aa 2.16±0.10 Ba 69.30±1.76 Bb 54.20±4.53 Aa
      BOF 4.92±0.25 Aa 13.90±1.48 Aa 1.97±0.15 Ca 49.30±1.45 Ba 57.80±2.98 Ca
      GM ** ** ** ** *
      F ** ** ** ns **
      GM×F ** ** ** ** **
      处理 速效钾/(mg·kg−1) 易提取态球囊霉素/
      (g·kg−1)      		 β-葡萄糖苷酶/
      (nmol·g−1·h−1)      		 N-乙酰-β-氨基葡萄糖苷酶/
      (nmol·g−1·h−1)      		 酸性磷酸酶/
      (nmol·g−1·h−1)
      ck CF 697.00±21.50 Bb 2.09±0.20 Aa 32.13±6.30 Aa 25.88±7.14 Aa 108.86±10.65 Aa
      COF 598.00±23.00 Aa 3.19±0.06 Ab 37.33±2.60 Aa 21.33±1.76 Aa 94.60±3.56 Aa
      BOF 625.00±16.60 Ba 3.48±0.08 Ab 30.67±1.45 Aa 15.33±0.88 Aa 110.00±3.61 Aa
      VM CF 729.00±29.40 Bc 3.24±0.06 Ba 34.58±7.00 Aa 24.10±4.59 Aa 159.75±53.78 Aa
      COF 538.00±66.40 Ab 3.86±0.11 BCb 41.02±4.50 Aa 17.31±2.67 ABa 101.03±12.35 Aa
      BOF 344.00±32.00 Aa 4.30±0.10 Bc 15.29±1.83 Ba 20.31±4.86 Aa 105.12±17.48 Aa
      TR CF 526.00±11.10 Aa 3.36±0.13 Ba 36.70±6.43 Aa 14.88±3.22 Aa 126.64±29.36 Aa
      COF 580.00±27.50 Aa 3.64±0.23 ABa 28.36±8.36 ABa 22.21±5.13 Aa 119.22±8.17 Aa
      BOF 805.00±90.30 Db 3.99±0.23 Ba 22.51±5.84 ABa 22.35±4.96 Aa 140.56±4.50 Aa
      AS CF 598.00±38.40 Aa 3.66±0.27 Ba 28.90±4.77 Aa 11.55±0.54 Aa 100.40±7.37 Aa
      COF 534.00±38.40 Aa 4.15±0.14 Ca 15.79±5.27 Ba 7.93±1.57 Ba 50.55±11.08 Ba
      BOF 733.00±30.50 Cb 4.17±0.10 Ba 14.75±1.74 Ba 17.56±6.33 Aa 105.84±11.30 Aa
      GM ** ** * * *
      F ** ** ** * *
      GM×F ** ns ns * *
      　　说明：GM. 种植绿肥；F. 施肥；ck. 清耕；VM. 鼠茅草；TR. 白三叶；AS. 紫云英；CF. 复合肥；COF. 商品有机肥；BOF. 生物有机肥。不同大写字母表示在同一施肥处理下不同绿肥处理间差异显著(P＜0.05)，不同小写字母表示在同种绿肥处理下不同施肥处理间差异显著(P＜0.05)。*、**分别表示在0.05和0.01水平上差异显著。ns表示差异不显著。

      Table 1.  Soil chemical properties and enzyme activities under different fertilization treatments

      与清耕配相比，种植3种绿肥均能显著提高易提取态球囊霉素质量分数(P＜0.05)；在清耕和鼠茅草处理中配施商品有机肥和生物有机肥亦均可显著提升其质量分数(P＜0.05)。种植绿肥和施肥对土壤β-葡萄糖苷酶活性具有显著影响(P＜0.05)，但两者的交互作用无显著影响。与清耕处理相比，种植紫云英处理在施用商品有机肥的情况下显著降低了土壤β-葡萄糖苷酶活性(P＜0.05)；在施用生物有机肥的情况下，种植鼠茅草和紫云英处理均显著降低了土壤β-葡萄糖苷酶活性(P＜0.05)。种植绿肥、施肥以及两者的交互作用对土壤N-乙酰-β-氨基葡萄糖苷酶和酸性磷酸酶活性均无显著影响。

      • 2.2.   土壤丛枝菌根真菌孢子密度与群落结构

    • 从图1可看出：在清耕条件下，相较于施用复合肥，施用生物有机肥可显著提高柑橘土壤孢子密度(P＜0.05)；在鼠茅草处理中，同样是施用生物有机肥较复合肥显著提升柑橘土壤孢子密度(P＜0.05)；在施用复合肥的前提下，与清耕相比，种植鼠茅草和白三叶均能显著提高柑橘土壤孢子密度(P＜0.05)，而紫云英处理对孢子密度无显著影响。不同施肥处理的柑橘园土壤中，近明球囊霉属Claroideglomus、球囊霉属Glomus和类球囊霉属Paraglomus均为主要的丛枝菌根真菌属(图2)。种植绿肥处理对近明球囊霉属和球囊霉属的相对丰度具有显著影响(P＜0.05)，而施肥处理仅对近明球囊霉属的相对丰度影响显著(P＜0.05，图2A)。此外，种植绿肥与有机肥配施仅对球囊霉属的相对丰度表现出显著的交互作用(P＜0.05，图2B)。在清耕处理中，与施用复合肥相比，施用有机肥(商品有机肥、生物有机肥)的2个处理均显著降低了球囊霉属的相对丰度(P＜0.05)。在施用复合肥的条件下，与清耕处理相比，种植绿肥处理显著降低了球囊霉属的相对丰度(P＜0.05，图2B)。在鼠茅草处理中，与施用复合肥相比，施用商品有机肥显著降低了类球囊霉属的相对丰度(P＜0.05，图2C)。

      Figure 1.  Effect of fertilization and green manure effects on soil spore density in citrus orchards

      Figure 2.  Relative abundance of AM fungi at genus level

      基于Bray-Curtis距离矩阵的非度量多维尺度分析(NMDS)结果表明：种植不同的绿肥处理土壤丛枝菌根真菌群落之间没有明显区分，但种植白三叶处理土壤丛枝菌根真菌群落与清耕对照在第1轴上具有差异(图3A)。不同施肥处理对丛枝菌根真菌群落结构没有显著影响(图3B)。利用共现网络分析探究种植绿肥处理(图4)和不同施肥方式下(图5)柑橘土壤丛枝菌根真菌群落的共现网络关系。结果表明：种植绿肥处理(鼠茅草、白三叶、紫云英)较清耕处理相比，降低了网络节点和边的数量，但网络模块化程度明显提高(表2)。在不同施肥处理中，与施用复合肥处理相比，虽然有机肥处理(商品有机肥、生物有机肥)降低了共现网络的节点和边的数量，但提高了网络模块化程度并缩短了网路直径，施用有机肥(商品有机肥、生物有机肥)的群落网络结构更为复杂(表3)。

      Figure 3.  NMDS of AM fungi community composition at genus level according to green manure treatment and fertilization treatment

      Figure 4.  Soil AM fungal co-occurrence network under different green manure treatment

      Figure 5.  Soil AM fungal co-occurrence network under different fertilization treatments

      处理 节点 边 模块化 网络直径 网络密度 平均加权度
      清耕 39 97 0.536 3 0.131 4.811
      鼠茅草 28 34 0.704 2 0.090 2.401
      白三叶 35 43 0.857 2 0.072 2.439
      紫云英 14 10 0.778 1 0.110 1.417

      Table 2.  Topological parameters of soil AM fungi co-occurrence network　　　

      处理 节点 边 模块化 网络直径 网络密度 平均加权度
      复合肥 40 70 0.525 4 0.090 3.182
      商品有机肥 20 22 0.731 1 0.116 2.195
      生物有机肥 25 21 0.844 1 0.070 1.680

      Table 3.  Topological parameters of soil AM fungi co-occurrence network　　　

      • 2.3.   柑橘果实品质及与土壤、丛枝菌根真菌的关联分析

    • 由表4可知：在各绿肥处理下，施生物有机肥可显著增加柑橘单果质量(P＜0.05)。果实可滴定酸受栽培模式与施肥措施显著影响，其中清耕配施常规复合肥处理可滴定酸质量分数高达3.13%±0.12%，而鼠茅草配施商品有机肥处理仅为1.91%±0.07%，酸度降幅显著，更贴合优质柑橘酸甜适口的感官品质标准。整体而言，施用商品有机肥和生物有机肥显著降低果实可滴定酸，且鼠茅草、紫云英处理的可滴定酸质量分数显著低于清耕与白三叶处理(P＜0.05)。绿肥和有机肥配施对果实可溶性固形物没有显著影响，但可显著提高果实固酸比(P＜0.05)，且两者具有显著的交互效应。绿肥配施有机肥可显著提升果实维生素C质量分数(P＜0.05)，其中鼠茅草、白三叶配施有机肥效果最优。

      处理 单果质量/g 可滴定酸/% 可溶性固形物/% 固酸比 维生素C/
      (mg·100 g−1)
      ck CF 151.00±6.57 Aa 3.13±0.12 Aa 12.30±0.57 Aab 3.94±0.23 Aa 15.70±0.20 Aa
      COF 182.00±9.80 Aa 2.80±0.13 Ba 11.40±0.29 Aa 4.09±0.15 Aa 16.10±0.38 Aab
      BOF 192.00±21.0 Aa 2.75±0.23 Ba 13.60±0.81 Ab 4.98±0.24 Ab 17.10±0.26 Ab
      VM CF 151.00±1.46 Aa 2.67±0.12 Ab 11.10±0.23 Aa 4.16±0.13 ABa 18.00±0.46 Ba
      COF 169.00±9.64 Aa 2.20±0.07 Aa 12.20±0.07 Bb 5.56±0.12 Bb 18.10±0.12 Ba
      BOF 209.00±15.7 Ab 1.91±0.07 Aa 13.00±0.13 Ac 6.84±0.26 Bc 18.30±0.39 ABa
      TR CF 160.00±8.39 Aa 2.78±0.17 Aa 12.70±0.60 Aa 4.58±0.09 Ba 18.10±0.80 Ba
      COF 170.00±7.75 Aa 2.94±0.09 Ba 13.10±0.21 Ca 4.46±0.11 Aa 18.10±0.20 Ba
      BOF 209.00±13.8 Ab 2.73±0.09 Ba 13.00±1.17 Aa 4.76±0.32 Aa 17.70±0.56 ABa
      AS CF 171.00±8.68 Aa 2.79±0.28 Ab 11.70±0.93 Aa 4.23±0.24 ABa 17.20±0.15 ABa
      COF 184.00±8.50 Aa 2.10±0.10 Aa 12.60±0.18 BCa 6.06±0.37 Bb 18.30±0.46 Bb
      BOF 246.00±12.2 Ab 2.39±0.08 Bab 11.80±0.18 Aa 4.94±0.20 Aa 18.60±0.15 Bb
      GM ** ** ns ** ns
      F * ** ns ** **
      GM×F ns ns ns ** ns
      　　说明：GM. 绿肥；F. 施肥；ck. 清耕；VM. 鼠茅草；TR. 白三叶；AS. 紫云英；CF. 复合肥；COF. 商品有机肥；BOF. 生物有机肥。不同大写字母表示在同一施肥处理下不同绿肥处理间差异显著(P＜0.05)，不同小写字母表示在同种绿肥处理下不同施肥处理间差异显著(P＜0.05)。*、**分别表示在0.05和0.01水平上差异显著。ns表示差异不显著。

      Table 4.  Quality of citrus under different fertilization treatments

      从图6可以看出：果实固酸比、果实维生素C质量分数以及单果质量均与土壤有机碳质量分数呈显著负相关(P＜0.05)，而与易提取态球囊霉素呈显著(P＜0.05)或极显著正相关(P＜0.01)。此外，果实固酸比与土壤速效钾、有效磷呈显著负相关(P＜0.05)；单果质量则与近明球囊霉属Claroideoglomus的相对丰度呈极显著正相关，与类球囊霉属的相对丰度呈显著负相关(P＜0.05)。由图7可以看出，易提取态球囊霉素与近明球囊霉属的相对丰度呈显著正相关(P＜0.05)，而与类球囊霉属的相对丰度呈极显著负相关(P＜0.01)，土壤有机碳则呈相反的相关趋势。同时，土壤全氮与球囊霉属和类球囊霉属的相对丰度呈极显著负相关(P＜0.01)。土壤碱解氮仅与球囊霉属的相对丰度呈显著负相关(P＜0.05)，土壤pH与近明球囊霉属的相对丰度呈显著负相关(P＜0.05)。

      Figure 6.  Correlation analysis of citrus fruit weight and quality with soil chemical properties, AM fungal spore density and relative abundance at genus level

      Figure 7.  correlation analysis between soil chemical properties and AM fungi

    3.   讨论

      3.1.   绿肥种类与施肥方式是塑造丛枝菌根真菌群落结构的关键因素

    • 植物群落多样性和生物量决定了丛枝菌根真菌的多样性，驱动丛枝菌根真菌群落组成，且植物种类对丛枝菌根真菌群落具有显著影响[23−25]。丛枝菌根真菌在绿肥根中定殖后，可通过菌丝产生孢子，进而促进土壤中丛枝菌根真菌孢子密度的积累[26]。本研究结果也指出：鼠茅草和白三叶配施常规复合肥的丛枝菌根真菌孢子密度显著高于清耕组，而紫云英组无显著差异。这可能与不同种绿肥向土壤中释放的化合物种类不一致，导致相应处理的土壤微生物群落结构变化差异显著有关[27]。相关研究指出：氮有效性的提高对丛枝菌根真菌的相对丰度呈显著负相关。本研究中由于紫云英的强固氮能力而增加了土壤的氮输入，土壤丛枝菌根真菌孢子密度低于清耕对照组[28]。本研究表明：配施生物有机肥时，白三叶处理土壤有效磷较鼠茅草处理高出近4倍，这可能与不同种绿肥的特性相关[29]。鼠茅草须根密集、磷活化能力强，且与丛枝菌根真菌共生效率高，可加速土壤有效磷吸收消耗；白三叶以固氮为主，磷活化及丛枝菌根真菌共生能力较弱，土壤有效磷易累积，进而造成两者差异显著[30−31]。本研究结果显示：不同种绿肥对柑橘土壤中近明球囊霉属和球囊霉属相对丰度影响显著，其中鼠茅草配施商品有机肥处理显著降低了近明球囊霉属相对丰度，这可能与种植鼠茅草降低了土壤球囊菌门Glomeromycota相对丰度相关[32]。球囊霉属具有广泛的功能分布，主要分布在高、中肥力生态系统中，具有极强的效率和竞争力，且更易于定殖于白三叶根系中，因此本研究中白三叶配施商品有机肥显著提高了球囊霉属相对丰度[33−34]。

      本研究指出：白三叶和紫云英配施有机肥处理的孢子密度显著低于清耕组，可能与丛枝菌根真菌在土壤中的定殖率以及菌丝体密度受到肥料种类及投入水平的显著影响有关[35]。绿肥与施肥对球囊霉属的相对丰度存在显著交互作用，具体表现为在单施复合肥时，种植绿肥反而显著降低了球囊霉属相对丰度。复合肥提供的速效养分(尤其是磷)抑制了菌根依赖性强的球囊霉属的生长，同时绿肥与柑橘树体存在养分竞争，进一步削弱了共生关系[36]。而有机肥提供的稳定、缓释的养分环境缓解了养分抑制作用，白三叶作为优良宿主，其根系分泌物的代谢调控与球囊霉属存在明显的联系，并对植物产生全身效应[37−38]。

      • 3.2.   土壤化学性质是驱动丛枝菌根真菌群落构建的核心环境因子

    • 多项研究表明：环境因子对宿主共生丛枝菌根真菌菌根侵染程度、多样性、群落结构以及菌属丰度存在显著影响[39−41]。已有研究发现：增加土壤中氮可降低丛枝菌根真菌的菌根定殖率，从而影响丛枝菌根真菌群落结构[42]，而农田系统中氮肥的减施则可显著增加丛枝菌根真菌群落的丰度及多样性，进而促进植株根部丛枝菌根真菌的定殖[43]。因此，过高氮素可能直接抑制丛枝菌根真菌菌丝生长，或通过降低植物的菌根依赖性来间接影响群落组成。土壤pH可通过改变土壤养分离子有效性，影响土壤微生物细胞活动以及土壤胞外酶代谢进而影响微生物的生存与繁育，改变土壤微生物群落结构[44]。土壤pH是土壤中磷素有效性的主要影响因素，丛枝菌根真菌定殖率和孢子相对丰度随着土壤中可利用磷的增加而下降，随着pH的增加而上升[45]。TAWARAYA等[46]的研究发现：较低的土壤有效磷会促进丛枝菌根真菌的生长，同时丛枝菌根真菌群落丰富度和孢子密度也会显著提高，而随着土壤中有效磷的上升，部分丛枝菌根真菌的物种相对丰度下降[47]。不同的丛枝菌根真菌种类在土壤有机碳的利用和转化过程中具有不同的适应能力。这可能与丛枝菌根真菌类群在碳利用策略上存在生态位分异有关[48]。

      • 3.3.   绿肥配施有机肥通过优化丛枝菌根真菌群落提升其生态功能与稳定性

    • 本研究结果指出：与清耕处理相比，绿肥(尤其是鼠茅草、紫云英)处理和有机肥(商品有机肥、生物有机肥)处理的丛枝菌根真菌网络具有更高的模块化程度和更短的平均路径。更高的模块化意味着群落结构更紧凑，群落功能分区更为明确[49]。施肥可改变土壤微生物的共现网络拓扑结构，种植绿肥和有机肥处理下丛枝菌根真菌群落结构更加稳定，丛枝菌根真菌连接更紧密[50]。农业施肥方式可通过改变真菌群落多样性及网络模块化程度，并改变微生物功能而影响土壤肥力和作物产量[51]。在受到外部环境扰动时，丛枝菌根真菌群落的网络模块化程度增强，群落具有更高的稳定性和恢复力[52]。相关研究也指出：种植绿肥可显著增加土壤真菌群落的模块化程度，提高微生物群落的稳定性，提高土壤肥力和作物产量[53]。此外，有研究指出：真菌往往会更倾向于分解顽固性碳，绿肥和有机肥中的顽固性碳有利于土壤中真菌类群的生长，可增加真菌群落网络的复杂性[54]。施肥方式以及种植绿肥可通过优化丛枝菌根真菌群落进而提升群落功能。

      丛枝菌根真菌可通过植物根系介导微生物群落的形成，对真菌群落功能和植物功能可产生显著影响[55]。例如，丛枝菌根真菌和球囊素相关土壤蛋白的提升有助于土壤团聚体的形成，提高土壤团聚体稳定性；丛枝菌根真菌可通过扩大植物根部结构，刺激酸性磷酸酶分泌，增强土壤磷的活性与转运效率[56−57]。因此，农业系统需要建立健康的丛枝菌根真菌群落，改善作物营养状况，进而提升果实品质。丛枝菌根真菌与柑橘果实品质的关系不仅体现在其对土壤环境的改善，还通过直接影响柑橘的生理过程来提升果实品质。柑橘生长须通过光合作用来生成可溶性糖等营养有机物质，而接种丛枝菌根真菌可提高柑橘的光合作用，从而提高柑橘的固酸比和维生素C质量分数[58]。

    4.   结论

    • 绿肥与有机肥配施可显著改善柑橘果实品质与土壤肥力。生物有机肥配施鼠茅草或紫云英可显著提高果实维生素C质量分数和固酸比；同时提升土壤有机碳、全氮以及易提取态球囊霉素质量分数，优化土壤养分环境。绿肥种类与施肥方式共同调控丛枝菌根真菌群落组成与多样性，其中有机肥配施绿肥可显著提高丛枝菌根真菌孢子密度及群落模块化程度，其中鼠茅草与白三叶效果更佳，增强了丛枝菌根真菌群落的结构稳定性。土壤化学性质是驱动丛枝菌根真菌群落构建的关键因子，其中土壤全氮、有效磷、速效钾及pH与丛枝菌根真菌群落结构及孢子密度显著相关。有机肥通过调节土壤碳、氮、磷、钾平衡及pH，为丛枝菌根真菌提供适宜的生境，丛枝菌根真菌群落的优化，提升其生态功能并促进柑橘品质形成。综上所述，绿肥(以鼠茅草、白三叶为宜)配施生物有机肥可通过优化丛枝菌根真菌群落结构，提升土壤肥力与微生态稳定性，显著改善柑橘果实品质。建议在橘园生产中推广“绿肥+生物有机肥”替代部分化肥的施肥模式，为柑橘绿色可持续生产提供科学依据。

Reference (58)

Catalog

    浙ICP备11046845号

    Copyright by 2019 Editorial Department of Journal of Zhejiang A&F University

    Address: No. 666, Wu Su street, Ling'an District, Hangzhou, ZhejiangPost Code: 311300Tel: 0571-63732749

    E-Mail: :zlxb@zafu.edu.cn

    Supported by: Beijing Renhe Information Technology Co. Ltd  support: info@rhhz.net

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return