不同温度烧制的生物炭对苹果连作土壤真菌群落结构的影响

doi:10.16420/j.issn.0513-353x.2022-0187

摘要/Abstract

摘要：

根皮苷类酚酸类物质是造成苹果连作障碍的重要因素之一。以废弃苹果枝条为原料，研究不同温度烧制的生物炭对根皮苷胁迫下平邑甜茶（苹果砧木）连作土盆栽幼苗土壤中真菌群落结构及其多样性的影响。试验设置8个处理，分别为：连作土（对照）、连作土壤中分别添加50 mg · kg^-1根皮苷、300 ℃生物炭、400 ℃生物炭、500 ℃生物炭、300 ℃生物炭 + 根皮苷、400 ℃生物炭 + 根皮苷、500 ℃生物炭 + 根皮苷，盆栽处理120 d后，运用高通量测序技术对土壤真菌群落结构进行分析。结果表明：添加生物炭可以促进根皮苷胁迫下平邑甜茶幼苗的生长，改变连作土壤的真菌群落结构，使土壤真菌操作分类单元（operational taxonomic unit，OUT）特有数增加；通过PCoA分析以及UPGMA聚类分析得出，添加生物炭后土壤真菌的丰富度和多样性显著提高；生物炭对不同分类水平上的土壤真菌结构和功能产生了一定影响，土壤优势菌丰度变化较大。生物炭可通过改善土壤真菌群落结构缓解苹果连作障碍。

关键词: 苹果, 连作障碍, 根皮苷, 生物炭, 高通量测序, 土壤真菌群落

Abstract:

Phenolic acids，such as phlorizin，are one of the important factors that cause apple replant disease. In this study，the effects of biochar with different firing temperatures on the fungi community structure and diversity of apple（Malus hupehensis）replanted soil were studied by pot experiments of apple seedlings in replanted soil. There are eight treatments，which are replant soil（control），and addition of 300 ℃ biochar，400 ℃ biochar，500 ℃ biochar，50 mg · kg^-1 phlorizin，300 ℃ biochar + phlorizin，400 ℃ biochar + phlorizin，500 ℃ biochar + phlorizin. After 120 days of pot experiment，the soil fungal communities were determined by high-throughput sequencing technology. The results showed that added biochar could improve the growth of apple seedlings under phlorizin stress，and change the fungal community structure of continuous cropping soil and increase the unique number of soil fungi OUT（operational taxonomic unit）. PCoA analysis and UPGMA cluster analysis showed that soil fungal community structure and diversity with the addition of biochar were significantly different from the control soil. The addition of biochar had a certain impact on the structure and function of soil fungi at different species levels，making the abundance of dominant soil fungi change greatly. The addition of biochar had a certain effect on alleviating continuous cropping obstacles by improving the structure of fungal communities.

Key words: apple, replant disease, phlorizin, biochar, high-throughput sequencing, soil fungi community

刘英浩, 李允, 吕伟静, 陈冉, 姜伟涛, 尹承苗, 毛志泉, 王艳芳. 不同温度烧制的生物炭对苹果连作土壤真菌群落结构的影响[J]. 园艺学报, 2024, 51(8): 1853-1867.

LIU Yinghao, LI Yun, LÜ Weijing, CHEN Ran, JIANG Weitao, YIN Chengmiao, MAO Zhiquan, WANG Yanfang. Effects of Different Temperatures’ Biochar on Soil Fungi Community Structure Under Apple Replantation[J]. Acta Horticulturae Sinica, 2024, 51(8): 1853-1867.

图/表 9

表1 苹果连作土壤添加不同温度烧制的生物炭和根皮苷对真菌α多样性指数的影响

Table 1 Effect of different firing temperature of biochar and phlorizin on fungal diversity index in apple continuous cropping soil

代号 Code	处理 Treatment	Sobs指数 Sobs index	Chao 1指数 Chao 1 index	Ace指数 Ace index	Shannon指数 Shannon index
CK	连作土空白对照Blank control of replanted soil	585.00 ± 22.54 b	641.73 ± 5.73 d	642.32 ± 7.02 cd	3.49 ± 0.12 ab
P	根皮苷Phloridzin	681.00 ± 26.21 a	747.70 ± 16.67 abc	750.84 ± 16.66 ab	3.77 ± 0.03 a
300B	300 ℃生物炭Biochar made at 300 ℃	590.33 ± 48.35 b	669.17 ± 40.70 cd	654.83 ± 36.94 c	3.09 ± 0.26 ab
400B	400 ℃生物炭Biochar made at 400 ℃	729.33 ± 30.24 a	829.80 ± 29.00 a	832.68 ± 23.90 a	3.26 ± 0.27 ab
500B	500 ℃生物炭Biochar made at 500 ℃	669.67 ± 36.30 a	776.72 ± 26.87 ab	785.27 ± 26.59 a	2.97 ± 0.12 bc
300BP	300 ℃生物炭 + 根皮苷Biochar made at 300 ℃ + Phloridzin	571.00 ± 18.36 b	683.55 ± 11.63 bcd	683.32 ± 11.00 bcd	2.37 ± 0.10 c
400BP	400 ℃生物炭 + 根皮苷Biochar made at 400 ℃ + Phloridzin	728.00 ± 87.73 a	814.93 ± 57.79 a	799.30 ± 54.66 a	3.78 ± 0.07 a
500BP	500 ℃生物炭 + 根皮苷Biochar made at 500 ℃ + Phloridzin	655.33 ± 20.11 a	745.44 ± 6.70 abc	743.63 ± 3.27 ab	3.57 ± 0.40 ab

图1 不同温度烧制的生物炭处理苹果连作土壤的真菌OUT数量韦恩图各处理名称及代号详见表1。下同。

Fig. 1 The Venn diagram of fungal OUT number in apple continuous cropping soil treated with biochar calcined at different temperatures Each treatment name and code is detailed in Table 1. The same below.

图2 不同处理的真菌PCoA分析图

Fig. 2 PCoA analysis chart of fungi with different treatments

图3 不同处理的真菌Weighted Unifrac指数热图

Fig. 3 Heatmap of Weighted Unifrac index of Fungi with different treatments

图4 不同处理的真菌UPGMA聚类树

Fig. 4 Fungus UPGMA clustering tree of different treatments

图5 不同处理真菌门水平菌群分布

Fig. 5 Distribution of fungi phylum bacteria in different treatments

图6 不同处理LEfSe分析环状树图圆圈直径与相对丰度呈正比，不同颜色节点代表该颜色分组起重要影响的微生物类群。

Fig. 6 Different treatments LEfSe analysis ring tree diagram The diameter of the circle is proportional to the relative abundance，and the different color nodes represent the microbial groups that play an important role in the color group.

图7 不同处理LEfSe分析柱状图图中仅显示LDA的绝对值大于2的物种。

Fig. 7 LEfSe analysis histogram of different treatments It shows only species with absolute value of LDA greater than 2.

图8 不同温度烧制的生物炭对平邑甜茶幼苗株高和植株干质量的影响

Fig. 8 Effect of different firing temperature of biochar on Malus hupehensis seedling plant height and dry weight

参考文献 52

[1]	Agegnehu G A, Srivastava B A K, Michael I, Bird A. 2017. The role of biochar and biochar-compost in improving soil quality and crop performance:a review. Applied Soil Ecology,119:156-170.
[2]	Al-Wabel M I, Al-Omran A, El-Naggar Ahmed H, Nadeem M, Usman Adel R A. 2013. Pyrolysis temperature induced changes in characteristics and chemical composition of biochar produced from conocarpus wastes. Bioresource Technology,131:374-379.
[3]	Ameloot N, De N S, Jegajeevagan K, Yildiz G, Buchan D, Funkuin Y N, Prins W, Bouckaert L, Sleutel S. 2013. Short-term CO₂ and N₂O emissions and microbial properties of biochar amended sandy loam soils. Soil Biology and Biochemistry,57:401-410.
[4]	Bao L M, Liu Y Y, Ding Y F, Shang J J, Wei Y L, Tan Y, Zi F T. 2022. Interactions between phenolic acids and microorganisms in rhizospheric soil from continuous cropping of Panax notoginseng. Frontiers in Microbiology,13:791603.
[5]	Barbi F, Prudent E, Vallon L, Buée M, Dubost A, Legout A, Marmeisse R, Fraissinet-Tachet L, Luis P. 2016. Tree species select diverse soil fungal communities expressing different sets of lignocellulolytic enzyme-encoding genes. Soil Biology and Biochemistry,100:149-159.
[6]	Bouskill N J, Lim H C, Borglin S, Salve R, Wood T E, Silver W L, Brodie E L. 2013. Pre-exposure to drought increases the resistance of tropical forest soil bacterial communities to extended drought. The ISME Journal, 7 (2):384-394.
[7]	Chen H M, Ma J Y, Wei J X, Gong X, Yu X C, Guo H, Zhao Y W. 2018. Biochar increases plant growth and alters microbial communities via regulating the moisture and temperature of green roof substrates. Science of the Total Environment,635:333-342.
[8]	Cheng J R, Zhao Y H, Li J Y, Jiang H X, Liu P, Yang T, Bao Z Z, Zhou B Q, Zhou X Y, Liu X L. 2014. Identification and evaluation of a potential biocontrol agent,Bacillus subtilis,against Fusarium sp. in apple seedlings. Annals of Microbiology, 64 (1):377-383.
[9]	Cui Y X, Bing H J, Fang L C, Wu Y H, Yu J L, Shen G T, Jiang M, Wang X, Zhang X C. 2019. Diversity patterns of the rhizosphere and bulk soil microbial communities along an altitudinal gradient in an alpine ecosystem of the eastern Tibetan Plateau. Geoderma,338:118-127.
[10]	Duan C X, Qin Z H, Yang Z H, Li W X, Sun S L, Zhu Z D, Wang X M. 2016. Identification of pathogenic Fusarium spp. causing maize ear rot and potential mycotoxin production in China. Toxins, 8 (6):186.
[11]	Franke-Whittle I H, Manici L M, Insam H, Stres B. 2015. Rhizosphere bacteria and fungi associated with plant growth in soils of three replanted apple orchards. Plant and Soil, 395 (1):317-333.
[12]	Gregory S J, Anderson C W N, Camps A M, McManus M T. 2014. Response of plant and soil microbes to biochar amendment of an arsenic-contaminated soil. Agriculture,Ecosystems & Environment,191:133-141.
[13]	Gul S, Whalen J K, Thomas B W, Sachdeva V, Deng H Y. 2015. Physico-chemical properties and microbial responses in biochar-amended soils:mechanisms and future directions. Agriculture,Ecosystems & Environment,206:46-59.
[14]	Hu L A, Cao L X, Zhang R D. 2014. Bacterial and fungal taxon changes in soil microbial community composition induced by short-term biochar amendment in red oxidized loam soil. World Journal of Microbiology and Biotechnology, 30 (3):1085-1092.
[15]	Ju R C, Zhao Y H, Li J Y, Jiang H X, Liu P. 2014. Identification and evaluation of a potential biocontrol agent,Bacillus subtilis,against Fusarium sp. in apple seedlings. Annals of Microbiology, 64 (1):377-383.
[16]	Lan Jianglin, Xiao Rongfeng, Liu Bo, Zhu Yujing, Che Jianmei, Lin Kangmei. 2012. Growth kinetic model of Fusarium oxysporum under different pH. Chinese Journal of Eco-Agriculture, 20 (12):1532-1538. (in Chinese)
	蓝江林, 肖荣凤, 刘波, 朱育菁, 车建美, 林抗美. 2012. pH胁迫下尖孢镰刀菌生长动力学模型. 中国生态农业学报, 20 (11):1532-1538.
[17]	Lauber C L, Strickland M S, Bradford M A, Fierer N. 2008. The influence of soil properties on the structure of bacterial and fungal communities across land-use types. Soil Biology and Biochemistry, 40 (9):2407-2415.
[18]	Ma Z T, Du W Y, Yan Z B, Chen X S, Wang Y F, Mao Z Q. 2020. Removal of phloridzin by chitosan-modified biochar prepared from apple branches. Analytical Letters, 54 (5):903-918.
[19]	Ma Zhiting. 2020. Adsorption mechanism and application of biochar and chitosan modified biochar adsorbing phloridzin[M. D. Dissertation]. Tai’an: Shandong Agricultural University. (in Chinese)
	马志婷. 2020. 生物炭及壳聚糖改性生物炭吸附根皮苷机理及其应用[硕士论文]. 泰安: 山东农业大学.
[20]	Mazzola M, Manici L M. 2012. Apple replant disease:role of microbial ecology in cause and control. Annual Review of Phytopathology,50:45-65.
[21]	Mazzola M, Reardon Catherine L, Brown J. 2012. Initial pythium species composition and brassicaceae seed meal type influence extent of Pythium-induced plant growth suppression in soil. Soil Biology & Biochemistry,48:20-27.
[22]	Oleszczuk P, Jośko I, Futa B, Pasieczna-Patkowska Sylwia, Pałys Edward, Kraska Piotr. 2014. Effect of pesticides on microorganisms,enzymatic activity and plant in biochar-amended soil. Geoderma,214-215:10-18.
[23]	Prasanna R, Kanchan A, Kaur S, Ramakrishnan B, Ranjan K, Singh M C, Hasan M, Saxena A K, Shivay Y S. 2016. Chrysanthemum growth gains from beneficial microbial interactions and fertility improvements in soil under protected cultivation. Horticultural Plant Journal, 2 (4):229-239.
[24]	Pruesse E, Quast C, Knittel K, Fuchs B M, Ludwig W, Peplies J, Glöckner F O. 2007. Silva:a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Research, 35 (21):7188-7196. doi: 10.1093/nar/gkm864 pmid: 17947321
[25]	Quilliam R S, Glanville H C, Wade S C, Jones D L. 2013. Life in the‘charosphere’-does biochar in agricultural soil provide a significant habitat for microorganisms? Soil Biology and Biochemistry,65:287-293.
[26]	Randolph P, Bansode R R, Hassan O A, Rehrah D J, Ravella R, Reddy M R, Watts D W, Novak J M, Ahmedna M. 2017. Effect of biochars produced from solid organic municipal waste on soil quality parameters. Journal of Environmental Management,192:271-280.
[27]	Sewell G W F. 2008. Effect of Pythium species on the growth of apple and their possible causal role in apple replant disease. Annals of Applied Biology, 97 (1):31-42.
[28]	Song W P, Guo M X. 2012. Quality variations of poultry litter biochar generated at different pyrolysis temperatures. Journal of Analytical and Applied Pyrolysis,94:138-145.
[29]	Spanic V, Katanic Z, Sulyok M, Krska R, Arkanj B. 2020. Multiple fungal metabolites including mycotoxins in naturally infected and Fusarium-inoculated wheat samples. Microorganisms,8:578.
[30]	Sun Bulei, Wang Yanfang, Zhang Xianfu, Shen Xiang, Chen Xuesen, Mao Zhiquan. 2015. Effect of residual roots on the biomass of Malus hupehensis seedlings,phenolic acids and microbiology in the soil of continuous cropping. Acta Horticulturae Sinica, 42 (1):131-139. (in Chinese)
	孙步蕾, 王艳芳, 张先富, 沈向, 陈学森, 毛志泉. 2015. 连作土壤中残根对平邑甜茶幼苗生物量及土壤酚酸类物质和微生物的影响. 园艺学报, 42 (1):131-139.
[31]	Tewoldemedhin Y T, Mazzola M, Labuschagne I, McLeod A. 2011. A multi-phasic approach reveals that apple replant disease is caused by multiple biological agents,with some agents acting synergistically. Soil Biology & Biochemistry, 43 (9):1917-1927.
[32]	Tian Xue, Zhou Wenjun, Zheng Weiguo, Gao Yuhui, Cao Huaying. 2021. Study on adsorption and desorption of nitrogen and phosphorus by biochar prepared from garden waste branches and leaves at different temperatures. Acta Agriculturae Jiangxi, 33 (1):98-104. (in Chinese)
	田雪, 周文君, 郑卫国, 高育慧, 曹华英. 2021. 不同温度制备的园林废弃物生物炭对氮磷吸附解吸的研究. 江西农业学报, 33 (1):98-104.
[33]	Toju H, Tanabe A S, Yamamoto S, Sato H. 2012. High-coverage ITS primers for the DNA-based identification of ascomycetes and basidiomycetes in environmental samples. PLoS ONE, 7 (7):e40863.
[34]	Treseder K K. 2013. The extent of mycorrhizal colonization of roots and its influence on plant growth and phosphorus content. Plant and Soil, 371 (1):1-13.
[35]	Wang G S, Yin C M, Pan F B, Wang X B, Xiang L, Wang Y F, Wang J Z, Tian C P, Chen J, Mao Z Q. 2018. Analysis of the fungal community in apple replanted soil around Bohai gulf. Horticultural Plant Journal, 4 (5):175-181.
[36]	Wang Q, Garrity G M, Tiedje J M, Cole J R. 2007. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Applied and Environmental Microbiology, 73 (16):5261-5267. doi: 10.1128/AEM.00062-07 pmid: 17586664
[37]	Wang Qingqing, Hu Yanli, Zhou Hui, Zhan Xing, Mao Zhiquan, Zhu Shuhua. 2012. Effects of phloridzin on the tricarboxylic acid cycle enzymes of roots of Malus hupehensis Rehd. Scientia Agricultura Sinica, 45 (15):3108-3114. (in Chinese) doi: 10.3864/j.issn.0578-1752.2012.15.012
	王青青, 胡艳丽, 周慧, 展星, 毛志泉, 朱树华. 2012. 根皮苷对平邑甜茶根系TCA循环酶的影响. 中国农业科学, 45 (15):3108-3114. doi: 10.3864/j.issn.0578-1752.2012.15.012
[38]	Wang Xiaobao, Wang Gongshuai, Liu Yusong, Chen Xuesen, Shen Xiang, Yin Chengmiao, Mao Zhiquan. 2018. Correlation analysis of apple replant disease and soil fungal community structure in the northwest loess plateau area. Acta Horticulturae Sinica, 45 (5):855-864. (in Chinese) doi: 10.16420/j.issn.0513-353x.2017-0668
	王晓宝, 王功帅, 刘宇松, 陈学森, 沈向, 尹承苗, 毛志泉. 2018. 西北黄土高原地区苹果连作障碍与土壤真菌群落结构的相关性分析. 园艺学报, 45 (5):855-864. doi: 10.16420/j.issn.0513-353x.2017-0668
[39]	Wang Xiaoqi, Jiang Weitao, Yao Yuanyuan, Yin Chengmiao, Chen Xuesen, Mao Zhiquan. 2020. Research progress on soil microorganisms that hinder continuous apple cropping. Acta Horticulturae Sinica, 47 (11):2223-2237. (in Chinese)
	王晓琪, 姜伟涛, 姚媛媛, 尹承苗, 陈学森, 毛志泉. 2020. 苹果连作障碍土壤微生物的研究进展. 园艺学报, 47 (11):2223-2237.
[40]	Wang Y F, Ma Z T, Wang X W, Sun Q R, Dong H Q, Wang G S, Chen X S, Yin C M, Han Z H, Mao Z Q. 2019. Effects of biochar on the growth of apple seedlings,soil enzyme activities and fungal communities in replant disease soil. Scientia Horticulturae,256:108641.
[41]	Wang Yanfang, Pan Fengbing, Zhan Xing, Wang Gongshuai, Zhang Guodong, Hu Yanli, Chen Xuesen, Mao Zhiquan. 2015. Effects of five kinds of phenolic acid on the function of mitochondria and antioxidant systems in roots of Malus hupehensis Rehd. seedlings. Acta Ecologica Sinica, 35 (19):6566-6573. (in Chinese)
	王艳芳, 潘凤兵, 展星, 王功帅, 张国栋, 胡艳丽, 陈学森, 毛志泉. 2015. 连作苹果土壤酚酸对平邑甜茶幼苗的影响. 生态学报, 35 (19):6566-6573.
[42]	Wang Yanfang, Xiang Li, Xu Shaozhuo, Wang Sen, Wang Xiaowei, Chen Xuesen, Mao Zhiquan, Zhang Min. 2017. Effects of biochar and chitin combined application on Malus hupehensis Rehd. seedlings and soil environment under replanting conditions. Scientia Agricultura Sinica, 50 (4):711-719. (in Chinese)
	王艳芳, 相立, 徐少卓, 王森, 王晓伟, 陈学森, 毛志泉, 张民. 2017. 生物炭与甲壳素配施对连作平邑甜茶幼苗及土壤环境的影响. 中国农业科学, 50 (4):711-719. doi: 10.3864/j.issn.0578-1752.2017.04.011
[43]	Xiang L, Wang M, Pan F B, Wang G S, Jiang W T, Wang Y F, Chen X S, Yin C M, Mao Z Q. 2021. Transcriptome analysis Malus domestica ‘M9T337’root molecular responses to Fusarium solani infection. Physiological and Molecular Plant Pathology,113:101567.
[44]	Xu L H, Ravnskov S, Larsen J, Nilsson R H, Nicolaisen M. 2012. Soil fungal community structure along a soil health gradient in pea fields examined using deep amplicon sequencing. Soil Biology and Biochemistry,46:26-32.
[45]	Xu Liang, Yu Xiaona, Li Xueli, Wang Yuelin, Song Jiaqian, Ye Xiefeng, Lu Jian. 2021. Physico-chemical characteristics of biochars prepared by pyrolysis of tobacco-stalk under different temperatures. Chinese Journal of Soil Science, 52 (1):75-81. (in Chinese)
	徐亮, 于晓娜, 李雪利, 王悦霖, 宋佳倩, 叶协锋, 卢剑. 2021. 不同热解温度制备的烟秆生物炭理化特征分析. 土壤通报, 52 (1):75-81.
[46]	Yin C M, Xiang L, Wang G S, Wang Y F, Shen X, Chen X S, Mao Z Q. 2016. How to plant apple trees to reduce replant disease in apple orchard:a study on the phenolic acid of the replanted apple orchard. PLoS ONE,11:e0167347.
[47]	Yin C M, Xiang L, Wang G S, Wang Y F, Shen X, Chen X S, Mao Z Q. 2017. Phloridzin promotes the growth of Fusarium moniliforme(Fusarium verticillioides). Scientia Horticulturae,214:187-194.
[48]	Yin Chengmiao, Wang Mei, Wang Jiayan, Chen Xuesen, Shen Xiang, Zhang Min, Mao Zhiquan. 2017. The research advance on apple replant disease. Acta Horticulturae Sinica, 44 (11):2215-2230. (in Chinese) doi: 10.16420/j.issn.0513-353x.2017-0524
	尹承苗, 王玫, 王嘉艳, 陈学森, 沈向, 张民, 毛志泉. 2017. 苹果连作障碍研究进展. 园艺学报, 44 (11):2215-2230.
[49]	Yuan J H, Xu R K, Zhang H. 2011. The forms of alkalis in the biochar produced from crop residues at different temperatures. Bioresource Technology, 102 (3):3488-3497.
[50]	Zhang P, Sun H W, Min L J, Ren C. 2018. Biochars change the sorption and degradation of thiacloprid in soil:insights into chemical and biological mechanisms. Environmental Pollution,236:158-167.
[51]	Zhang Yaobo, Mao Zhiquan, Zhu Shuhua. 2011. Effects of phenolic acids on mitochondria and the activity of antioxidant enzymes in roots of seedlings of Malus hupehensis Rehd. Scientia Agricultura Sinica, 44 (15):3177-3184. (in Chinese)
	张兆波, 毛志泉, 朱树华. 2011. 6种酚酸类物质对平邑甜茶幼苗根系线粒体及抗氧化酶活性的影响. 中国农业科学, 44 (15):3177-3184. doi: 10.3864/j.issn.0578-1752.2011.15.013
[52]	Zhu Qilin, Suo Long, Liu Lijun, Zhang Xuebin, Liu Jinxia, Meng Lei, He Qiuxiang, Ke Yongchun. 2022. Effect of pyrolysis temperature on the physicochemical properties of biochar from different materials in Hainan. Journal of Tropical Crops, 43 (1):216-223. (in Chinese)
	朱启林, 索龙, 刘丽君, 张雪彬, 刘金霞, 孟磊, 何秋香, 柯用春. 2022. 裂解温度对海南不同材料生物炭理化特性的影响. 热带作物学报, 43 (1):216-223. doi: 10.3969/j.issn.1000-2561.2022.01.026