不同施肥处理对土壤微生物群落结构及西瓜枯萎病发生的影响

doi:10.16409/j.cnki.2095-039x.2018.04.014

摘要/Abstract

摘要： 研究不同肥料组合对西瓜枯萎病发生的影响，寻找与西瓜幼苗发病密切相关的菌群种类。设置3组不同土壤肥料组合，依次为对照组CK、T1、T2。利用Illumina Miseq高通量测序平台对试验组和对照组之间的土壤微生物群落组成在16S rDNA水平上进行测序分析。通过对测序结果在OTU和种群相对丰度的计算和分析发现，CK组中平均共获得28986条序列，T1组中平均共有30308条序列，T2组中平均共有29730个条序列；三组共有细菌OTU数量为860，占各组分平均总OTU数量的50%以上，其中芽胞杆菌属Bacillus、肠球菌属Enterococcus、乳球菌属Lactococcus、梭菌属Clostridium在T1组中相对丰度最高；而2种未知细菌unclassified_TM7-1（c）和unclassified_Ellin329（o）在T2组中的相对丰度值最高。西瓜枯萎病发病率情况为CK>T1>T2，施用棉隆+生物有机肥可以通过重建土壤生态环境，提高土壤细菌多样性，从而对西瓜枯萎病有显著的防治作用。

关键词: 西瓜, 土壤微生物群落, 生物有机肥, 16S rDNA, 枯萎病

Abstract: The objective of this study was to compare the effects of different fertilizer combinations on occurrence of fusarium wilt of watermelon and to excavate the closely related genus to the disease of watermelon seedlings. The composition of bacterial communities of soil samples fertilized by 3 different soil fertilizer combination groups, followed by the control group (CK), T1 and T2, were compared to that of bacterial communities by the Illumina Meseq platform targeting the V3-V4 regions of 16S rDNA. Through the sequencing results analysis in OTU and the relative abundance of the population, we have find 28986 tags in CK, 30308 tags in T1, and 29730 tags in T2, and there are 860 OTUs in common, which is more than 50% within average of each treatment. And in the genus top 10 bacteria population, the abundance of Bacillus, Enterococcus, Lactococcus, Clostridium in T1 was apparently higher than others, while the unclassified_TM7-1(c) and unclassified_Ellin329(o) were significantly high in T2. The incidence of watermelon Fusarium wilt is CK > T1 > T2. The application of Basamid+Bio-organic fertilizer can be used to rebuild the soil ecological environment and help to improve the soil bacterial diversity, which has a significant preventive effect on watermelon fasurium wilt.

Key words: watermelon, soil microbes, bio-organic fertilizer, 16S rDNA, Fusarium wilt

中图分类号:

S476
S651

朱菲莹, 田程, 张屹, 肖姬玲, 魏林, 梁志怀. 不同施肥处理对土壤微生物群落结构及西瓜枯萎病发生的影响[J]. 中国生物防治学报, 2018, 34(4): 589-597.

ZHU Feiying, TIAN Cheng, ZHANG Yi, XIAO Jiling, WEI Lin, LIANG Zhihuai. Effects of Different Fertilization Treatments on Soil Microbial Community Structure and the Occurrence of Watermelon Wilt[J]. journal1, 2018, 34(4): 589-597.

参考文献

[1] Ling N, Deng K, Song Y, et al. Variation of rhizospHere bacterial community in watermelon continuous mono-cropping soil by long-term application of a novel bioorganic fertilizer[J]. Microbiological Research, 2014, 169(7-8):570.
[2] 马利平, 高芬, 乔雄梧. 家畜沤肥浸渍液对黄瓜枯萎病的防治及作用机理探析[J]. 植物病理学报, 1999, 29(3):270-274.
[3] 蔡燕飞, 廖宗文, 章家恩. 生态有机肥对番茄青枯病及土壤微生物多样性的影响[J]. 应用生态学报, 2003, 14(3):349-353.
[4] 蔡燕飞, 廖宗文. FAME法分析施肥对番茄青枯病抑制和土壤健康恢复的效果[J]. 中国农业科学, 2003, 36(8):922-927.
[5] 张树生, 杨兴明, 黄启为, 等. 施用氨基酸肥料对连作条件下黄瓜的生物效应及土壤生物性状的影响[J]. 土壤学报, 2007, 44(4):689-694.
[6] 朱林, 彭宇, 袁飞, 等. 几种有机物对连作黄瓜生长的影响[J]. 安徽农业科学, 2001, 29(2):214-216.
[7] 徐福利, 梁银丽, 张成娥, 等. 日光温室土壤生物学特性与施肥的关系[J]. 水土保持研究, 2004, 11(1):20-22.
[8] 吕卫光, 张春兰, 袁飞. 有机肥减轻连作对黄瓜自毒作用的机制[J]. 上海农业学报, 2002, 18(2):52-56.
[9] 孙红霞, 武琴, 郑国祥, 等. EM对茄子、黄瓜抗连作障碍和增强土壤生物活性的效果[J]. 土壤, 2001(5):264-267.
[10] 张春兰, 吕卫光, 袁飞. 生物有机肥减轻设施栽培黄瓜连作障碍的效果[J]. 中国农学通报, 1999, 15(6):67-69.
[11] 邵丽, 谷洁, 张社奇, 等. 生物复混肥对土壤微生物功能多样性和土壤酶活性的影响[J]. 农业环境科学学报, 2012, 31(6):1153-1159.
[12] 邵丽, 谷洁, 张社奇, 等. 生物复混肥对土壤微生物群落功能多样性和微生物量的影响[J]. 中国生态农业学报, 2012, 20(6):746-751.
[13] Li W Z, Godzik A. Cd-hit:a fast program for clustering and comparing large sets of protein or nucleotide sequences[J]. Bioinformatics, 2006, 22(13):1658.
[14] Desantis T Z, Hugenholtz P, Larsen N, et al. Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB[J]. Applied and Environmental Microbiology, 2006, 72(7):5069-5072.
[15] Xiong W, Zhao Q, Zhao J, et al. Different continuous cropping spans significantly affect microbial community membership and structure in a vanilla-grown soil as revealed by deep pyrosequencing[J]. Microbial Ecology, 2015, 70(1):209.
[16] Zhou J, Wu L, Deng Y, et al. Reproducibility and quantitation of amplicon sequencing-based detection[J]. The ISME Journal, 2011, 5:1303.
[17] Chen L, Xun W, Sun L, et al. Effect of different long-term fertilization regimes on the viral community in an agricultural soil of Southern China[J]. European Journal of Soil Biology, 2014, 62(6):121-126.
[18] Lozupone C, Knight R. UniFrac:a new phylogenetic method for comparing microbial communities[J]. Applied and Environmental Microbiology, 2005, 71(12):8228.
[19] Xu W, Wang Z, Wu F. The effect of D123 wheat as a companion crop on soil enzyme activities, microbial biomass and microbial communities in the rhizosphere of watermelon[J]. Frontiers in Microbiology, 2015, 6:899.
[20] Schloss P D, Gevers D, Westcott S L. Reducing the effects of PCR amplification and sequencing artifacts on 16S rRNA-based studies[J]. PloS ONE, 2011, 6(12):e27310.
[21] Shiomi Y, Masaya N, Tomoko O, et al. Comparison of bacterial community structure in the rhizosphere of tomato plants grown in soils suppressive and conducive towards bacterial wilt[J]. Applied and Environental Microbiology, 1999, 65(9):3996-4001.
[22] Yang X. Effects of zero-tillage and applicationofmanure on soilmicrobial biomass C, N and P after sixteen years of cropping[J]. Acta Pedologica Sinica, 2002, 39(1):89-96.
[23] 杨元根, Campbell P. Biolog方法在区分城市土壤与农村土壤微生物特性上的应用[J]. 土壤学报, 2002, 39(4):582-589.
[24] Liu B, Cong T, Hu S, et al. Effect of organic sustainable, and conventional management stragies in grower field on soil physical, chemical, and biological factors and the incidence of southern blight[J]. Applied Soil Ecology, 2007, 37:202-214.
[25] Chen S N, Gu J, Gao H, et al. Effect of microbial fertilizer on microbial activity and microbial community diversity in the rhizosphere of wheat growing on the Loess Plateau[J]. Africa Journal of Microbiology Resource, 2011, 5(2):137-142.
[26] Zhou X, Wu F. Dynamics of the diversity of fungal and Fusarium communities during continuous cropping of cucumber in the greenhouse[J]. FEMS Microbiol Ecology, 2012, 80:469.
[27] 王媛. 棉隆和次氯酸钠土壤处理对设施连作西瓜生长发育及土壤微生物区系的影响[D]. 杨凌:西北农林科技大学, 2014.
[28] 张超, 卜东欣, 张鑫, 等. 棉隆对辣椒疫霉病的防效及对土壤微生物群落的影响[J]. 植物保护学报, 2016, 45(5):834-840.
[29] Huang L F, Song L X, Xia X J, et al. Plant-Soil feedbacks and soil sickness:from mechanisms to application in agriculture[J]. Journal of Chemical Ecology, 2013, 39(2):232-242.
[30] Li B, Zhang X, Guo F, et al. Characterization of tetracycline resistant bacterial community in saline activated sludge using batch stress incubation with high-throughput sequencing analysis[J]. Water Research, 2013, 47(13):4207.
[31] Li C G, Li X M, Kong W D, et al. Effect of monoculture soybean on soil microbial community in the Northeast China[J]. Plant and Soil, 2010, 330(1-2):423-433.
[32] Bokulich N A, Subramanian S, Faith J J, et al. Quality-filtering vastly improves diversity estimates from illumina amplicon sequencing[J]. Nature methods, 2013, 10(1):57-59.
[33] Philippot L, Raaijmakers J M, Lemanceau P, et al. Going back to the roots:the microbial ecology of the rhizosphere[J]. Nature reviews Microbiology, 2013, 11(11):789-99.
[34] Widmer F, Rasche F, Hartmann M, et a1. Community structures and substrate utilization of bacteria in soils from organic and conventional farming systems of the DOK long-term field experiment[J]. Applied Soil Ecology, 2006, 33(3):294-307.