液态有机肥部分替代化肥对设施番茄根区细菌群落的影响

doi:10.13304/j.nykjdb.2021.0244

摘要/Abstract

摘要：

为研究液态有机肥部分替代化肥对设施番茄根区细菌群落的影响，以化学冲施肥为对照（T_CK），设置液态有机肥分别替代20%（T_R20）、30%（T_R30）和40%化学冲施肥（T_R40）3个处理，应用16S rRNA 高通量测序技术，分析番茄根区土壤样本中细菌的群落多样性、结构组成和差异。结果表明，在97%的相似水平下，共发现3 747个OTU，37个门，104个纲，268个目，441个科，730个属，1 401个种。3个处理和对照样品中共有OTU为1 338个，独有的OTU数值分别为140（T_R20）、129（T_R30）和263（T_R40）。T_R30处理的细菌丰富度和多样性最高，其优势菌门为变形菌门（Proteobacteria）、绿弯菌门（Chloroflexi）、酸杆菌门（Acidobacteria）、厚壁菌门（Firmicutes）；优势菌属为norank_c_subgroup_6、芽孢杆菌属（Bacillus）、norank_f_A4b。通过聚类分析、主成分分析、非度量多维尺度分析和共现性网络分析发现， T_R30和对照处理细菌的菌落组成相似度较高，共性最大。利用PICRUSt软件对细菌群落功能组成进行预测，发现各处理土壤样本的主要COG功能组成较为相似，但各COG相对丰度有所差异。综上所述，液态有机肥不同比例替代化学冲施肥能增加外源细菌种类，改变丰富度、多样性和菌落组成，以替代30%比例时效果最佳，为液态有机肥与化学冲施肥配施提供了理论依据。

关键词: 设施番茄, 液态有机肥, 化学冲施肥, 细菌, 群落结构

Abstract:

In order to study the effect of partial substitution of liquid organic fertilizer for chemical fertilizer on the bacterial community in the root zone of greenhouse tomato， chemical flushing fertilization was used as control （T_CK）， liquid organic fertilizer was set to replace 20% （T_R20）、30%（T_ R30） and 40% （T_R40） chemical flushing. 16S rRNA high-throughput sequencing technology was used to analyze the diversity， structural composition and differences of bacterial communities in different treatments. The results showed that， at 97% similarity level， 3 747 OTU， 37 phylas， 104 classes， 268 orders， 441 families， 730 genera and 1 401 species were found. There were shared 1 338 OUT in 4 treatments， and there were 140， 129 and 263 unique OTU values in T_R20， T_R30 and T_R40 treatments， respectively. The diversity of bacterial communities in T_R30 treatment was higher than other treatments， and the dominant phylums were Proteobacteria， Chloroflexi， Acidobacteria and Firmicutes， while the dominant genera were norank_c_subgroup_6、Bacillus and norank_f_A4b. Through cluster analysis， principal component analysis， non metric multi-dimensional scale analysis and co-occurrence network analysis， it was found that there were high similarity of bacterial composition between T_CK and T_R30 treatments. By PICRUSt software package， the functional composition of bacterial community showed that the main COGs were similar among four treatments， but the relative abundance of each COG changed. In conclusion， the different proportions of liquid organic fertilizer instead of chemical flushing fertilizer could increase the species of exogenous bacteria， change the richness， diversity and colony composition. And the effect of T_R30 treatment was the most excellent， which provided a theoretical basis for liquid organic fertilizer to replace chemical flushing fertilization.

Key words: greenhouse tomato, liquid organic fertilizer, inorganic fertilizer, bacterial, community structure

中图分类号:

S641.2

夏秀波, 李涛, 曹守军, 姚建刚, 王虹云, 张丽莉. 液态有机肥部分替代化肥对设施番茄根区细菌群落的影响[J]. 中国农业科技导报, 2022, 24(7): 187-196.

Xiubo XIA, Tao LI, Shoujun CAO, Jiangang YAO, Hongyun WANG, Lili ZHANG. Effect of Liquid Organic Fertilizer Partial Replacing Chemical Fertilizer on Bacterial Community in Greenhouse Tomato Root Zone[J]. Journal of Agricultural Science and Technology, 2022, 24(7): 187-196.

图/表 12

表１不同处理的施肥量

Table 1 Fertilization amount of different treatments

处理 Treatment	肥料类型 Fertilization type		次数 Times
处理 Treatment	化学冲施肥 Chemical fertilizer /（kg·hm^-2）	液态有机肥 Liquid organic fertilizer /（kg·L^-1）	次数 Times
T_CK	90.0	0	5
T_R20	72.0	15.0	5
T_R30	63.0	22.5	5
T_R40	54.0	30.0	5

表2 样本信息统计表

Table 2 Sample information statistics chart

样本 Sample	序列数 Sequence No.	碱基数 Base No.	平均序列长度 Mean sequence length/bp	最短序列长度 Min sequence length/bp	最长序列长度 Max sequence length/bp
T_CK	67 708	28 279 629	417.67	244	535
T_R20	57 661	23 977 066	415.83	258	519
T_R30	54 144	22 647 083	418.28	283	506
T_R40	72 046	30 248 429	419.85	336	461

图1 细菌的Sobs稀释曲线

Fig. 1 Rarefaction Sobs curves of OTU numbers

表3 样本细菌多样性指数

Table 3 Diversity index of Bacteria in samples

样品Sample	覆盖度 Coverage	Ace指数 Ace index	Chao指数 Chao index	Shannon指数 Shannon index	Simpson指数 Simpson index	Sobs指数 Sobs index
T_CK	0.991 6 a	3 002.15 b	3 003.27 b	6.52 a	0.005 7 c	2 694 a
T_R20	0.990 5 a	2 745.79 d	2 739.96 d	6.41 a	0.004 8 c	2 441 c
T_R30	0.989 5 a	3 081.20 a	3 064.07 a	6.46 a	0.006 8 b	2 721 a
T_R40	0.992 0 a	2 886.29 c	2 911.51 c	6.15 b	0.011 2 a	2 544 b

图2 细菌OUT分布水平上的Venn图

Fig.2 Venn map of the level of bacteria OTU distribution

图3 门水平各样本中细菌群落组成注：Others为样本中丰度占比均小于1%的进行合并。

Fig. 3 Composition of bacterial community on Phylum levelNote： Others are the samples with abundance ratio less than 1%.

图4 属水平各样品中细菌群落组成注：Others为样本中丰度占比均小于1%的进行合并。

Fig. 4 Composition of bacterial community on Genus levelNote： Others are the samples with abundance ratio less than 1%.

图5 不同样本细菌群落OTU水平层级聚类树系图

Fig. 5 Hierarchical clustering tree of bacteria on OTU level

图6 细菌群落OTU水平主成分分析

Fig. 6 Principal component analysis of bacteria on OTU level

图7 基于相似性>97%的OTU 水平上细菌非度量多维度分析

Fig. 7 NMDS plot of bacteria on OTU levelbased on similarity > 97%

图8 细菌群落在属水平的网络分析注：图中默认显示丰度大于50%的物种。

Fig. 8 Network analysis of bacteria on Genus levelNote： Species with abundance greater than 50% are shown by default.

图9 细菌群落的COG功能分类和相对丰度

Fig. 9 COG function classification and relative abundance of bacteria

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