氮肥减施对茶园土壤细菌群落结构的影响研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-1287

摘要/Abstract

摘要：

为了研究不同减氮模式对茶树地下部分细菌菌落结构的影响。设置16 kg/667 m²纯氮(减氮55.6%处理A),26 kg/667 m²纯氮(减氮27.8%处理B),36 kg/667 m²纯氮(常规施肥C),不施氮(CK)共4个施氮水平,对根际及非根际0-20 cm土壤、非根际20-40 cm土壤36个样品进行16S rRNA序列测定,利用IonS5^TMXL高通量测序平台对不同减氮处理的土壤细菌群落结构进行分析。试验结果表明:A、B、C三种施肥水平下,根际、非根际0-20 cm土壤及非根际20-40 cm土壤的细菌OTUs分别为505、854及835个,其中特异OTUs随着施氮量逐渐增加而降低。土壤细菌丰度和多样性分析显示施氮量为26 kg/667 m²(减氮处理B)在根际与非根际0-20 cm土壤中细菌丰度较高,细菌多样性最高,土壤碳通量的结果亦证明了减氮处理B土壤中微生物较多。施肥茶园地下部分在门水平上的优势菌基本为氮代谢相关菌类,其中共有3个优势类群,分别为变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、绿弯菌门(Chloroflexi)。在属的水平上,根际土壤与非根际土壤细菌优势菌完全不同。适当减氮处理B茶园土壤中细菌菌落多样性较多,丰度较高,并主要集中在养分吸收利用的根际及周边土壤中,有利于茶园养分的高效利用。

关键词: 茶, 细菌, 氮, 16S扩增测序, 根系, 土壤

Abstract:

This work is to study the effect of different nitrogen reduction modes on the bacterial community structure in the underground soil of tea plant. Four nitrogen levels were applied in the soil:16 kg/667 m²pure nitrogen(reducing nitrogen by 55.6%,treatment A),26 kg/667 m² pure nitrogen(reducing nitrogen by 27.8%,treatment B),36 kg/667 m² pure nitrogen(conventional fertilization,treatment C),and no nitrogen application(CK). The 36 samples of rhizosphere and non- rhizosphere soil of 0-20 cm and non- rhizosphere soil of 20-40 cm were sequenced by 16S rRNA,and the community structures of soil bacteria under different nitrogen reduction treatments were analyzed at IonS^5TMXL high-throughput sequencing platform. The results showed that the OTUs of the 0-20 cm rhizosphere and non rhizosphere soil and 20-40 cm non rhizosphere soil were 505,854 and 835 under the A,B and C treatment,respectively. The specific OTUs gradually decreased with the increase of nitrogen application. The bacterial abundance of the soil was higher in the rhizosphere soil and non- rhizosphere soil of 0-20 cm under nitrogen reduction treatment B,with the highest bacterial diversity. The results of soil carbon flux also confirmed that there were more microorganism in soil under nitrogen reduction treatment B. The dominant bacteria in the underground part of the fertilized tea garden at the phylum level were basically nitrogen metabolism related ones,among which there were three dominant groups,namely Proteobacteria,Actinobacteria and Chloroflexi. At the genus level,the dominant bacteria in the rhizosphere soil and non rhizosphere soil were totally different. The diversity of bacterial community in the tea garden soil was more and the abundance was higher under proper nitrogen reduction treatment,and it was mainly concentrated in the rhizosphere and surrounding soil of nutrient absorption and utilization,which is conducive to the efficient utilization of nutrients in tea garden.

Key words: tea, bacteria, nitrogen, 16S amplicon sequencing, root system, soil

向芬, 李维, 刘红艳, 银霞, 曾泽萱, 周凌云. 氮肥减施对茶园土壤细菌群落结构的影响研究[J]. 生物技术通报, 2021, 37(6): 49-57.

XIANG Fen, LI Wei, LIU Hong-yan, YIN Xia, ZENG Ze-xuan, ZHOU Ling-yun. Influence of Nitrogen Fertilizer Reduction on the Structure of Bacterial Community in Tea Garden Soil[J]. Biotechnology Bulletin, 2021, 37(6): 49-57.

图/表 11

表1 试验茶园施肥方式

Table 1 Fertilizer application methods in tea garden

处理 Treatment	施纯氮 Input of nitrogen/(kg·667 m^-2)	肥料种类 Fertilizer type	施用方式 Application method	施肥量 Fertilizer rate/(kg·667 m^-2)
CK	0	菜籽饼	基肥	0
		硫酸钾型复合肥	基肥	0
		尿素	追肥	0
减氮处理A	16	菜籽饼	基肥	9.25
		硫酸钾型复合肥	基肥	6.75
		尿素	追肥	0
减氮处理B	26	菜籽饼	基肥	9.25
		硫酸钾型复合肥	基肥	6.75
		尿素	追肥	10
常规施肥C	36	菜籽饼	基肥	9.25
		硫酸钾型复合肥	基肥	6.75
		尿素	追肥	20

图1 减氮处理对茶树叶面积指数的影响

Fig.1 Effect of nitrogen reduction on tea leaf area index

图2 不同土壤样品测序稀释曲线图 RCK,RA,RB,RC为根际土壤各施肥处理,CK20,A20,B20,C20为非根际0-20 cm土壤的各施肥处理,CK40,A40,B40,C40为非根际20-40 cm土壤的各施肥处理,下同

Fig.2 Rarefaction analysis of 16S libraries from different soil samples RCK, RA, RB, RC are fertilization treatments of rhizosphere soil; CK20, A20, B20, C20 are fertilization treatments of 0-20 cm non rhizosphere soil; CK40, A40, B40, C40 are fertilization treatments of 20-40 cm non rhizosphere soil. The same below

图3 不同土壤样品的Rank Abundance曲线

Fig.3 Rank Abundance curves of different soil samples

表2 各土壤样品微生物丰富度指数与多样性指数

Table 2 Diversity and richness index of bacteria from twelve samples

种类 Type	土层 Soil layer	施氮水平Nitrogen application level	观察物种 Observed species	Shannon指数 Shannon index	辛普森指数 Simpson index	Chao1指数 Chao1 index	Ace 指数 Ace index	PD进化树 PD whole tree
根际土壤		RCK	583.00±16.41	5.63±0.63	0.94±0.02	723.58±124.19	740.28±110.13	52.12±0.81
		RA	618.00±19.93	5.38±0.63	0.92±0.04	794.38±32.25	783.03±54.51	53.41±0.68
		RB	619.00±14.40	5.76±0.29	0.95±0.01	795.77±49.92	797.12±55.10	54.41±0.67
		RC	547.00±38.90	5.37±0.84	0.93±0.04	724.53±120.64	714.81±103.67	49.08±0.89
非根际土壤	0-20 cm	CK20	1186.00±153.96	8.24±0.84	0.99±0.01	1279.50±124.74	1301.71±139.38	93.92±4.44
		A20	957.00±115.66	7.39±0.41	0.98±0.01	1058.24±150.86	1085.27±159.78	82.99±4.40*
		B20	1069.00±83.16	7.72±0.11	0.99±0.01	1216.84±159.54	1247.27±149.99	88.46±5.92
		C20	1075.00±121.35	7.57±0.27	0.98±0.01	1241.87±170.12	1270.58±161.15	86.69±5.42
	20-40 cm	CK40	1110.00±110.59	7.52±0.30	0.98±0.01	1273.12±48.39	1286.25±53.62	91.49±3.11
		A40	893.00±18.08*	7.13±0.26	0.98±0.01	982.87±36.79*	1014.62±43.91*	74.89±0.72**
		B40	905.00±88.09*	7.04±0.37	0.97±0.02	993.23±156.54*	1029.39±179.62*	78.10±4.75*
		C40	1142.00±95.50	7.99±0.50*	0.99±0.01	1444.91±122.97	1440.32±187.43	102.06±9.56*

图4 不同土壤样品OTUs分布的Venn图 A, B, C分别表示根际土壤、非根际0-20 cm土壤,非根际20-40 cm土壤的Venn图

Fig.4 Venn diagrams of OTUs distribution of different soil samples A, B and C are Venn plots of rhizosphere soil, non rhizosphere 0-20 cm soil and non rhizosphere 20-40 cm soil

图5 门水平的细菌组成聚类堆积图

Fig.5 Cluster stacking graph of bacteria composition at phylum level

图6 属分类水平的细菌组成聚类堆积图

Fig.6 Cluster stacking graph of bacteria composition at genus level

图7 组间硝化螺旋菌门显著性差异统计图图中横轴为样品分组;纵向为对应物种的相对丰度。横线代表具有显著性差异的两个分组,没有则表示此物种在两个分组间不存在差异。“＊”表示两组间差异显著(P < 0.05),“＊＊”表示两组间差异极显著(P < 0.01)

Fig.7 Statistical chart of significant difference with Nitros-pirae between groups The horizontal axis in the figure is divided into sample groups. The relative abundance of the corresponding species was vertical. The horizontal line represents two groups with significant differences. * and ** indicate the difference between the two groups at 0.05, 0.01 level, respectively

图8 各土壤样品细菌群落主坐标分析

Fig.8 PCoA analysis of bacteria in different soil samples

图9 减氮处理对茶园土壤碳通量的影响

Fig.9 Effect of nitrogen reduction on soil carbon flux in tea garden

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