朱杰,
刘海,
吴邦魁,
袁峰,
刘章勇,
金涛^,
长江大学湿地生态与农业利用教育部工程研究中心/湖北省涝渍灾害与湿地农业重点实验室 荆州 434025
基金项目: 国家重点研发计划项目2017YFD0800102

详细信息

作者简介:朱杰, 主要研究方向为土壤微生物分子生态。E-mail:hbzj0806@163.com

通讯作者:金涛, 主要研究方向为土壤碳氮循环与生态环境。E-mail:jintao165@126.com

中图分类号:S154.3

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出版历程

收稿日期:2017-12-14
录用日期:2018-05-04
刊出日期:2018-09-01

Effects of integrated rice-crayfish farming system on community structure and diversity of nirK denitrification microbe in paddy soils

ZHU Jie,
LIU Hai,
WU Bangkui,
YUAN Feng,
LIU Zhangyong,
JIN Tao^,
Yangtze University, Engineering Research Center of Ecology and Agricultural Use of Wetland, Ministry of Education/Hubei Key Laboratory of Waterlogging Disaster and Wetland Agriculture, Jingzhou 434025, China
Funds: the National Key Research and Development Program of China2017YFD0800102

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Corresponding author:JIN Tao, E-mail:jintao165@126.com

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摘要
摘要:稻虾共作是水稻种植与克氏螯虾共作形成的互利共生的稻田种养复合生态模式。目前对稻虾共作模式稻田反硝化微生物多样性和群落结构的影响尚不清楚。本研究以江汉平原常规中稻模式（MR）为对照，设置连续3年（2014-2016年）稻虾共作模式（CR）为处理，通过特异引物提取中稻抽穗期稻田土壤nirK基因，采用Illumina Miseq高通量测序技术，探讨稻虾共作模式对稻田土壤nirK反硝化微生物多样性和群落结构的影响。结果表明：稻虾共作模式显著提升水稻抽穗期稻田土壤中硝态氮、全氮及全碳的含量，对土壤碳氮比、碱解氮和铵态氮含量没有显著影响。稻虾共作模式显著增加稻田土壤nirK基因微生物的丰富度指数，但对nirK基因微生物的多样性指数影响不显著。稻虾共作模式改变了nirK基因微生物在目、科、属、种水平的群落组成，较常规中稻模式，稻虾共作模式在各分类水平组成类群均减少；稻虾共作模式较常规中稻模式改变了目的种类，对共有目相对丰度没有显著性改变。RDA分析表明稻虾共作模式对土壤nirK基因菌群的群落结构有一定的改变，但稻虾共作模式与常规中稻模式在群落结构上仍保留着一定的相似性。硝态氮含量是影响nirK反硝化细菌群落结构的主效因子。可见，稻虾共作模式对微生物多样性指数没有显著影响，但显著增加了微生物丰富度指数，改变了稻田土壤nirK反硝化微生物在目、科、属、种的群落结构。
关键词:江汉平原/
稻虾共作/
克氏螯虾/
nirK基因/
反硝化微生物/
群落结构/
硝态氮/
高通量测序技术
Abstract:Integrated rice-crayfish farming system is a symbiotic ecological model applicable in paddy field cultivation that is based on the combination of rice planting and clawed crayfish breeding in waterlogged conditions. In spite of so many efforts, the effects of integrated rice-crayfish farming system on denitrifying micro-organism diversity and community structure have remained unclear. In this study, we analyzed soil samples from both consecutive treatment of integrated rice-crayfish farming system (CR) in 2014-2016 and traditional paddy field (MR) treatment in order to investigate the effects of integrated rice-crayfish farming system on microbial diversity and community structure of nirK denitrification in paddy soils.. This was done by extracting soil nirK gene from rice field at heading stage using specific primers and Illumina Miseq high-throughput sequencing technology. The results showed that CR significantly increased the contents of nitrate nitrogen, total nitrogen and total carbon in paddy soils at heading stage, but had no significant effect on the ratio of carbon to nitrogen, contents of available nitrogen and ammonium nitrogen in soil. Compared with MR, CR significantly increased nirK gene abundance in soil, but did not significantly change its diversity. CR treatment changed the composition of nirK gene micro-organisms in the levels of order, family, genus and species. Compared with MR, CR reduced all taxonomic groups. The analysis of relative abundance of order showed no significant difference between CR and MR treatments. CR treatment changed species order, but did not change the relative abundance of common orders. RDA analysis showed that CR significantly changed community structure of nirK gene in soil. Nitrate nitrogen content was the main factor affecting the community structure of nirK denitrifying bacteria. It was obvious that rice-crayfish farming system had no significant effect on microbial diversity, but significantly increased microbial abundance index. In addition, it changed nirK denitrifying microbial community structure in terms of order, family, genus and species.
Key words:Jianghan Plain/
Rice-crayfish farming system/
Clawed crayfish/
nirK gene/
Denitrification microbe/
Community structure/
Nitrate nitrogen/
High-through put sequencing technology

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图1稻虾共作模式(CR)和常规中稻模式(MR)土壤样品OTU水平Coverage指数稀释性曲线
Figure1.Rarefaction curves of OTU level coverage indexes in soil samples of integrated rice-crayfish farming system (CR) and conventional middle rice system (MR)


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图2稻虾共作模式(CR)和常规中稻模式(MR)土壤样品nirK基因微生物物种分类学组成Venn图
a、b、c、d分别表示目、科、属、种4种分类学水平。
Figure2.Venn drawings of nirK gene species taxonomy of integrated rice-crayfish farming system (CR) and conventional middle rice system (MR)
a, b, c and d respectively show the taxonomic levels of order, family, genus and species.


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图3稻虾共作模式(CR)和常规中稻模式(MR)样品nirK基因目水平组成
Halobacteriales:盐细菌; Burkholderiales:伯克霍尔德氏菌; unclassified_c_Betaproteobacteria:未分类c-β蛋白酶细菌; unclassified_p_proteobacteria:未分类p-蛋白细菌; Rhizobiales:根瘤菌; unclassified_c__Alphaproteobacteria:未分类_c__α-蛋白细菌; Nitrosomonadales:亚硝化单胞菌目; Rhodospirillales:红螺菌目。
Figure3.Order distributions of nirK gene in different soil samples of integrated rice-crayfish farming system (CR) and conventional middle rice system (MR)


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图4稻虾共作模式和常规中稻模式土壤化学性质与细菌群落(属水平)的冗余分析
MR1、MR2和MR3分别表示常规中稻模式的3次重复, CR1、CR2和CR3分别表示稻虾共作模式的3次重复。TC:土壤总碳; NO3N:硝态氮; NH4N:铵态氮; PH: pH; AN:碱解氮。
Figure4.Results from redundancy analysis to explore the relationship between soil bacterial community (at genus level) and soil physiochemical characteristics of integrated rice-crayfish farming system and conventional mid-rice model
MR1, MR2 and MR3 mean three replicates of the conventional middle rice; CR1, CR2 and CR3 mean three replicates of the rice-crayfish culture. TC: soil total carbon; NO3N: nitrate nitrogen; NH4N: ammonium nitrogen; PH: pH; AN: available nitrogen.


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表1稻虾共作模式对稻田土壤理化性质的影响
Table1.Effects of integrated rice-crayfish farming on soil physicochemical properties in paddy field

种植模式 Planting pattern	pH	碱解氮 Available nitrogen (mg·kg-1)	硝态氮 NO3--N (mg·kg-1)	铵态氮 NH4+-N (mg·kg-1)	全碳 Total carbon (g·kg-1)	全氮 Total nitrogen (g·kg-1)	碳氮比C/N
常规中稻 Conventional middle rice (MR)	7.47±0.07a	136.17±8.19a	0.31±0.03a	13.61±3.41a	18.16±0.18a	1.86±0.01a	9.78±0.12a
稻虾共作 Rice-crayfish culture (CR)	7.41±0.15a	141.28±3.59a	0.40±0.01b	19.84±4.01a	18.59±0.14b	1.94±0.02b	9.58±0.10a
同列数据后不同字母表示差异显著(P < 0.05)。Values followed by different letters within the same column are significantly different (P < 0.05).

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表2稻虾共作模式和常规中稻模式3次重复土壤样品测序结果
Table2.Sequencing results of three repeated soil samples from integrated rice-crayfish farming system and conventional middle rice system

样品 Sample	序列数 Sequence number	碱基数 Base number	平均长度 Mean length (bp)	最短序列长度 Min. length (bp)	最长序列长度 Max. length (bp)
MR1	30 383	13 726 838	451.79	206	533
MR2	37 141	16 782 200	451.85	203	531
MR3	36 020	16 225 689	450.46	203	524
CR1	35 271	15 613 510	442.67	204	531
CR2	34 262	15 204 864	443.78	202	537
CR3	36 700	16 455 919	448.39	220	524
MR1、MR2和MR3分别表示常规中稻模式的3次重复, CR1、CR2和CR3分别表示稻虾共作模式的3次重复。MR1, MR2 and MR3 mean three replicates of the conventional middle rice; CR1, CR2 and CR3 mean three replicates of the rice-crayfish culture.

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表3稻虾共作模式(CR)和常规中稻模式(MR)Alpha多样性指数
Table3.Alpha diversity indices of integrated rice-crayfish farming system (CR) and conventional middle rice system (MR)

种植模式 Planting pattern	Sobs指数 Sobs index	Shannon指数 Shannon index	Simpson指数 Simpson index	Ace指数 Ace index	Chao指数 Chao index
MR	433.67±60.45a	3.99±0.33a	0.07±0.05a	492.07±50.19a	502.34±42.43a
CR	711.50±78.50b	4.59±0.24a	0.03±0.01a	750.25±72.39b	751.37±62.14b
同列数据后不同字母表示差异显著(P < 0.05)。Values followed by different letters within the same column are significantly different (P < 0.05).

下载: 导出CSV

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