白羽祥^1,,
杨成翠¹,
史普酉¹,
贾孟¹,
杨焕文¹,
徐照丽²,
王戈^1,,
1.云南农业大学烟草学院 昆明 650201
2.云南省烟草农业科学研究院 昆明 650021
基金项目: 国家自然科学基金项目31860357
云南省应用基础研究计划项目2015FB145
云南省研究生学术新人奖A2008057
云南烟用有机肥养分资源利用研究2017YN07

详细信息

作者简介:白羽祥, 主要从事烟草生理生化研究。E-mail:cotsbyx@163.com

通讯作者:王戈, 主要从事烟草栽培生理生化研究。E-mail: wangge302@126.com

中图分类号:S572;S154.1

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出版历程

收稿日期:2018-07-27
录用日期:2018-10-15
刊出日期:2019-03-01

Correlation analysis of main environmental factors and phenolic acids in continuous tobacco cropping soils using Mantel Test

BAI Yuxiang^1,,
YANG Chengcui¹,
SHI Puyou¹,
JIA Meng¹,
YANG Huanwen¹,
XU Zhaoli²,
WANG Ge^1,,
1. College of Tobacco Science, Yunnan Agricultural University, Kunming 650201, China
2. Yunnan Academy of Tobacco Science, Kunming 650021, China
Funds: the National Natural Science Foundation of China31860357
the Applied Basic Research Project of Yunnan Province2015FB145
the Graduate Scholar Newcomer Award of Yunnan ProvinceA2008057
the Study on Utilization of Organic Fertilizer of Tobacco in Yunnan Province2017YN07

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Corresponding author:WANG Ge, E-mail: wangge302@126.com

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摘要
摘要:为明确酚酸类物质在连作植烟土壤中的变化特征，探讨土壤主要环境因子对酚酸类物质的影响，以不同连作年限（4 a，6 a，8 a，14 a和16 a）植烟土壤为对象，研究了不同连作年限植烟土壤酚酸类物质、理化性状、酶活性和细菌多样性的变化特征，并利用Mantel Test分析了酚酸类物质与土壤主要环境因子的相关性。结果表明，随连作年限增加，土壤酚酸类物质和速效钾含量升高，pH、有机质含量、细菌菌群丰度和多样性降低，水解性氮和有效磷含量呈先降低后升高趋势，酶活性呈先升高后降低趋势。Mantel Test分析表明，土壤酚酸类物质含量与理化性状、酶活性和细菌丰度显著相关，且与理化性状相关性最高；不同酚酸类物质含量与土壤主要环境因子相关性存在差异，其中，对羟基苯甲酸和阔马酸与植烟土壤理化性状、酶活性以及细菌丰度的相关性最高。因此，在本试验条件下，连作植烟土壤酚酸类物质具有明显积累特征，植烟土壤环境恶化；酚酸类物质积累受理化性状、酶活性和细菌多样性影响，且理化性状影响最大；不同酚酸类物质受主要土壤环境因子的影响存在差异，其中对羟基苯甲酸和阔马酸积累所受影响最大。
关键词:烤烟/
连作/
酚酸类物质/
土壤环境/
Mantel Test
Abstract:Flue-cured tobacco is a crop sensitive to continuous cropping. Continuous cropping over the long-term has seriously affected growth and production quality of flue-cured tobacco by affecting the soil environment. Phenolic acids can cause growth disorders under continuous cropping. Although the interaction between phenolic acids and other environmental factors after entering the soil environment has not much been investigated, it is clear that phenolic acids occur in soils under continuous tobacco cropping. The action process and the role in limiting continuous cropping are critical for successful tobacco cultivation. An experiment was conducted to determine the relationship between phenolic acids and soil environmental factors in the soil by analyzing the correlation between various environmental factors and phenolic acids in soil. The purpose was to identify the environmental factors affecting the concentration of phenolic acids in soils under continuous cropping systems. Soils of four tobacco fields with different durations of continuous cropping (4 a, 6 a, 8 a, 14 a and 16 a) were sampled. The changes in phenolic acids, physical and chemical properties, enzyme activities and bacterial diversities in soils under different continuous cropping years of tobacco were investigaed. The correlation between phenolic acids and soil environmental factors was analyzed using the Mantel Test. The results showed that long-term continuous cropping of tobacco resulted in an increase in soil phenolic acids content. Higher pH of soil under tobacco continuous cropping resulted in low soil organic matter content, but increased available potassium. The activities of catalase, urease acid phosphatase, and invertase first increased and then later decreased. When continuous cropping for less than 6 years, bacterial community structure changed little. However, bacterial community structure changed significantly after 8 years of continuous cropping. In general, continuous cropping changed bacterial community structure and reduced community diversity. Mantel Test analysis showed that phenolic acid content was significantly correlated with enzymes activities and physical and chemical properties of soil. Bacterial abundance had the highest correlation with soil physical and chemical properties. There were differences in correlation between different phenolic acids and soil environmental factors. P-hydroxybenzoic acid and phoronic acid had the highest correlation with physical and chemical properties of soil, enzyme activity and bacterial abundance in soils. Therefore, phenolic acids in soils under continuous tobacco cropping obviously accumulated over time with deteriorated soil environment. The accumulation of phenolic acids was affected by soil physical, chemical and biological properties of soil. P-hydroxybenzoic acid and tartaric acid were two kind phenolic acids most affected.
Key words:Flue-cured tobacco/
Continuous cropping/
Phenolic acid/
Soil environmental factor/
Mantel Test

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图1不同连作年限植烟土壤细菌群落的Shannon、Simpson、ACE和Chao1指数
不同小写字母代表不同连作年限在5%水平显著。
Figure1.Effect of continuous cropping years of tobacco on Shannon, Simpson, ACE and Chao1 index of soil bacterial community
Different lowercase letters represent significant differences among different continuous cropping years at 0.05 level.


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图2连作植烟土壤酚酸类物质与环境因子相关性
**表示在0.01水平显著相关。
Figure2.Correlation between phenolic acids content and soil environmental factors of continuously cropped tobacco
** indicates significant correlation at 0.01 level.


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图3基于Mantel Test的连作植烟土壤酚酸类物质与土壤环境因子相关性
**表示在0.01水平显著相关。P:间苯三酚; HA:阔马酸; PA:对羟基苯甲酸; VA:香草酸; V:香兰素; FA:阿魏酸; BA:苯甲酸; CA:肉桂酸; A:总量。
Figure3.Correlation between phenolic acids and soil environmental factors of continuously cropped tobacco based on Mantel Test
** indicates significant correlation at 0.01 level.P: phloroglucinol; HA: caramelic acid; PA: p-hydroxybenzoic acid; VA: vanillic acid; V: vanillin; FA: ferulic acid; BA: benzoic acid; CA: cinnamic acid; A: total amount.


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图4连作植烟土壤中与土壤环境因子相关性最高的前4种酚酸类物质的韦恩图
A:理化性状; B:酶活性; C:细菌门水平相对丰度。
Figure4.Venn diagram of the first four phenolic acids with the highest correlation with soil environmental factors of continuously cropped tobacco
A: soil physic-chemical properties; B: soil enzyme activity; C: relative abundances of bacterial phyla.


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表1不同连作年限烤烟土壤样品细菌基因测序数据预处理统计及质控
Table1.Data pre-processing statistics and quality control of bacterial gene sequencing of soil samples of tobacco continuously planted for different years

连作年限 Continuous cropping years (a)	原始下机序列 Raw reads	Clean reads中碱基数目 Base number of clean reads	Clean reads平均长度 Average length of clean reads (nt)	Clean reads中质量值大于20的碱基占比 Q20	Clean reads中 GC碱基含量 GC proportion of clean reads (%)	Clean reads的序列占比 Effective of clean reads (%)
4	50 798	12 276 030	253	99	55	96
6	56 998	13 741 732	253	99	55	95
8	65 290	15 845 683	253	99	56	96
14	52 905	12 438 514	260	99	57	90
16	59 980	14 595 832	253	99	53	96

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表2不同连作年限植烟土壤酚酸类物质种类和含量
Table2.Species and contents of phenolic acids in soil of tobacco continuously cropped for different years

mg·kg-1
酚酸类物质种类 Species of phenolic acid	连作年限 Continuous cropping years (a)
	4	6	8	14	16
间苯三酚 Phloroglucinol	1.213±0.021e	1.320±0.098d	1.447±0.061c	1.583±0.156b	1.987±0.055a
阔马酸 Caramelic acid	0.050±0.010c	0.049±0.007c	0.058±0.010b	0.063±0.013b	0.134±0.015a
对羟基苯甲酸 P-hydroxybenzoic acid	0.127±0.005d	0.187±0.020c	0.122±0.010d	0.279±0.021b	0.467±0.020a
香草酸 Vanillic acid	0.073±0.011d	0.123±0.006c	0.089±0.010d	0.145±0.024b	0.278±0.0123a
香兰素 Vanillin	0.090±0.017d	0.148±0.030b	0.128±0.019c	0.166±0.015b	0.291±0.010a
阿魏酸 Ferulic acid	0.032±0.011c	0.044±0.023c	0.122±0.025b	0.132±0.007b	0.318±0.020a
苯甲酸 Benzoic acid	0.046±0.008d	0.079±0.005b	0.111±0.004a	0.065±0.002c	0.071±0.003bc
肉桂酸 Cinnamic acid	0.024±0.005b	0.025±0.002b	0.039±0.001a	0.010±0.002c	0.008±0.001d
总量 Total amount	1.684±0.054d	1.954±0.056c	2.105±0.060c	2.412±0.067b	3.535±0.150a
不同小写字母代表不同连作年限在5%水平显著。Different lowercase letters represent significant differences among different continuous cropping years at 0.05 level.

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表3不同连作年限植烟土壤理化性状的影响
Table3.Effect of continuous cropping years of tobacco on soil physico-chemical properties

性质 Property	连作年限 Continuous cropping years (a)
	4	6	8	14	16
pH	7.63±0.15a	7.40±0.13b	7.30±0.18b	6.61±0.12c	6.40±0.14d
有机质含量 Organic matter content (g·kg-1)	55.02±2.22a	51.92±2.71b	36.61±1.33c	34.88±3.17c	34.95±2.56c
水解性氮含量 Hydrolyzed nitrogen content (mg·kg-1)	201.5±10.3b	173.3±15.2b	120.1±15.0c	118.2±20.2c	330.5±10.8a
有效磷含量 Available phosphorus content (mg·kg-1)	51.04±3.59a	35.46±3.11b	19.86±0.80c	18.16±1.04c	22.26±2.05c
速效钾含量 Available potassium content (mg·kg-1)	466.7±25.1b	468.4±40.1b	463.4±20.8b	1 273.7±105a	1 260.4±110a
不同小写字母代表不同连作年限在5%水平显著。Different lowercase letters represent significant differences among different continuous cropping years at 0.05 level.

下载: 导出CSV
表4不同连作年限植烟土壤酶活性的变化
Table4.Effect of continuous cropping years of tobacco on soil enzymes activities

mg·g-1·(24h)-1
酶 Enzyme	连作年限 Continuously cropping years (a)
	4	6	8	14	16
过氧化氢酶 Catalase	3.81±0.13c	4.35±0.09b	4.92±0.10a	2.63±0.14d	2.03±0.16d
脲酶 Urease	0.61±0.03b	0.68±0.09a	0.63±0.04b	0.58±0.06c	0.45±0.02d
酸性磷酸酶 Acid phosphatase	1.50±0.08d	2.37±0.08b	2.64±0.10a	1.93±0.07c	1.32±0.17d
蔗糖酶 Invertase	13.80±0.27b	16.70±0.33a	16.30±0.84a	13.90±0.31b	12.30±0.29c
不同小写字母代表不同连作年限在5%水平显著。Different lowercase letters represent significant differences among different continuous cropping years at 0.05 level.

下载: 导出CSV
表5不同连作年限植烟土壤中门水平细菌的相对丰度
Table5.Relative abundances of bacterial phyla in soil of tobacco continuously cropped for different years

%
门 Phyla	连作年限 Continuously cropping years (a)
	4	6	8	14	16
变形菌门 Proteobacteria	59.6±2.5a	57.7±2.6a	48.0±1.8ab	37.3±1.7b	47.0±8.0ab
拟杆菌门 Bacteroidetes	11.0±0.6b	12.8±2.4b	6.5±0.7c	4.8±1.3c	35.2±3.1a
芽单胞菌门 Gemmatimonadetes	3.2±0.7b	2.5±0.2b	3.1±0.5b	13.3±1.0a	0.8±0.3b
放线菌门 Actinobacteria	11.3±2.4b	12.4±2.0b	20.7±1.6a	12.7±2.1b	6.5±2.4c
酸杆菌门 Acidobacteria	5.5±0.8bc	6.9±2.1b	9.5±0.3ab	13.2±4.4a	2.1±0.8c
硬壁菌门 Firmicutes	2.5±0.4b	1.3±0.2b	0.8±0.2b	2.1±0.9b	6.1±1.4a
绿弯菌门 Chloroflexi	1.4±0.1c	1.1±0.2c	3.5±0.3b	5.3±1.6a	0.4±0.1c
疣微菌门 Verrucomicrobia	1.4±0.3b	1.6±0.5b	3.3±0.8a	3.6±0.4a	0.6±0.2c
浮霉状菌门 Planctomycetes	0.7±0.2ab	1.0±0.4ab	1.5±0.6ab	1.7±1.2a	0.3±0.1b
硝化螺旋菌门 Nitrospirae	1.3±0.1a	0.9±0.2b	0.7±0.1b	1.3±0.3a	0.4±0.1a
其他 Others	2.2±0.2b	1.9±0.2b	2.5±0.3b	4.8±0.5a	0.8±0.3c
不同小写字母代表不同连作年限在5%水平显著。Different lowercase letters represent significant differences among different continuous cropping years at 0.05 level.

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