徐峰3,
刘永胜4,
王学敏5,
李全1,
云兴福1,,
1.内蒙古农业大学农学院 呼和浩特 010018
2.赤峰学院生命科学学院 赤峰 024000
3.赤峰市敖汉旗农业技术服务中心 赤峰 024000
4.陕西省渭南市潼关县农业局园艺站 渭南 714300
5.内蒙古巴彦淖尔市农牧业科学研究院 巴彦淖尔 015000
基金项目: 国家自然科学基金项目31160100
内蒙古自治区应用技术研究与开发项目201602050
详细信息
作者简介:秦立金, 研究方向为设施蔬菜土传病害生物防治与土壤修复。E-mail:1597355169@qq.com
通讯作者:云兴福, 研究方向为高寒地区蔬菜栽培与生理。E-mail:yxf5807@163.com
中图分类号:S476计量
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被引次数:0
出版历程
收稿日期:2018-01-19
录用日期:2018-03-22
刊出日期:2018-08-01
Analysis of soil bacterial diversity under cucumber-celery intercropping and its influence on cucumber Fusarium wilt
QIN Lijin1, 2,,XU Feng3,
LIU Yongsheng4,
WANG Xuemin5,
LI Quan1,
YUN Xingfu1,,
1. College of Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China
2. College of Life Science, Chifeng University, Chifeng 024000, China
3. Agricultural Technical Service Center of Aohan Banner, Chifeng 024000, China
4. Horticultural Station of Tongguan County Agricultural Bureau in Weinan City, Shaanxi Province, Weinan 714300, China
5. Research Institute of Agriculture and Animal Husbandry in Bayannur City, Inner Mongolia, Bayannur 015000, China
Funds: the National Natural Science Foundation of China31160100
the Inner Mongolia Autonomous Region Application Technology Research and Development Project201602050
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Corresponding author:YUN Xingfu, E-mail:yxf5807@163.com
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摘要
摘要:本试验以黄瓜与西芹间作种植模式为处理，黄瓜单作和西芹单作种植模式为对照，利用Illumina公司Miseq平台对上述不同处理土壤进行16S rDNA细菌群落多样性高通量测序分析和田间接种黄瓜枯萎病菌，探讨黄瓜与西芹间作模式土壤细菌的多样性及其对田间黄瓜枯萎病发生的影响。16S rDNA测序结果表明，黄瓜与西芹间作土壤的细菌物种总数最多，群落多样性水平最高，与对照相比显著提高了土壤细菌observedspecies指数、Shannon指数和Chao1指数（P < 0.05）；Beta多样性聚类分析表明，黄瓜与西芹间作土壤的环境群落物种与黄瓜单作和西芹单作有一定差异性。在门分类水平上，共检测到45个菌门，其中变形菌门占明显优势，其次为酸杆菌门和放线菌门等；黄瓜与西芹间作土壤细菌种类所占比例最高，达98.63%。在属水平上，共检测到428类菌属，GP6、GP16、GP4、芽单胞菌属、节细菌属5属的丰度值较大；黄瓜与西芹间作土壤的节细菌属分布比例最高，红游动菌属、鞘氨醇单胞菌属和芽球菌属丰度值较大，为黄瓜与西芹间作土壤细菌明显优势菌属。田间接种黄瓜枯萎病菌试验结果表明，采用上述3种不同种植模式土壤种植黄瓜，在黄瓜苗期接种黄瓜枯萎病菌，黄瓜与西芹间作处理的黄瓜枯萎病的田间发病率较西芹单作和黄瓜单作分别降低57.03%~63.54%和66.95%~72.15%。因此，黄瓜与西芹间作增加了土壤细菌群落多样性，降低了黄瓜枯萎病的发病率，对后茬黄瓜土传病害防控具有一定科学指导意义。
关键词:黄瓜枯萎病/
黄瓜与西芹间作/
16S rDNA测序/
土壤细菌多样性/
田间发病率/
菌群丰度
Abstract:Cucumber wilt is a soil disease that is highly prevalent in the production and cultivation of cucumber. The disease is highly difficult to prevent and cure. A number of studies have shown that intercropping was one of the most effective methods of reducing the occurrence of plant soil diseases. Also celery has been proven to have high allelopathy. To explore the application of allelopathic effects of celery on cucumber fusarium wilt control, we conducted cucumber and celery intercropping experiment. In the experiment, three planting patterns were set, which were cucumber-celery intercropping, celery monocropping and cucumber monocropping. The cucumber-celery intercropping pattern was regarded as treatment group and monocropping patterns of celery and cucumber regarded as the control group. The Miseq platform of Illumina Company was used to analyze 16S rDNA bacterial community diversity through high-throughput sequencing to discuss the influence of cucumber-celery intercropping on cucumber soil bacterial diversity. Cucumber was planted in soil from different planting patterns and inoculated with Fusarium oxysporium f.sp.cucumerinum (Foc) in the pot experiment to investigate the control effect of cucumber-celery intercropping on cucumber fusarium wilt. The 16S rDNA sequencing results showed that total bacterial species amount and community diversity were highest in soil under cucumber-celery intercropping, which significantly enhanced observed bacteria species index, Shannon index and Chao1 index (P < 0.05). Beta diversity clustering analysis showed there existed a difference in environmental community species between soil from cucumber-celery intercropping and mono-cropped cucumber or celery. Moreover, 15 bacterial phyla were detected. Proteobacteria, which was followed by Acidobacteria and Actinobacteria, had a distinct advantage (35.7%-38.0%). Then the proportion of bacterial species derived from cucumber-celery intercropped soil was highest (98.63%). About 428 bacterial genera were detected with 5 dominant bacterial genera, which were GP6, GP16, GP4, Gemmatimonas and Arthrobacter. Arthrobacter, Rhodoplanes, Sphingomonas and Blastococcus were dominant bacteria genera in cucumber-celery inter-cropped soil. The 4 genera demonstrated that cucumber and celery intercropping enriched the diversity of bacterial communities compared with monocropped celery or cucumber. The results of fusarium wilt inoculation experiment of cucumber suggested that the control efficiency of cucumber-celery intercropping to cucumber fusarium wilt reached 57.03%-63.54% and 66.95%-72.15% relative to monocroped celery and cucumber, respectively. Therefore cucumber-celery intercropping increased the diversity of bacterial communities, and reduced incidence rate of cucumber fusarium wilt. This was of scientific interest for the prevention and control of soil borne diseases.
Key words:Cucumber fusarium wilt/
Cucumber-celery intercropping/
16S rDNA sequencing/
Soil bacterial diversity/
Field incidence/
Bacteria abundance
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图1不同处理土壤细菌OUT分布venn图
XD、HD、JZ分别代表西芹单作、黄瓜单作和黄瓜与西芹间作。
Figure1.Venn diagram illustrating OUT (Operational Taxonomy Unit) distribution of soil bacteria under different treatments
XD, HD, and JZ indicate celery monoculture, cucumber monoculture and cucumber-celery intercropping, respectively.
下载: 全尺寸图片幻灯片
图2不同处理样品土壤细菌群落的聚类树形图
XD、HD、JZ分别代表西芹单作、黄瓜单作和黄瓜与西芹间作。1、2和3为各处理的重复。
Figure2.Clustering tree diagram of bacteria communities of soil samples of different treatments
XD, HD, and JZ indicate celery monoculture, cucumber monoculture and cucumber-celery intercropping, respectively. 1, 2 and 3 show replicates of treatments.
下载: 全尺寸图片幻灯片
图3不同处理土壤细菌门水平的物种分类热图
XD、HD、JZ分别代表西芹单作、黄瓜单作和黄瓜与西芹间作。
Figure3.Heat map describing bacterial species classification at the phylum level under different treatments
XD, HD, and JZ indicate celery monoculture, cucumber monoculture and cucumber-celery intercropping, respectively.
下载: 全尺寸图片幻灯片
图4不同处理土壤细菌属水平上的群落组成
XD、HD、JZ分别代表西芹单作、黄瓜单作和黄瓜与西芹间作。
Figure4.Community composition of soil bacteria at the genus level under different treatments
XD, HD, and JZ indicate celery monoculture, cucumber monoculture and cucumber-celery intercropping, respectively.
下载: 全尺寸图片幻灯片
图5不同处理土壤细菌属水平上的物种分类热图
Figure5.Heat map describing bacterial species classification at the genus level under different treatments
下载: 全尺寸图片幻灯片表1不同处理土壤细菌Alpha多样性指数分析
Table1.Alpha diversity indexes of soil bacteria under different treatments
| 处理 Treatment | Observed species指数 Observed species index | Shannon指数 Shannon index | Chao1指数 Chao1 index | Simpson指数 Simpson index |
| 黄瓜与西芹间作 Cucumber-celery intercropping | 3 462.67±64.25a | 10.93±0.05a | 5 959.99±131.10a | 1 |
| 西芹单作Celery monoculture | 3 042.33±96.43c | 10.57±0.07b | 5 192.81±253.13c | 1 |
| 黄瓜单作Cucumber monoculture | 3 245.67±81.27b | 10.71±0.07b | 5 542.34±157.03b | 1 |
| 同列不同小写字母表示0.05水平差异显著。Data with different lowercase letters in the same column are significantly different at 0.05 level. | ||||
下载: 导出CSV表2Weighted unifrac法计算的不同处理样品土壤细菌群落间的距离
Table2.Intersample distances calculated using the method of weighted unifrac of bacteria communities of soil samples of different treatments
| 土样 Soil sample | HD1 | HD2 | HD3 | JZ1 | JZ2 | JZ3 | XD1 | XD2 | XD3 |
| XD1 | 0.17 | 0.14 | 0.15 | 0.13 | 0.13 | 0.16 | 0.00 | 0.12 | 0.15 |
| XD2 | 0.14 | 0.15 | 0.18 | 0.11 | 0.12 | 0.19 | 0.12 | 0.00 | 0.11 |
| XD3 | 0.15 | 0.16 | 0.16 | 0.13 | 0.14 | 0.19 | 0.15 | 0.11 | 0.00 |
| HD1 | 0.00 | 0.18 | 0.22 | 0.14 | 0.15 | 0.24 | 0.17 | 0.14 | 0.15 |
| HD2 | 0.18 | 0.00 | 0.16 | 0.14 | 0.15 | 0.18 | 0.14 | 0.15 | 0.16 |
| HD3 | 0.22 | 0.16 | 0.00 | 0.17 | 0.17 | 0.12 | 0.15 | 0.18 | 0.16 |
| JZ1 | 0.14 | 0.14 | 0.17 | 0.00 | 0.10 | 0.19 | 0.13 | 0.11 | 0.13 |
| JZ2 | 0.15 | 0.15 | 0.17 | 0.10 | 0.00 | 0.19 | 0.13 | 0.12 | 0.14 |
| JZ3 | 0.24 | 0.18 | 0.12 | 0.19 | 0.19 | 0.00 | 0.16 | 0.19 | 0.19 |
| XD、HD、JZ分别代表西芹单作、黄瓜单作和黄瓜与西芹间作。1、2和3为各处理的重复。XD, HD, and JZ indicate celery monoculture, cucumber monoculture and cucumber-celery intercropping, respectively. 1, 2 and 3 show replicates of treatments. | |||||||||
下载: 导出CSV表3不同处理土壤细菌在门水平上的群落组成
Table3.Community composition of soil bacteria at the phylum level under different treatments
| 菌门 Bacterial phylum | Read数 Reads | 比例 Ratio (%) | 菌门比例Phylum ratio (%) | ||
| XD | HD | JZ | |||
| 变形菌门Proteobacteria | 41 246 | 35.69 | 36.67±1.19a | 36.77±6.56a | 33.63±5.31b |
| 酸杆菌门Acidobacteria | 25 083 | 21.73 | 20.67±2.21a | 21.63±5.75a | 22.90±6.16a |
| 放线菌门Actinobacteria | 19 799 | 17.44 | 17.70±2.05b | 16.43±0.98b | 18.20±0.46a |
| 绿弯菌门Chloroflexi | 7 290 | 6.30 | 6.20±0.26b | 5.97±0.76b | 6.73±0.84a |
| 芽单胞菌门Gemmatimonadetes | 5 902 | 5.32 | 5.20±0.46b | 4.90±0.50b | 5.87±0.50a |
| 厚壁菌门Firmicutes | 5 354 | 4.60 | 4.97±0.51a | 4.57±0.67a | 4.27±0.95b |
| 拟杆菌门Bacteroidetes | 3 997 | 3.43 | 2.97±0.15b | 3.60±0.56a | 3.73±0.55a |
| 硝化螺旋菌门Nitrospirae | 2 088 | 1.77 | 1.73±0.40b | 1.67±0.46b | 1.90±0.26a |
| 浮霉菌门Planctomycetes | 1 047 | 0.90 | 0.80±0.10b | 1.03±0.21a | 0.87±0.25b |
| 疣微菌门Verrucomicrobia | 589 | 0.50 | 0.53±0.21a | 0.43±0.12b | 0.53±0.21a |
| Candidatus saccharibacteria | 543 | 0.48 | 0.43±0.06b | 0.43±0.06b | 0.57±0.15a |
| Latescibacteria | 232 | 0.18 | 0.13±0.06c | 0.23±0.06a | 0.17±0.12b |
| 迷踪菌门Elusimicrobia | 276 | 0.24 | 0.23±0.06b | 0.23±0.06b | 0.27±0.12a |
| OD1 | 140 | 0.19 | 0.13±0.06b | 0.13±0.06b | 0.17±0.06a |
| WS3 | 144 | 0.12 | 0.10±0.00a | 0.17±0.06a | 0.10±0.00a |
| 绿菌门Chlorobi | 118 | 0.11 | 0.10±0.00a | 0.10±0.10a | 0.13±0.06a |
| OP3 | 100 | 0.09 | 0.10±0.00a | 0.10±0.10a | 0.07±0.06a |
| TM7 | 70 | 0.07 | 0.07±0.06a | 0.07±0.06a | 0.07±0.06a |
| 无类别的Unclassified | 142 | 0.11 | 0.13±0.06a | 0.10±0.00a | 0.10±0.00a |
| 表中只列出门分类水平上菌群丰度百分比≥0.1%的菌门。同行不同小写字母表示0.05水平差异显著。XD、HD、JZ分别代表西芹单作、黄瓜单作和黄瓜与西芹间作。Only phyla with ≥0.1% abundance percentages were listed. Data with different lowercase letters in the same line are significantly different at 0.05 level. XD, HD, and JZ indicate celery monoculture, cucumber monoculture and cucumber-celery intercropping, respectively. | |||||
下载: 导出CSV表4不同处理土壤对黄瓜枯萎病发病及防治效果的影响
Table4.Effect of soils of different treatments on morbidity and control efficiency of cucumber fusarium wilt
| 处理 Treatment | 接种后7 d 7 d after inoculation | 接种后9 d 9 d after inoculation | 接种后11 d 11 d after inoculation | 接种后13 d 13 d after inoculation | 接种后15 d 15 d after inoculation | |||||||||
| 病情指数 Disease index | 防治效果 Control efficiency(%) | 病情指数 Disease index | 防治效果 Control efficiency(%) | 病情指数 Disease index | 防治效果 Control efficiency(%) | 病情指数 Disease index | 防治效果 Control efficiency(%) | 病情指数 Disease index | 防治效果 Control efficiency(%) | |||||
| 黄瓜与西芹间作 Cucumber-celery intercropping | 13.41c | — | 16.68c | — | 17.56c | — | 19.38c | — | 19.69c | — | ||||
| 西芹单作 Celery monoculture | 31.21b | 57.03b | 40.56b | 58.88b | 46.72b | 62.41b | 53.16b | 63.54b | 53.67b | 63.31b | ||||
| 黄瓜单作 Cucumber monoculture | 40.57a | 66.95a | 55.73a | 70.07a | 62.24a | 71.79a | 69.58a | 72.15a | 69.74a | 71.77a | ||||
| 同列不同小写字母表示0.05水平差异显著。Data with different lowercase letters in the same column are significantly different at 0.05 level. | ||||||||||||||
下载: 导出CSV参考文献
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