Soil Microbiome at Phyllostachys edulis Forest Affected by Application of Bioagent
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摘要:
目的 探究复合微生物菌剂[产气肠杆菌（Enterobacter aerogenes）CT-B09-2、解淀粉芽孢杆菌（Bacillus amyloliquefaciens）JL-B06和乙酸钙不动杆菌（Acinetobacter calcoaceticus）WYS-A01-1]对毛竹土壤细菌群落结构和多样性的影响。 方法 以毛竹（Phyllostachys edulis）实生幼苗为研究对象，采用灌根的方式施用复合微生物菌剂，30 d后采集毛竹幼苗根际与非根际土壤样本，测定土壤理化性质，提取土壤总DNA并进行16S rRNA高通量测序，分析复合微生物菌剂对土壤细菌群落结构和多样性的影响。 结果 复合微生物菌剂可以有效提升土壤中速效磷的含量，调控土壤pH，提高根际土壤中物质代谢和碳化合物分解相关的功能活性。毛竹根际与非根际土壤共检测出26门、65纲、158目、253科、448属、674种微生物，主要优势菌门包括变形菌门 Proteobacteria、拟杆菌门 Firmicutes、放线菌门 Actinobacteriota、酸杆菌门 Acidobacteriota、绿弯菌门 Chloroflexi等。施用复合微生物菌剂后，毛竹根际土壤微生物群落物种数目显著上升，非根际土壤样本无显著变化。 结论 复合微生物菌剂可以调节土壤矿质元素，改善土壤pH，调控细菌微生物群落组成。 Abstract:Objective Effect of application of a microbial agent on the microbiome in soil at a Phyllostachys edulis forest was analyzed. Method In a P. edulis forest, a microbial solution of Enterobacter aerogenes CT-B09-2, Bacillus amyloliquefaciens JL-B06, and Acinetobacter calcoaceticus WYS-A01-1 was applied by root irrigation onto the soil around the seedlings. Rhizosphere and non-rhizosphere soil samples at the spots were collected 30 d after the application for physical and chemical analyses. Total DNA in soil was extracted, and 16S rRNA high-throughput sequencing performed to dissect the microbial community structure and diversity. Results The applied bioagent effectively increased the available phosphorus, regulated the pH, and improved the functional activities related to material metabolism and decomposition of carbon compounds in the rhizosphere soil. There were 26 phyla, 65 classes, 158 orders, 253 families, 448 genera, and 674 species detected in the soil samples with Proteobacteria, Firmicutes, Actinobacteriota, Acidobacteriota, and Chloroflexi being the dominant phyla. The application significantly increased the number of species in the microbial community only in the rhizosphere soil. Conclusion Application of the mixed microbial cultures significantly and positively affected the mineral contents, pH, and microbial community in the rhizosphere soil at the bamboo forest. -
Key words:
- bioagent /
- Phyllostachys edulis /
- soil microbiome /
- diversity
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图 1 样本稀释曲线和Venn图
A：稀释曲线；B：（a）复合微生物菌剂施用的根际土壤ASV对比；（b）复合微生物菌剂施用的非根际土壤ASV对比；（c）样本总体ASV对比。
Figure 1. Sample dilution curve and Venn diagram
A: dilution curve; B: (a) ASVs of rhizosphere soils treated with bioagent; (b) ASVs of non-rhizosphere soils treated with bioagent; (c) ASVs of total sample population.
图 3 门水平下的物种相对丰度
A：相对丰度；B：样本间显著差异箱图。图4同。
Figure 3. Relative species abundance at phylum level
A: histogram of relative abundance; B: box chart of significant differences between samples. Same for Fig.4.
图 5 样本环境因子与门水平物种RDA分析
Figure 5. RDA analysis on environmental factors and species at phylum level
表 1 样本编号对照表
Table 1. Sample codes
样本类型
Sample type分组
Group对照组毛竹根际土壤 E0 对照组毛竹非根际土壤 F0 复合微生物菌剂处理组毛竹根际土壤 E1 复合微生物菌剂处理组毛竹非根际土壤 F1 
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表 2 土壤理化性质测定结果
Table 2. Physiochemical properties of soil samples
样本
Sample有机质
OM/ (g·kg−1)速效磷
AP/（μmol·g−1）速效钾
AK/（mg·kg−1）全钾
TK/（g·kg−1）全氮
TN/（g·kg−1）全磷
TP/（mg·kg−1）速效氮
AN/（mg·kg−1）pH E0 20.81±0.01a 4.42±0.02c 329.12±0.02b 14.79±0.02b 0.97±0.02a 507.04±0.02b 86.34±0.01a 5.68±0.01d E1 7.67±0.01c 4.82±0.03a 196.71±0.01d 13.75±0.02c 0.75±0.03b 480.41±0.03c 43.03±0.02c 5.88±0.01c F0 17.84±0.01b 4.50±0.01b 341.53±0.03a 15.84±0.04a 0.99±0.01a 531.07±0.04a 68.96±0.04b 6.14±0.02b F1 7.01±0.01d 4.51±0.01b 261.03±0.04c 13.73±0.03c 0.77±0.02b 488.1±0.02c 47.05±0.03c 6.54±0.02a 图中数据为平均值±标准差；同列数据后不同小写字母表示不同处理间差异显著（P ＜ 0.05）。表3同。
Data are mean±standard deviation; those with different lowercase letters indicate significant differences between treatments at P＜0.05. Same for Table 3.
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表 3 Alpha 多样性指数
Table 3. Alpha diversity index
样本
Sample丰度指数
ACE查尔指数
Chao1香农指数
Shannon辛普森指数
SimpsonE0 479.84±32.70b 479.76±32.55b 5.74±0.11a 0.0048±0.001b E1 539.52±80.66a 539.20±80.37a 5.76±0.06a 0.0055±0.001b F0 493.95±96.69b 493.66±96.37b 5.67±0.19a 0.0054±0.001b F1 489.88±127.07b 489.25±126.73b 5.56±0.29a 0.0079±0.004a
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表 4 细菌功能表达丰度
Table 4. Significant differences in microbial functions
功能 Function E0 E1 F0 F1 光养 Phototrophy 1.54±0.37c 1.61±0.09b 2.62±0.26a 2.32±0.26a 光合自养 Photoautotrophy 1.47±0.38d 1.61±0.09c 2.60±0.29a 2.26±0.32b 蓝藻 Cyanobacteria 1.23±0.36d 1.55±0.04c 2.56±0.35a 2.26±0.32b 含氧光能自养 Oxygenic photoautotrophy 1.23±0.36d 1.55±0.04c 2.56±0.35a 2.26±0.32b 固氮作用 Nitrogen fixation 3.98±1.90a 1.94±0.20b 0.75±0.31c 0.67±0.36c 硝酸盐呼吸 Nitrate respiration 0.98±0.58a 0.26±0.12c 0.84±0.32b 0.20±0.09c 氮呼吸 Nitrogen respiration 0.98±0.58a 0.26±0.12b 0.84±0.32a 0.20±0.09b 纤维素分解 Cellulolysis 1.04±0.50a 0.50±0.32b 0.16±0.02c 0.15±0.16c 甲基营养 Methylotrophy 0.19±0.01a 0.05±0.08c 0.19±0.02a 0.08±0.05b 甲醇氧化 Methanol oxidation 0.19±0.01a 0.05±0.08c 0.19±0.02a 0.08±0.05b 铁呼吸 Iron respiration 0d 0.02±0.03c 0.14±0.04a 0.07±0.07b 芳香烃碳氢降解 Aromatic hydrocarbon degradation 0c 0.02±0.02b 0c 0.09±0.07a 脂族非甲烷碳氢化合物降解
Aliphatic non methane hydrocarbon degradation0c 0.02±0.02b 0c 0.09±0.07a 碳氢化合物降解 Hydrocarbon degradation 0c 0.02±0.02b 0c 0.09±0.07a
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