Endophytic and Rhizosphere Microbes in Pinellia ternata and Habitat Soils in Guizhou Affected by Environmental Conditions
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摘要:
目的 探究贵州不同生境半夏内生和根际土壤微生物群落多样性、物种组成、物种网络关系和功能菌群预测，并分析其与环境因子的相关性，为半夏菌群资源开发利用提供理论依据。 方法 对贵州野生半夏、规模化种植半夏和套种半夏的内生菌群、根际土壤菌群进行高通量测序，检测土壤理化因子和酶活性，并进行生物信息学分析。 结果 人工种植和野生半夏土壤环境因子差异性明显，且栽培方式对土壤理化性质与酶活有显著影响，人工种植样地的有机质含量高且土壤酸性强，野生样地酶活性偏高；半夏内生细菌优势菌属包括根瘤菌属（Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium）、假单胞菌属（Pseudomonas）、芽孢杆菌属（Bacillus）、慢生根瘤菌属（Bradyrhizobium）等，半夏内生真菌优势菌属包括弯孢菌属（Campylospora）、新赤壳属（Neocosmospora）、丝核菌属（Rhizoctonia）、镰刀菌属（Fusarium）、Dactylonectria、炭角菌目（Xylariales）未分类属等，半夏根际优势菌属种类更丰富，各优势菌属在不同样本以不同丰度聚集且具有规律性，受生长环境和种植方式影响；细菌优势菌群中有10属、真菌优势菌群中有13属，与土壤理化性质或酶活性呈显著相关性，内生菌群对环境因子更为敏感，半夏内生菌群既互利又拮抗，根际土壤菌群网络关系相对稳定；半夏内生菌群优势菌属中除了未明确分类、未知菌属外，有益菌属丰度占比高。 结论 可通过人为调节土壤pH、施入菌肥等方式改良土壤微生态环境；半夏内生菌群包括根瘤菌、假单胞菌、芽孢杆菌、农研丝杆菌、木霉菌和绿僵菌等可开发利用，助力半夏栽培，促进半夏产业发展。 Abstract:Objective Endophytic microbes of Pinellia ternata and rhizosphere microbial communities in the habitat soils in Guizhou as affected by the environmental conditions were analyzed. Methods High throughput sequencing to identify the endophytic and rhizosphere microbiota in P. ternata and soils of the wild, large-scale cultivated, and intercropped P. ternata plants grew in Guizhou was conducted. Microbial diversity, species, relationships, and enzyme activities as well as functional microflora were analyzed. Bioinformatics was employed to decipher the ecological relationship between the plant and its habitat. Results The environmental, physiochemical, and enzymatic conditions on the land where P. ternata plants were grown in the wild or under cultivation varied significantly. For instance, the organic matter content and acidity were high in the cultivated plots, but the enzyme activity was high in the soil of virgin forest. In the plants, Rhizobium, Pseudomonas, Bacillus, and Bradyrhizobium were the dominant endophytic bacteria genera, whereas Campylospora, Neocosmospora,Rhizoctonia, Fusarium, Dactylonectria, unclassified genera of Xylariales order, etc. were found to be the dominant fungi genera. In the rhizosphere soil, abundant dominant microbial species regularly aggregated and were affected by the environmental factors and planting method. For example, 10 dominant bacteria genera and 13 fungi genera significantly correlated with the physicochemical properties and/or enzyme activity of the soil they inhabited. In general, the endophytic microbes in the plants were more sensitive to the environmental factors than those in the rhizosphere soil. And they could be synergistic as well as antagonistic to one another, but the rhizosphere community tended to be stable. Aside from the unclassified and unknown genera, the dominant endophytic bacteria were mostly considered beneficial for the plant. Conclusion The microbial community in the soil could be improved by adjusting pH and applying microbial fertilizers. The endophytic microbes of P. ternate including Rhizobia,Pseudomonas, Bacillus, Agrobacterium, Trichoderma, and Metarhizium anisopliae could be used to enhance plant growth ushering in the development of a pharmaceutical industry based on the medicinal material. -
Key words:
- Pinellia ternata /
- endophyte /
- soil microbes /
- environmental factor
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图 1 不同生境半夏细菌OTU水平韦恩图
Figure 1. Venn plot of bacterial OTUs in P. ternata habitat soils
图 3 不同生境半夏细菌群落Shannon指数差异
Figure 3. Bacterial Shannon indexes of P. ternata habitat soils
图 4 不同生境半夏真菌菌群Shannon指数差异
* 表示差异显著（P ≤ 0.05） 。
Figure 4. Fungal Shannon indexes of P. ternata habitat soils
* indicate significant difference ( P ≤ 0.05).
图 5 不同生境半夏细菌OTU水平NMDS分析
Figure 5. NMDS analysis on bacterial OTUs of P. ternata habitat soils
图 6 不同生境半夏真菌OTU水平NMDS分析
Figure 6. NMDS analysis on fungal OTUs of P. ternata habitat soils
图 7 不同生境半夏细菌门水平物种组成
Figure 7. Bacteria species at phylum level in P. ternata habitat soils
图 8 不同生境半夏真菌门水平物种组成
Figure 8. Fungi species at phylum level in P. ternata habitat soils
图 9 不同生境半夏细菌属水平物种组成
Figure 9. Bacteria species at genus level in P. ternata habitat soils
图 10 不同生境半夏真菌属水平物种组成
Figure 10. Fungi species at genus level in P. ternata habitat soils
图 11 细菌属水平物种间相关性分析
圆点的大小表示物种之间相关性强度，下同。
Figure 11. Correlations among bacteria species at genus level
Size of dot indicates correlation between specie. Same for below.
图 16 不同生境半夏细菌属水平物种与环境因子相关性
Figure 16. Correlation between bacteria species and environmental factors in P. ternata habitat soils
图 17 不同生境半夏真菌属水平物种与环境因子相关性
Figure 17. Correlation between fungi species at genus level and environmental factors in P. ternata habitat soils
表 1 半夏采样信息
Table 1. Sampling of P. ternata
编号
Code采集地点
Locality采集部位和对应编号
Collection location and coding生长环境
Growth environment经度
Longitude/ （°）纬度
Latitude/（°）海拔
Altitude/mDH 毕节市大河乡 块茎和根须（DHK）、根际土（DHT） 林下野生 27.319420 104.920517 1638.5 HZ 毕节市赫章县半夏种植基地 块茎和根须（HZK）、根际土（HZT） 大田人工起垄栽培 27.074467 104.420799 2216.4 DF 毕节市百里杜鹃
管委会鹏程管理区块茎和根须（DFK）、根际土（DFT） 滇重楼套种，人工起垄栽培，
种植基地周边为针阔混交林27.232323 105.785965 1633.7 
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表 2 不同生境半夏土壤理化性质及酶活性
Table 2. Physiochemical properties and enzyme activities of P. ternata habitat soils
指标
Index野生DH 规模化种植HZ 套种DF 总氮TN/（g·kg−1） 1.45±0.01b 1.63±0.03a 1.42±0.02b 总磷TP/（g·hg−1） 0.08±0.01c 0.10±0.00b 0.21±0.00a 总钾TK/（g·hg−1） 1.45±0.03a 1.11±0.00b 1.01±0.02c 有效氮AN/（mg·kg−1） 207.73±4.68c 301.13±2.85a 234.86±0.43b 有效钾AK/（mg·kg−1） 462.84±1.08a 368.87±1.69c 398.78±1.35b 有效磷AP/（mg·kg−1） 44.23±0.44c 47.71±0.62b 76.82±0.42a 有机质OM/（g·kg−1） 44.64±0.78c 73.14±0.60a 55.33±0.23b pH 6.69±0.03b 5.45±0.02c 6.84±0.02a 土壤脲酶 URE/（U·g−1） 0.58±0.03b 0.83±0.08a 0.64±0.07b 蔗糖酶 SC/（U·g−1） 20.99±1.64a 10.47±0.43b 11.05±0.89b 酸性磷酸酶 ACP/（U·g−1） 2.35±0.06a 0.82±0.07c 1.52±0.10b 多酚氧化酶 PPO/（U·g−1） 3.80±0.13a 0.88±0.07c 1.79±0.03b 同行中数值后面的不同小写字母表示处理间差异显著（P＜0.05）。
Data with different lowercase letters on same row indicate significant differences (P＜0.05).
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