基于高通量测序的穿心莲连作根际土壤细菌群落多样性分析
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广东省科技计划(2016A020226050);广东省重点领域研发计划(2020B020221002)


Analysis of bacterial community diversity in rhizosphere soil of continuous cropping Andrographis paniculata based on high-throughput sequencing
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    摘要:

    目的 研究穿心莲Andrographis paniculata连作对土壤细菌多样性及种群结构的影响。方法 通过16S rRNA 基因高通量测序对连作组(连作5年穿心莲的根际土壤)和对照组(连作5年穿心莲周边相同土质的未耕作自然土壤)样品进行测序分析,比较2组土壤细菌的丰富度、多样性和群落结构差异。结果 α 多样性指数分析结果显示,连作组土壤中细菌丰富度指数Chao1指数、Observed species指数显著低于对照组(P<0.05),连作组土壤中的多样性指数Shannon指数和Simpson指数均低于对照组,但差异不显著。主坐标分析及分子方差分析进行组间群落结构差异显著性分析结果显示,穿心莲连作显著改变了土壤细菌群落结构(P<0.05)。从2个处理组土壤样品中共检测到2769个可操作分类单元,分属于47门885属,穿心莲连作明显改变了土壤细菌在门和属水平上的结构分布。在门分类水平上,酸杆菌门Acidobacteria、变形菌门Proteobacteria、拟杆菌门Bacteroidetes和Patescibacteria等11个细菌门是2个处理组的主要细菌类群,连作组土壤中变形菌门和酸杆菌门的相对丰度较对照组分别增加了21.97%和46.33% (P<0.05),而Patescibacteria较对照组减少了68.99% (P<0.05);在属分类水平上,有10个优势菌属的相对丰度变化显著(P<0.05),其中连作组土壤中有益生防细菌黄杆菌属FlavobacteriumHaliangium的相对丰度较对照组分别减少了78.40%和54.55%,而植物病原细菌伯克氏菌属Burkholderia-Caballeronia-Paraburkholderia的相对丰度较对照组增加了804.17%。结论 穿心莲5年连作降低了土壤细菌丰富度和多样性水平,使土壤微生态系统稳定性降低,同时有益菌相对丰度显著降低,而病原菌相对丰度显著增加造成细菌群落结构改变。上述变化打破了原有的土壤微生态平衡,可能是穿心莲连作障碍的重要原因之一。

    Abstract:

    Objective To investigate the effects of continuous cropping of Andrographis paniculata on soil bacterial diversity and community structure.Method Sequencing analysis of samples from continuous cropping group (rhizosphere soil of A. paniculata continuous cropping for five years) and control group (uncultivated natural soil with the same texture as the soil around A. paniculata continuous cropping for five years) were performed using 16S rRNA gene high-throughput sequencing technology. The differences in richness, diversity and community structure of soil bacteria from two groups were analyzed.Result The analysis results of α diversity index showed that the bacterial richness indices (Chao1 index and Observed species index) in the soil of continuous cropping group were significantly lower than those of the control group (P<0.05), and the diversity indices (Shannon index and Simpson index) in the soil of continuous cropping group were lower than those of the control group, but the differences were not significant. Principal coordinates analysis and molecular variance analysis of community structure between groups showed that continuous cropping of A. paniculata significantly changed soil bacterial community structure (P<0.05). A total of 2769 operational taxonomic units were detected from the soil samples of two groups, belonging to 47 phyla and 885 genera. Continuous cropping of A. paniculata obviously changed the structure distribution of soil bacteria at the phylum and genus level. At the phylum classification level, 11 bacterial phyla including Acidobacteria, Proteobacteria, Bacteroidetes, Patescibacteria and etc. were the main bacterial communities in two groups. Compared with the control group, the relative abundance of Proteobacteria and Acidobacteria in the continuous cropping group increased by 21.97% and 46.33% respectively (P<0.05), while the relative abundance of Patescibacteria reduced by 68.99% (P<0.05). At the genus classification level, the relative abundance of 10 dominant bacterial genera changed significantly (P<0.05). Among them, compared with the control group, the relative abundance of the beneficial bacteria Flavobacterium and Haliangiumin in the soil of continuous cropping group reduced by 78.40% and 54.55% respectively, while the relative abundance of the plant pathogenic bacterium Burkholderia-Caballeronia-Paraburkholderia in the soil of continuous cropping group increased by 804.17%.Conclusion After five years of A. paniculata continuous cultivation, the level of soil bacterial richness and diversity decreased. At the same time, the relative abundance of beneficial bacteria significantly reduced, while the relative abundance of pathogenic bacteria significantly increased, causing a change in the bacterial community structure. The above-mentioned changes broke the original soil microecological balance, which may be one of the important reasons for A. paniculata continuous cropping obstacles.

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周界,李明,徐友阳,洪彪,段书蕾,张慧晔,邹江林.基于高通量测序的穿心莲连作根际土壤细菌群落多样性分析[J].华南农业大学学报,2021,42(3):55-63

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  • 收稿日期:2020-06-25
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  • 在线发布日期: 2021-04-28