Effects of sludge addition on bacterial diversity in rhizosphere and non-rhizosphere calcareous soil
CHANG Huiqing1,,, ZHENG Caijie1, LI Zhaojun2, JIAO Changfeng1, ZHU Xiaohui1, HE Wanling3, WANG Qizhen1, WU Jie1 1.School of Agriculture, Henan University of Science and Technology, Luoyang 471000, China 2.Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China 3.School of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471000, China
Abstract:In order to study the effects of sludge application on bacterial diversity of calcareous soil, different amounts of compost sludge were applied in a 2 consecutive year-summer maize field system. The high-throughput sequencing of 16S rDNA-V3-V4 region was carried out for collected rhizosphere and non-rhizosphere soil. Effects of sludge application on bacterial richness, diversity index, community formation and functional genes were analyzed. The correlation between bacterial community and environmental factors was also determined. The results showed that the amounts of OTU in these two kinds of soil increased at sludge addition amounts of 3.5~37.5 t·hm?2 in non-rhizosphere soil and 3.5~7.5 t·hm?2 in rhizosphere soil, respectively. However, at sludge addition amounts above 75 t·hm?2, Shannon index of bacterial diversity in rhizosphere and non-rhizosphere soil decreased significantly, Simpson index increased significantly, Ace and Chao indices for bacterial abundance also decreased significantly. Principal component and phylum-level species abundance analysis showed that different sludge addition amount could cause the differences in rhizosphere and non-rhizosphere bacterial community structure, and the abundance of actinobacteria in non-rhizosphere soil was significantly reduced at the sludge addition amount of 75 t·hm?2. The abundance of Bacteroidetes in non-rhizosphere soil increased significantly at the sludge addition amounts of 3.75~37.5 t·hm?2. Compared with the control, at the sludge addition amount of 75 t·hm?2, the abundance of Bacteroidetes in rhizosphere soil increased by 1.45 times, but the abundances of Acidobacteria and Actinobacteria decreased by 49.74% and 80.57%, respectively. Redundancy analysis (RDA) showed that soil TN, Cd, Cu and TP were the most important factors affecting soil bacterial community, among which TN, Cd, Cu were significantly correlated with Shannon and Simpson diversity indexes (P<0.05). Therefore, the continuous and excessive sludge application can adversely affect bacterial diversity in rhizosphere and non-rhizosphere calcareous soil, Moreover, the changes of microbial diversity indices mentioned above can be used as a basis for the rational sludge application. Key words:sludge/ calcareous soil/ rhizosphere/ non-rhizosphere/ bacterial diversity.
图1污泥施用对非根际和根际土壤OTU丰度影响 Figure1.Effects of sludge application on OTU abundance in non-rhizosphere and rhizosphere soil
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1.School of Agriculture, Henan University of Science and Technology, Luoyang 471000, China 2.Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China 3.School of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471000, China Received Date: 2019-01-20 Accepted Date: 2019-04-15 Available Online: 2019-09-17 Keywords:sludge/ calcareous soil/ rhizosphere/ non-rhizosphere/ bacterial diversity Abstract:In order to study the effects of sludge application on bacterial diversity of calcareous soil, different amounts of compost sludge were applied in a 2 consecutive year-summer maize field system. The high-throughput sequencing of 16S rDNA-V3-V4 region was carried out for collected rhizosphere and non-rhizosphere soil. Effects of sludge application on bacterial richness, diversity index, community formation and functional genes were analyzed. The correlation between bacterial community and environmental factors was also determined. The results showed that the amounts of OTU in these two kinds of soil increased at sludge addition amounts of 3.5~37.5 t·hm?2 in non-rhizosphere soil and 3.5~7.5 t·hm?2 in rhizosphere soil, respectively. However, at sludge addition amounts above 75 t·hm?2, Shannon index of bacterial diversity in rhizosphere and non-rhizosphere soil decreased significantly, Simpson index increased significantly, Ace and Chao indices for bacterial abundance also decreased significantly. Principal component and phylum-level species abundance analysis showed that different sludge addition amount could cause the differences in rhizosphere and non-rhizosphere bacterial community structure, and the abundance of actinobacteria in non-rhizosphere soil was significantly reduced at the sludge addition amount of 75 t·hm?2. The abundance of Bacteroidetes in non-rhizosphere soil increased significantly at the sludge addition amounts of 3.75~37.5 t·hm?2. Compared with the control, at the sludge addition amount of 75 t·hm?2, the abundance of Bacteroidetes in rhizosphere soil increased by 1.45 times, but the abundances of Acidobacteria and Actinobacteria decreased by 49.74% and 80.57%, respectively. Redundancy analysis (RDA) showed that soil TN, Cd, Cu and TP were the most important factors affecting soil bacterial community, among which TN, Cd, Cu were significantly correlated with Shannon and Simpson diversity indexes (P<0.05). Therefore, the continuous and excessive sludge application can adversely affect bacterial diversity in rhizosphere and non-rhizosphere calcareous soil, Moreover, the changes of microbial diversity indices mentioned above can be used as a basis for the rational sludge application.
供试污泥和采集的土壤样品风干过筛后分析以下指标：全氮，全磷，全钾、有机质、pH、电导率。上述指标测定方法参考文献中的方法[25]。重金属测定：取0.500 g通过100目筛样品，置于聚氟坩埚中，加入10 mL HF，8 mL HNO3和3 mL HClO4后静置12 h，以除去有机质和硅酸，然后在加热板上加热，温度逐渐升高至200 ℃，稳定温度加热至白烟产生且坩埚中残存少量浅白色黏稠状物质为止。完全冷却后，加2%的HNO3 2~3 mL溶解残存物质，用25 mL容量瓶定容待用，空白和标准样品同时消化，样品中的Zn、Cu、Pb、Cr、Cd利用等离子发射光谱法(美国，PE optima5300DV)测定。As采用双道原子荧光光度计(北京，吉田AFS-9120)测定。每个样品重复3次，使用SPSS 19.0软件进行单因素方差分析和相关性分析。
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