JOURNAL OF NATURAL RESOURCES >
Effects of corn-soybean rotation system on soil nitrogen-fixing bacteria community structure and quality in black soil
Received date: 2021-11-01
Revised date: 2022-04-07
Online published: 2022-12-28
The effects of soybean continuous cropping (CS), maize continuous cropping (CM), corn-soybean rotation (MS), corn-corn-soybean (MMS) and corn-soy-soybean (MSS) on soil nitrogen fixing bacteria abundance and community structure were analyzed by fluorescence quantitative PCR and high-throughput sequencing. The results showed that soil organic matter (SOM), total phosphorus (TP), available nitrogen (AN) and available phosphorus (AP) contents in rotation were significantly higher than those in continuous cropping. The abundance of nitrogen-fixing bacteria in rotation was significantly higher than that in CM and significantly lower than that in CS. The diversity of NITR Ogen-fixing bacteria in MMS and MSS was significantly higher than that in CM. Total nitrogen (TN) was the main driving factor for the change of nitrogen-fixing bacteria community structure. Planting system indirectly affected the abundance and diversity of nitrogen-fixing bacteria through soil chemical properties. These results indicate that MSS and MMS are more conducive to the propagation of soil nitrogen-fixing bacteria in the semi-arid area of Western Jilin province, which can provide scientific basis for rational planting and nitrogen regulation from the perspective of microbiology.
FENG Xue-wan , LI Cui-lan , PENG Chang , ZHANG Jin-jing , LIN Yuan , LIU Hang . Effects of corn-soybean rotation system on soil nitrogen-fixing bacteria community structure and quality in black soil[J]. JOURNAL OF NATURAL RESOURCES, 2022 , 37(9) : 2319 -2333 . DOI: 10.31497/zrzyxb.20220909
表1 土壤化学性质的测定方法Table 1 Determination methods of soil chemical properties |
化学性质 | 测定方法 |
---|---|
土壤有机质SOM | K2CrO7氧化—外加热法 |
pH | pH计土壤pH (水土比为2.5∶1) |
土壤全氮TN | 凯氏定氮法 |
土壤全磷TP | HClO4-H2SO4消解—钼锑抗比色法 |
土壤全钾TK | 火焰分光光度计法 |
土壤速效氮AN | 碱解扩散法 |
土壤速效磷AP | 钼锑抗比色法 |
土壤速效钾AK | 火焰分光光度计法 |
表2 荧光定量PCR的反应体系Table 2 Reaction system of fluorescence quantitative PCR |
成分 | 体积/μL |
---|---|
nifH-F (10 μM) | 1 |
nifH-R (10 μM) | 1 |
SYBR®PremixEx Taq™II (TliRNaseH Plus), ROX plus | 1 |
2×Taq Master Mix | 25 |
水 | 22 |
总共 | 50 |
表3 荧光定量PCR的扩增条件Table 3 Amplification conditions of fluorescence quantitative PCR |
程序 | 温度/℃ | 时间/秒 |
---|---|---|
预变性 | 94 | 300 |
变性 | 55 | 30 |
退火 | 2 | 30 |
延伸 | 72 | 600 |
表4 PCR的反应体系Table 4 Reaction systems of high-throughput sequencing |
成分 | 体积/μL |
---|---|
2×Taq Plus Master Mix | 12.5 |
BSA (2 ng/μl) | 3 |
Forward Primer (5 μM) | 1 |
Reverse Primer (5 μM) | 1 |
DNA (加入的DNA总量为30 ng) | 2 |
dd H2O | 5.5 |
总共 | 25 |
表5 PCR的反应参数Table 5 PCR reaction parameters |
程序 | 温度/℃ | 时间/秒 |
---|---|---|
预变性 | 95 | 300 |
变性 | 55 | 45 |
退火 | 72 | 50 |
表6 玉米—大豆轮作和连作下土壤的化学性质Table 6 Soil chemical properties under corn-soybean rotation and continuous cropping |
处理 | pH | SOM /(g/kg) | TN /(g/kg) | TP /(g/kg) | TK /(g/kg) | AN /(mg/kg) | AP /(mg/kg) | AK /(mg/kg) |
---|---|---|---|---|---|---|---|---|
CS | 6.97±0.06ab | 11.95±1.24c | 1.87±0.01b | 0.55±0..02c | 22.53±0.31c | 109.27±8.48b | 22.28.±0.39b | 156.17±3.01a |
CM | 6.90±0.09b | 12.99±0.18c | 1.80±0.04c | 0.56±0.02c | 25.53±1.03a | 117.08±2.42b | 22.28±2.68b | 163.00±4.82a |
MS | 7.15±0.22ab | 15.20±0.15b | 1.88±0.01b | 0.68±0.04a | 24.93±1.45ab | 132.03±11a | 44.13±7.69a | 162.50±15.52a |
MMS | 7.22±0.15a | 16.83±0.48a | 2.06±0.01a | 0.65±0.05ab | 24.73±0.23ab | 123.67±0.67ab | 36.62±2.61a | 168.33±5.97a |
MSS | 7.20±0.18a | 17.73±0.82a | 2.09±0.03a | 0.58±0.06bc | 23.60±1.20bc | 123.09±4.03ab | 36.62±8.97a | 159.50±1.80a |
注:同列不同小写字母表示不同处理间差异显著,下同。 |
表7 土壤固氮菌nifH基因 Illumina MiSeq 高通量测序数据(97%相似度)Table 7 Illumina MiSeq sequenced bacterial data (at 97% sequence similarity) based on the nifH gene |
处理 | 序列/条 | Final TAG OTU/个 |
---|---|---|
CS1 | 142036 | 499 |
CS2 | 90760 | 324 |
CS3 | 106553 | 324 |
CM1 | 141014 | 576 |
CM2 | 71625 | 526 |
CM3 | 135340 | 557 |
MS1 | 26825 | 412 |
MS2 | 76579 | 436 |
MS3 | 155086 | 535 |
MMS1 | 142022 | 532 |
MMS2 | 142081 | 486 |
MMS3 | 73582 | 576 |
MSS1 | 143927 | 523 |
MSS2 | 168971 | 548 |
MSS3 | 31076 | 470 |
图2 玉米—大豆轮作和连作条件下Proteobacteria的相对丰度和土壤固氮菌优势菌属水平上的群落组成注:*:P<0.05,ANOVA差异分析。颜色代表属相对丰度高低,粉色为低丰度,红色为高丰度。 Fig. 2 The relative abundance of Proteobacteria and the community composition at the level of the dominant genus of soil nitrogen-fixation bacteria under the conditions of corn-soybean rotation and continuous cropping |
图4 玉米—大豆轮作和连作条件下土壤固氮菌群落的包作非度量多维尺度分析及土壤固氮菌群落结构和环境因子的典范对应分析Fig. 4 Inclusion nonmetric multidimensional scale analysis (NMDS) of soil nitrogen-fixing bacteria community and canonical correspondence analysis (CCA) of soil nitrogen-fixing bacteria community structure and environmental factors under corn-soybean rotation and continuous cropping |
表8 玉米—大豆轮作和连作条件下土壤固氮菌群落结构和环境因子的典范对应分析的解释率和贡献率Table 8 Explanation rate and contribution rate of canonical correspondence analysis of soil nitrogen-fixing bacteria community structure and environmental factors under corn-soybean rotation and continuous cropping |
指标 | 解释率/% | 贡献率/% | P |
---|---|---|---|
SOM | 4.1 | 6.6 | 0.764 |
pH | 7.3 | 11.6 | 0.236 |
TN | 12.4 | 19.6 | 0.01 |
TP | 5.9 | 9.4 | 0.554 |
TK | 11.5 | 18.3 | — |
AN | 7.6 | 12.6 | 0.16 |
AP | 7.9 | 12.6 | 0.146 |
AK | 5.9 | 9.3 | — |
图7 玉米—大豆轮作和连作土壤固氮菌共线性网络分析注:节点颜色表示不同土壤固氮菌门,氮菌网络图红线表示两个节点之间的正相关,绿线表示负相关。 Fig. 7 Soil consolidation in corn-soybean rotation and continuous cropping |
表9 土壤固氮菌网络的拓扑特征Table 9 Topological characteristics of nitrogen-fixing bacteria network |
处理 | 连作 | 轮作 |
---|---|---|
节点/个 | 224 | 296 |
边/条 | 2530 | 693 |
平均度/(条/个) | 28.657 | 6.037 |
网络直径/链路树 | 6 | 12 |
平均路径长度/链路树 | 2.682 | 4.719 |
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