Effects of different rice-crayfish models on community structure and diversity of sulfate-reducing bacteria
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摘要:
硫酸盐还原菌(SRB)作为一种土壤中普遍存在的微生物, 在生物地球化学循环中发挥关键作用。本研究以常规水稻单作模式为对照(CK), 无环沟式稻虾(水稻和克氏原螯虾)共作模式(RS0)、有环沟式稻虾轮作模式(RS1)与有环沟式稻虾共作模式(RS2)为研究对象, 采用16S rDNA高通量测序技术, 探究不同稻虾模式对土壤硫酸盐还原菌(SRB)群落结构及多样性的影响。结果表明: 与CK相比, 3种稻虾模式均显著降低了水稻成熟期土壤氧化还原电位(Eh)和硫化物、硫酸盐、全硫含量, 提高了pH和有机质、速效钾、碱解氮含量, 其中RS0提升效果最为显著; 3种稻虾模式SRB群落结构存在差异: RS1、RS2和RS0处理的δ-变形菌纲相对丰度比CK分别下降46.00%、63.61%和51.94%, 而α-变形菌纲则分别提高402.52%、441.01%和584.17%。3种稻虾模式硫酸盐还原菌Observed_species指数、Shannon指数和Simpson指数与CK相比均显著提高, 但不同稻虾模式间丰富度及多样性没有显著差异。RDA分析表明, 速效钾、Eh、有机质、有效磷、碱解氮、硫酸盐、硫化物、全硫和pH是影响稻田土壤SRB群落结构的影响因子, 其中全硫和硫化物是主要因子。综上, 稻虾模式能维持或者提高稻田土壤养分状况和SRB群落丰富度及多样性, 改善稻田土壤SRB群落结构, 结果可为稻虾种养土壤生态健康及稻田综合种养土壤微生物研究提供参考和支撑。
Abstract:The rice-crayfish integrated farming model, an ecological agricultural system, combines rice cultivation with the co-culture or rotation of crayfish (Procambarus clarkii), forming a mutually beneficial symbiosis. Sulfate-reducing bacteria (SRB) are widespread soil microbes that are pivotal in driving biogeochemical cycles. Researches on SRB is mainly centered on traditional rice fields, with comparatively few studies addressing their role in innovative rice-crayfish composite ecological agriculture systems. This study compared traditional rice monoculture (CK) with three distinct rice-crayfish systems: integrated rice-crayfish system without ring groove (RS0), integrated rice-crayfish rotation system with ring grooves (RS1), and integrated rice-crayfish system with ring grooves (RS2). Using 16S rDNA high-throughput sequencing technology, this study explored the effect of different rice-crayfish models on the community structure and diversity of soil SRB. Compared to the traditional rice monoculture, all three rice-crayfish models significantly decreased the soil oxidation-reduction potential (Eh) and contents of sulfide, sulfate, and total sulfur at the rice maturity stage. There was also a significantly increase in soil pH, and contents of organic matter, available potassium and nitrogen, with the most significant improvement observed in the RS0 model. This study revealed distinct SRB community structures across the three rice-crayfish models. Compared to the traditional rice monoculture, the δ-Proteobacteria class exhibited a decrease in relative abundance by 46.00%, 63.61%, and 51.94% in the RS1, RS2, and RS0 models, respectively, while the α-Proteobacteria class showed a substantial increase of 402.52%, 441.01%, and 584.17%, respectively. The Observed_species index, Shannon index, and Simpson index for SRB in all three rice-crayfish models were significantly higher than those in the CK, but there were no significant differences in richness and diversity among different rice-crayfish models. Redundancy analysis (RDA) revealed that factors such as available potassium, Eh, organic matter, available phosphorus, available nitrogen, sulfate, sulfide, total sulfur, and pH influenced the SRB community structure of rice field soil, with total sulfur and sulfide being the primary factors. In summary, the rice-crayfish model demonstrated the capacity to either maintain or improve the nutrient status, richness, and diversity of SRB communities in paddy soil, thereby effectively enhancing the structure of the SRB community in paddy fields. These findings offer valuable insights for supporting research on soil ecological health and soil microbiology of integrated rice-crayfish farming systems.
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Keywords:
- Rice-crayfish farming /
- Crop rotation /
- Co-culture /
- Sulfate reducting bacteria /
- Community structure /
- Diversity
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图 1 不同稻虾模式与传统水稻单作田间试验布局图
Figure 1. Layout of different rice-crayfish models and traditional rice in the field experiment
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图 2 不同稻虾模式和传统水稻单作土壤aprA基因维恩图
Figure 2. Microbial Venn diagram of aprA gene in soil samples of different rice-crayfish models and traditional monoculture of rice fields
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图 3 不同稻虾模式和传统水稻单作稻田土壤理化因子与硫酸盐还原菌群落Alpha多样性指数的相关性
Figure 3. Correlation between soil physicochemical factors and Alpha diversity index of soil sulfate-reducing bacteria communities in different rice-crayfish models and traditional monoculture rice fields
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图 4 不同稻虾模式和传统水稻单作土壤硫酸盐还原菌LEfSe分析
Figure 4. LAD Effect Size (LEfSe) analysis of sulfate-reducing bacteria in soil under different rice-crayfish and traditional monoculture rice fields
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图 5 稻田土壤理化因子与硫酸盐还原菌群落组成(门水平)的RDA分析
Figure 5. RDA analysis of paddy soil physicochemical factors and sulfate-reducing bacteria community composition at phylum level
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图 6 稻田土壤理化因子与硫酸盐还原菌群落优势属的相关性
Figure 6. Correlation of paddy soil physicochemical factors with dominant genuses of sulfate-reducing bacteria community
表 1 不同稻虾模式和传统水稻单作稻田土壤理化性质
Table 1 Physical and chemical properties of paddy soil under different rice-crayfish models and traditional monoculture of rice fields
处理
TreatmentpH 氧化还原电位
Oxidation-reduction
potential (mV)有效磷
Available P
(mg·kg−1)速效钾
Available K
(mg·kg−1)碱解氮
Available N
(mg·kg−1)有机质
Organic matter
(g·kg−1)硫化物
Sulfide
(mg·kg−1)硫酸盐
Sulfate
(mg·kg−1)全硫
Total sulfur
(g·kg−1)RS1 4.27±0.04c 440.57±6.20c 18.75±2.19bc 129.26±7.72b 117.95±2.28b 23.26±1.13b 0.31±0.02c 5273±220b 0.22±0.02b RS2 4.68±0.12b 448.30±5.41c 20.70±3.67b 129.99±1.36b 129.88±4.97a 22.97±3.50b 0.50±0.02b 4443±205c 0.16±0.02c RS0 4.82±0.03a 473.97±15.65b 27.35±0.60a 168.21±4.72a 126.72±3.13a 31.88±0.81a 0.52±0.01b 5303±211b 0.11±0.02d CK 4.09±0.02d 505.67±12.02a 15.18±0.55c 97.04±3.44c 102.79±6.39c 18.39±1.21c 1.82±0.09a 6533±311a 0.64±0.03a RS0、RS1、RS2和CK分别表示无环沟式稻虾共作模式、有环沟式稻虾轮作模式、有环沟式稻虾共作模式和传统水稻单作模式。表中同列不同小写字母表示处理间差异显著(P<0.05)。RS0, RS1, RS2 and CK represent the integrated rice-crayfish system without ring groove, integrated rice-crayfish rotation system with ring groove, integrated rice-crayfish system with ring groove, and traditional rice monoculture system, respectively. Different lowercase letters in the same column indicate significant differences among different treatments (P<0.05). 
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表 2 不同稻虾模式和传统水稻单作土壤硫酸盐还原菌群落的Alpha多样性指数
Table 2 Alpha diversity indexes of soil sulfate-reducing bacteria communities under different rice-crayfish models and traditional monoculture of rice fields
处理
Treatment覆盖率指数
Goods_coverage indexChao1指数
Chao1 indexObserved_species指数
Observed_species indexPD_whole_tree指数
PD_whole_tree indexShannon指数
Shannon indexSimpson指数
Simpson indexRS1 1.00±0.01a 4144.72±592.30a 3480.93±445.37a 286.30±159.62a 8.67±0.18a 0.99±0.00a RS2 1.00±0.01a 4088.99±585.93a 3487.67±321.10a 238.21±96.62a 8.84±0.25a 0.99±0.00a RS0 1.00±0.01a 3886.10±846.44a 3281.00±652.19a 288.95±184.42a 8.57±0.21a 0.99±0.01a CK 1.00±0.00a 2948.12±440.58a 2304.63±386.48b 155.84±70.34a 7.10±0.37b 0.97±0.01b RS0、RS1、RS2和CK分别表示无环沟式稻虾共作模式、有环沟式稻虾轮作模式、有环沟式稻虾共作模式和传统水稻单作模式。表中同列不同小写字母表示处理间差异显著(P<0.05)。RS0, RS1, RS2 and CK represent the integrated rice-crayfish system without ring groove, integrated rice-crayfish rotation system with ring groove, integrated rice-crayfish system with ring groove, and traditional rice monoculture system, respectively. Different lowercase letters in the same column indicate significant differences among different treatments (P<0.05).
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表 3 不同稻虾模式和传统水稻单作硫酸盐还原菌群落组成(纲水平)
Table 3 Composition of soil sulfate-reducing bacteria at class level in different rice-crayfish models and traditional monoculture of rice
菌纲 Class RS1 RS2 RS0 CK γ-变形菌纲 Gammaproteobacteria 33.05±1.90ab 35.40±4.24a 31.68±3.78ab 27.77±1.71b β-变形菌纲 Betaproteobacteria 20.60±0.81a 21.84±2.40a 21.11±2.32a 19.49±1.44a δ-变形菌纲 Deltaproteobacteria 13.37±0.74b 9.01±1.66b 11.90±3.82b 24.76±1.42a α-变形菌纲 Alphaproteobacteria 13.97±1.99a 15.04±0.91a 19.02±7.28a 2.78±0.64b 梭菌纲 Clostridia 4.62±0.73a 2.99±1.23b 3.00±0.71b 1.05±0.17c 绿菌纲 Chlorobia 1.88±1.13a 1.95±0.21a 2.28±1.31a 1.34±0.96a 其他纲 Others 0.04±0.01a ＜0.01d 0.02±0.01b ＜0.01c 未知纲 Unidentified 12.46±2.69b 13.76±2.73b 10.99±2.75b 22.81±1.99a RS0、RS1、RS2和CK分别表示无环沟式稻虾共作模式、有环沟式稻虾轮作模式、有环沟式稻虾共作模式和传统水稻单作模式。表中同行不同小写字母表示处理间差异显著(P＜0.05)。RS0, RS1, RS2 and CK represent the integrated rice-crayfish system without ring groove, integrated rice-crayfish rotation system with ring groove, integrated rice-crayfish system with ring groove, and traditional rice monoculture system, respectively. Different lowercase letters in the same line indicate significant differences among treatments (P<0.05).
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