Effects of Straw-returning on Phosphorus Morphology and Microbial Phosphorus-cycling Genes in Rice Paddy Soil
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摘要:
目的 以南方酸性土壤连续8年秸秆还田定位试验田为研究对象，探究秸秆还田对水稻分蘖期与成熟期土壤磷循环微生物功能基因的影响。 方法 定位试验共设置5个处理：化肥+无秸秆还田（CK）、化肥+当季秸秆全量还田（CKS）、化肥+当季秸秆全量还田+秸秆替代10%钾肥（S10）、化肥+当季秸秆全量还田+秸秆替代20%钾肥（S20）和化肥+当季秸秆全量还田+秸秆替代30%钾肥（S30）。利用Guppy土壤磷素连续浸提法和宏基因组测序技术，分别测定土壤磷素组分和磷循环微生物功能基因数据。 结果 秸秆还田处理显著提高了Guppy土壤磷分级中可利用磷组分NaHCO3-Pi的含量（P＜0.05），其中S10、S20处理较CK处理提升了5.88%～8.73%；中等可利用磷组分中NaOH-Pi为南方酸性稻田土壤最主要的磷素形态，含量为154.03～202.11 mg·kg−1。同时，秸秆还田还显著影响了土壤磷循环微生物功能基因的丰度，其中无机磷溶解基因pqqC对秸秆全量还田处理的响应更为明显。秸秆还田条件下磷循环功能基因（如phnW、phnO、pqqB、pqqC）促进了盐酸磷和残余磷向可利用磷库的活化，appA、phnX、ppx基因参与了稳定态磷的活化过程，NaOH-Pi为关键的过渡态磷素，参与长期的磷素转化过程。此外，土壤有机碳和pH是决定功能基因丰度的主要影响因子。 结论 秸秆还田通过影响土壤理化性质，改变了稻田土壤磷循环功能基因丰度，促进了南方酸性稻田土壤磷素转化过程。 Abstract:Objective Effects of straw-returning on phosphorus morphology and microbial phosphorus-cycling genes in paddy soil at rice tillering and maturing stages were investigated. Method In consecutive 7 years on a rice field in southern China under a positioning experiment, spent straws were returned to the acidic soil. The implemented treatments included: (1) chemical fertilizer without straw-returning (CK), (2) chemical fertilizer + 100% straw-returning in same season (CKS), (3) CKS+ straws to replace 10% potassium fertilizer (S10), (4) CKS + straws to replace 20% potassium fertilizer (S20) or (5) CKS + straws to replace 30% potassium fertilizer (S30). At end of the treatments, Guppy soil phosphorus continuous extraction method and metagenomic technology were applied to determine the composition of phosphorus of different forms and microbial phosphorus-cycling genes. Result Straw-returning significantly increased the available NaHCO3-Pi in soil (P＜0.05)—the S10 and S20 treatments resulted in an increase by 5.88%-8.73% over CK. NaOH-Pi was the main form of phosphorus in the acid paddy soil in southern China with a content ranging from 154.03 mg·kg−1 to 202.11 mg·kg−1. By turning the spent straws into the field, the abundance of phosphorus-cycling genes, especially the inorganic phosphorus dissolution gene pqqC under CKS, was significantly affected. The genes, such as phnW, phnO, pqqB, and pqqC, activated the conversion of hydrochloric acid phosphorus and residual phosphorus into available form; those like appA, phnX, and ppx, participated in the formation of stable phosphorus; and NaOH-Pi played a key role in the long-term transformation of the mineral. And the main factors that governed the abundance of the functional genes appeared to be the organic carbon and pH of the soil. Conclusion Through altering the soil physiochemical properties, returning spent straws to the ground significantly enriched the microbial phosphorus-cycling genes that promoted the mineral transformation of the acidic paddy soil in southern China. -
Key words:
- Straw-returning /
- phosphorus forms /
- phosphorus-cycling genes /
- metagenomics
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图 1 水稻不同生育期不同处理土壤可利用磷组分及含量
图中不同小写字母表示处理间差异显著（P＜0.05）。下同。
Figure 1. Easily available phosphorus in soil under various treatments at rice growth stages
Data with different lowercase letters indicate significant differences at P＜0.05. Same for below.
图 2 水稻不同生育期不同处理土壤中等可利用磷组分及含量
Figure 2. Moderately available phosphorus in soil under various treatments at rice growth stages
图 3 水稻不同生育期不同处理土壤稳定态磷组分及含量
Figure 3. Non-available phosphorus in soil under various treatments at rice growth stages
图 4 土壤有机磷矿化基因的相对丰度
A和B分别代表水稻分蘖期与成熟期编码酸性磷酸酶、碱性磷酸酶和植酸酶的基因，C和D分别表示水稻分蘖期与成熟期编码C-P键裂解酶的基因。*（ P ＜0.05），**（ P ＜0.01）。
Figure 4. Relative abundance of organic phosphorus-mineralizing genes in soil
A and B represent the genes encoding acid phosphatase, alkaline phosphatase and phytase at tillering and maturing stages, C and D represent the genes encoding C-P bond cleavage enzymes at tillering and maturing stages, respectively. * (P ＜0.05), ** (P ＜0.01).
图 5 土壤无机磷溶解基因的相对丰度
A和B分别表示水稻分蘖期和成熟期，下同。
Figure 5. Relative abundance of organic phosphorus-mineralizing genes in soil
A and B represent tillering and maturing stages, same for below.
图 6 土壤理化性质与磷循环功能基因的相关性分析
Figure 6. Correlation between physiochemical properties and phosphorus-cycling genes in soil
图 7 土壤磷素组分与磷循环功能基因的相关性分析
Figure 7. Correlation between phosphorus components and phosphorus-cycling genes in soil
表 1 不同处理化肥养分投入量
Table 1. Nutrients in treatment fertilizers
（kg·hm−2） 处理
Treatment早稻 Early rice 晚稻 Late rice 总计 Total N P2O5 K2O N P2O5 K2O N P2O5 K2O CK 155.3 47.3 135.0 155.3 47.3 135.0 310.5 94.5 270.0 CKS 155.3 47.3 135.0 155.3 47.3 135.0 310.5 94.5 270.0 S10 155.3 47.3 121.5 155.3 47.3 121.5 310.5 94.5 243.0 S20 155.3 47.3 108.0 155.3 47.3 108.0 310.5 94.5 216.0 S30 155.3 47.3 94.5 155.3 47.3 94.5 310.5 94.5 189.0 
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表 2 磷循环微生物功能基因
Table 2. Microbial phosphorus-cycling genes
功能基因分组
Functional gene grouping基因名称
Gene nameKEGG编号
KEGG number基因功能详情
Gene Function Detials有机磷矿化基因
Genes involved in organic P-mineralizationphoN K09474 酸性磷酸酶 （A） Acid phosphatase （class A） olpA K01078 酸性磷酸酶（C）acid phosphatase （class C） aphA K03788 酸性磷酸酶 （B） Acid phosphatase （class B） phoA K01077 碱性磷酸酶 Alkaline phosphatase phoD K01113 碱性磷酸酶 Alkaline phosphatase D phoX K02040 碱性磷酸酶 Alkaline phosphatase phyA K01083 3-植酸酶 3-phytase appA K01093 4-植酸酶/酸性磷酸酶 4-phytase/acid phosphatase phnW K03430 2-氨基乙基膦酸-丙酮酸转氨酶 2-aminoethylphosphonate-pyruvate transaminase phnX K05306 膦酸乙醛水解酶 Phosphonoacetaldehyde hydrolase phnA K06193 烷基膦酸酯利用操纵子 Alkylphosphonate utilization operon protein phnN K05774 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnL K05780 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnM K06162 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnF K02043 酸酯转运系统调控蛋白 Phosphonate transport system regulatory protein phnG K06166 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnH K06165 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnI K06164 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnJ K06163 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnO K09994 碳-磷键裂解酶复合物 C-P lyase multienzyme complex phnK K05781 C-P 裂解酶亚基，α-d-核糖 1-甲基膦酸 5-三磷酸合酶
C-P lyase subunit，alpha-D-ribose 1-methylphosphonate 5-triphosphate synthasephnP K06167 酸核糖基1,2-环磷酸二酯酶 Phosphoribosyl 1,2-cyclicphosphate phosphoDiesterase 无机磷溶解基因
Inorganic phosphate solubilizationppa K01507 无机焦磷酸酶 Inorganic pyrophosphatase ppx K01524 外切多聚磷酸酶 Exopolyphosphatase/鸟苷-5'-三磷酸，3'-二磷酸焦磷酸酶
Guanosine-5'-triphosphate 3'-diphosphate pyrophosphataseppk K00937 多聚磷酸激酶 Polyphosphate kinase gcd K00117 葡萄糖脱氢酶 Quinoprotein glucose dehydrogenase pqqB K06136 吡咯喹啉醌合成蛋白 B Pyrroloquinoline quinonebiosynthesis protein B pqqC K06137 吡咯喹啉醌合成酶 Pyrroloquinoline-quinone synthase pqqD K06138 吡咯喹啉醌合成蛋白 Pyrroloquinoline quinonebiosynthesis protein pqqE K06139 PqqA 肽环酶 PqqA peptide cyclase
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