Spectral characteristics of soil dissolved organic matter in different rice-crayfish cultivation modes
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摘要: 稻虾种养的农业活动显著改变了田间生物化学因子, 土壤溶解性有机质(DOM)是其中之一。本研究利用紫外-可见吸收光和三维荧光光谱技术, 耦合平行因子分析, 对有环沟式稻虾轮作和共作、无环沟式稻虾共作和传统稻作4种稻虾种养模式土壤DOM的特征进行了研究。结果显示, 不同稻虾种养模式土壤DOM紫外吸光度和比吸收系数均低于传统稻作, 光谱斜率比(SR)均值为0.9~2.0; 254 nm和365 nm处紫外吸光度的比值(E2/E3)在无环沟稻虾共作模式中最小, 水稻单作中E2/E3值相对于稻虾种养模式大; 300 nm和400 nm处紫外吸光度的比值(E3/E4)在稻虾种养模式均<3.5, 水稻单作>3.5。4种模式的荧光指数>1.9, 腐殖化指数<4, 自生源指数为0.6~0.7。不同模式均解析出2个类蛋白质组分(类络氨酸、类蛋白物质)和2个类腐殖质组分(腐殖酸、类腐殖质), 与生物源密切相关的类络氨酸组分占比较高且与其他组分均呈负相关, 其余组分相互呈正相关。上述结果中, 紫外-可见吸收光谱参数表明稻虾种养降低了土壤DOM的腐殖化程度、芳香性和物质组成, 稻虾种养土壤腐殖质类DOM腐殖酸为主, 水稻单作以富里酸为主; 4种模式的土壤DOM既有内源也有外源, 其中无环沟稻虾共作模式土壤DOM分子量最大, 且有一定外源性特征, 说明土壤保肥能力较好, 在4种模式中综合效益最佳; 三维荧光光谱分析表明不同模式下土壤DOM的物质组成无明显差异, 研究认为这与农田管理措施的间接影响等有关, 占比较高的络氨酸组分来源不同于其他组分, 可能与微生物对DOM或其他物质的分解转化有关, 而腐殖酸、类蛋白物质和类腐殖质组分很可能有高度的同源性; 不同种养模式之间的淹水差异通过影响溶解性有机碳(DOC)和DOM的释放, 从而影响种养模式之间DOM光谱特征。
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关键词:
- 稻虾种养 /
- 溶解性有机质(DOM) /
- 平行因子分析 /
- 三维荧光光谱 /
- 紫外-可见吸收光谱
Abstract: Rice and crayfish co-cultivation is the largest cultivation mode in China and is an important part of ecological agriculture. However, in the process of cultivation, the activities changed the physical and chemical properties of field soils, which had a significant effect on the change in organic matter. Dissolved organic matter (DOM), one of the most active components of organic matter, is also the subject of modification, but there are few related studies on the characteristics of modified dissolved organic matter. Therefore, in this study, ultraviolet-visible (UV-Vis) absorption spectra and three-dimensional fluorescence spectra were used to calculate different spectral parameters combined with parallel factor analysis (PARAFAC) to study the characteristics of soil DOM under four rice and cultivation modes (integrated rice-crayfish rotation system with ring groove, RS1; integrated rice-crayfish system with ring groove, RS2; integrated rice-crayfish system, RS0; traditional rice monoculture, MR). The results revealed that there were weak shoulder peaks at 260–280 nm in the UV-Vis absorption spectrum of soil DOM in all modes, and the absorbance decreased with increasing wavelength and gradually approached zero. The UV-Vis absorption spectral parameters SUVA254 (specific absorption coefficient) and UV254 (UV absorbance at wavelength 254 nm) of soil DOM in different rice and crayfish cultivation modes (RS1, RS2 and RS0) were lower than those of traditional rice monoculture (MR). The E2/E3 (ratio of UV absorbance at 254 nm to 365 nm) value was smallest in RS0, and the SR (spectral slope ratio) values of the two co-cropping patterns (RS2 and RS0) were averaged from 0.5 to 2.0. For all modes, the mean value of fluorescence index >1.9, humification index <4, biological index =0.6−0.7. All rice and crayfish cultivation modes had resolved two protein-like components (C1, C3) and two humic-like components (C2, C4), C1 accounted for a higher proportion and was negatively correlated with other components, and the remaining components were positively correlated with each other. The principal component analysis identified four effective components, which were arranged according to the contribution rate: PC1 represented the concentration and aromaticity of DOM; PC2 represented the biological characteristics, molecular weight and degree of humification of DOM; PC3 represented the protein components and autogenic features in DOM; PC4 represented the molecular structure of DOM. The results showed that many unsaturated aromatic structure molecules existed in the soil DOM of all modes. Rice and crayfish cultivation reduced the degree of humification and aromaticity of soil DOM. The RS0 mode had the largest molecular weight of soil DOM and certain exogenous characteristics; the soil fertilizer retention ability was better, and the comprehensive benefit was the best among the three rice and crayfish farming modes. There was no significant difference in the material composition of soil DOM under different modes, which was related to the indirect effects of agricultural management practices. The source of C1 was different from that of other components, which may be related to the decomposition and transformation of DOM by microorganisms. Additionally, the difference in flooding was one of the main reasons for the difference in soil DOM characteristics; the soil releases DOM and DOC through flooding, which significantly affects the characteristics of soil DOM. In production practice, attention should be paid to the application of organic fertilizers, which can maintain soil fertility, increase DOM and microbial diversity, maintain soil fertility, and monitor the status of contaminants in soil and crops on farmlands. -
图 1 不同稻虾种养模式与传统水稻单作田间试验布局示意图
Figure 1. Schematic diagrams of the field trails of different integrated rice-crayfish cultivation patterns and traditional rice monoculture
图 2 不同稻虾种养模式土壤溶解性有机质紫外-可见光吸收光谱图
RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture.
Figure 2. UV-Vis absorption spectra of soil dissolved organic matter in different integrated rice-crayfish cultivation patterns
图 3 不同稻虾种养模式土壤溶解性有机质的紫外-可见吸收光谱相关参数[UV254、SUVA254、SR、α(355)、E2/E3、E3/E4]
各参数的具体解释见表1。不同小写字母表示不同模式在P<0.05水平差异显著; RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。The description of each parameter is shown in the table 1. Different lowercase letters indicate significant differences among different modes at P<0.05 level. RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture.
Figure 3. Parameters of UV-Vis absorption spectral [UV254, SUVA254, SR, α(355), E2/E3, E3/E4] of soil dissolved organic matter in different integrated rice crayfish cultivation patterns
图 4 平行因子解析得出的不同稻虾种养模式土壤可溶性有机质中4个荧光组分与对应的载荷
RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。 RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture.
Figure 4. Four fluorescent components and loadings in soil dissolved organic mater of different integrated rice-crayfish cultivation patterns analyzed by parallel factor analysis (PARAFAC)
图 5 不同稻虾种养模式土壤溶解性有机质的4个组分荧光强度贡献百分比
RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture.
Figure 5. Contribution percentages of fluorescence intensities of four components of soil dissolved organic matter in different integrated rice-crayfish cultivation patterns
图 6 不同稻虾种养模式土壤溶解性有机质的荧光光谱参数
不同小写字母表示不同模式在P<0.05水平差异显著; RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。FI: 荧光参数; BIX: 自生源指数; HIX: 腐殖化指数。Different lowercase letters indicate significant differences among different modes at P<0.05 level. RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture. FI: fluorescence index; BIX: biological indix; HIX: humification index.
Figure 6. Fluorescence parameters of soil dissolved organic matter in different integrated rice crayfish cultivation patterns
图 7 不同稻虾种养模式土壤溶解性有机质(DOC)组分与紫外-可见吸收光谱参数和荧光参数的相关性
* 、**和***分别表示P≤0.05、P≤0.01和P≤0.001水平显著相关性显著; 图中圆形直径越大颜色越深表示相关性越强。C1、C2、C3和C4分别为荧光组分类络氨酸、腐殖酸、类蛋白和类腐殖质, 其他的具体解释见表1。RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。*, ** and *** mean significant correlation at P≤0.05, P≤0.01 and P≤0.001 levels, respectively. The larger the circular diameter and the darker the color, the stronger the correlation. C1, C2, C3 and C4 are fluorescent components of tyrosine-like amino acids, humic acid, proteoid and humic-like substances. The description of other abbraviations are shown in the table 1. RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture.
Figure 7. Correlations between soil dissolved organic matter (DOC) components and UV-Vis absorption spectral parameters and fluorescence parameters in different integrated rice-crayfish cultivation patterns
图 8 土壤溶解性有机质(DOC)组分与紫外-可见吸收光谱参数(a、b)、荧光参数(c、d)主成分分析的载荷(a、c)与得分(b、d)
“→”指示对应参数的载荷坐标方向和大小。C1、C2、C3和C4分别为荧光组分类络氨酸、腐殖酸、类蛋白和类腐殖质, 其他的具体解释见表1。RS1: 有环沟式稻虾轮作; RS2: 有环沟式稻虾共作; RS0: 无环沟式稻虾共作; MR: 水稻单作。“→” indicates the direction and magnitude of load coordinates of corresponding parameters in PCA analysis. C1, C2, C3 and C4 are fluorescent components of tyrosine-like amino acids, humic acid, proteoid and humic-like substances. The description of other abbraviations are shown in the table 1. RS1: integrated rice-crayfish rotation system with ring groove; RS2: integrated rice-crayfish system with ring groove; RS0: integrated rice-crayfish system; MR: traditional rice monoculture.
Figure 8. Loadings plot (a,c) and sample scores (b,d) in PCA between soil dissolved organic matter (DOC) components, UV-Vis absorption spectral parameters (a, b) and fluorescence parameters (c, d)
表 1 紫外-可见光吸收光谱与荧光光谱参数描述
Table 1. Description of parameters of UV-Vis absorption spectrum and fluorescence spectrum
光谱参数名称
Spectral parameter name定义与计算方法
Definition and calculation method参数指示意义
Idicative meaningUV254 波长254 nm处的紫外吸光度
UV absorbance at wavelength 254 nm表征溶解性有机质(DOM)的浓度和芳香性[19]
To characterize the concentration and aromaticity of dissolved organic matter (DOM)吸收系数
Absorption coefficient (aλ)αλ=2.303×Aλ/r; Aλ为波长λ处的吸光度, r为光程路径(m)
αλ=2.303×Aλ/r; Aλ is the absorbance at wavelength λ, r is the optical path (m).表征样品中CDOM (有色可溶性有机质)相对浓度[20]
To characterize the relative concentration of colored DOM in samples.E2/E3 254 nm和365 nm处紫外吸光度的比值
Ratio of UV absorbance at 254 nm to 365 nm估算DOM分子量的相对大小, 与分子量呈反比[21]
To emstimat the relative size of DOM molecular, which inversely proportional to the molecular weightE3/E4 300 nm和400 nm处紫外吸光度的比值
Ratio of UV absorbance at 300 nm to 400 nm衡量腐殖质的腐殖化程度及其芳香性[21]
To characterizes the degree of humification and aromaticity of humus比吸收系数
Specific absorption coefficient (SUVA254)UV254与可溶性有机碳(DOC)浓度的比值
Ratio of UV254 to concentration of dissolved organic carbon (DOC)与DOM芳香度密切相关, 表征DOM的芳香性, 值越大, 芳香性越高[22]
Closely positively correlated to DOM aromaticity, and represent the aromaticity of DOM光谱斜率比
Spectral slope ratio (SR)275~295 nm波长和350~400 nm波长吸光度指数函数曲线光谱斜率的比值, 即
SR=S(275~295)/S(350~400)。
Ratio of the spectral slope of the absorbance index function curve for the wavelength of 275 nm to 295 nm and the wavelength of 350−400 nm, SR=S(275−295)/S(350−400)反映DOM的来源与分子量大小: 当SR>1, 表示以生物源为主; SR<1则为外源。SR越大表明DOM 分子量越小[23]
Reflect the source and molecular weight of DOM. When SR>1, the DOM is mainly from biological sources; while when SR<1, it is exogenous. The larger the value, the smaller the DOM molecular weight.荧光指数
Fluorescence index (FI)指激发波(Ex)在370 nm处, 发射波长(Em)在
450 nm与500 nm处的荧光强度比值
Refers to the fluorescence intensity ratio between excitation wavelength at 370 nm and emission wavelength at 450 nm and 500 nm反映芳香氨基酸和其他物质对荧光强度的贡献, 可作为来源指示。FI>1.9, 生物代谢和释放为主要来源, 自生源突出; FI<1.4, 植物和土壤有机物贡献为主, 外源性突出[24]
Reflect the contribution of aromatic amino acids and other substances to fluorescence intensity, it can be used as source indicator. If FI >1.9, the biological metabolism and release are the main sources, autogenesis is prominent. if FI<1.4, the plant and soil organic matter are the main source, showing exogenous featuer.腐殖化指数
Humification index (HIX)激发波长(Ex)在254 nm处, 发射波长(Em)在435~480 nm与300~345 nm之间的积分值比值
Ratio of the integral value between the excitation wavelength at 254 nm and the emission wavelength at 435−480 nm and 300−345 nm衡量DOM的腐殖化程度, 指数值越高, 腐殖化程度越高。其中, HIX>6表示高度腐殖化且陆源贡献较大; 4~6表示高度腐殖化具有微弱的自生源特性; HIX<4则表示腐殖化程度弱, 以自生源为主[25]
Measure the degree of humification of DOM. The higher the index value, the higher the degree of humification. HIX>6 indicates high humification with large terrestrial contribution. 4−6 HIX indicates high humification with weak autobiographical characteristics. HIX<4 indicates humification degree is weak and autogenous.自生源指数
Biologcal index (BIX)指激发波长(Ex)在310 nm处, 发射波长(Em)在380 nm与430 nm处的荧光强度比值
Fluorescence intensity ratio between excitation wavelength at 310 nm and emission wavelength at 380 nm and 430 nm.反映DOM中新鲜产生的物质比例, 可评估DOM所在环境中的生物活性程度。BIX>1为生物源, BIX<0.7则表示低本土成分, 0.7~0.8为中等本土成分, 0.8~1.0为强本土成分[26]
Reflect the proportion of freshly produced substances in DOM, the degree of biological activity in the environment. BIX>1 is biogenic, BIX<0.7 is low native component, 0.7−0.8 is medium native component, and 0.8−1.0 is strong native component.
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