首页分享硝态氮抑制水稻铁向上运输加剧低镉胁迫的机理

硝态氮抑制水稻铁向上运输加剧低镉胁迫的机理

来源：花匠小妙招时间：2025-05-22 20:44

摘要:

目的

铵态氮减少作物对镉(Cd)的吸收，而硝态氮则增加对Cd的吸收。通过研究供应两种形态氮对水稻镉吸收的影响及其作用机理，为水稻氮肥管理降镉策略提供理论依据。

方法

以粳稻品种‘中花11号’(Oryza sativa L.，ZH11) 为材料进行了水培试验。在Hoagland营养液基础上，分别设置硝态氮、铵态氮浓度为1.25 mol/L的营养液，每个营养液中设置添加和不添加Cd 0.5 μmol/L，共4个处理。水稻幼苗在处理液中生长21天后，取样调查水稻生长指标，地上部和根部铁(Fe)、锌(Zn)、Cd含量, 根部亚细胞组分Fe、Cd含量和金属离子转运相关基因表达量，结合结构方程模型分析各生理指标间的相互作用关系。

结果

铵态氮条件下低Cd胁迫对水稻生长无显著影响，而硝态氮条件下低Cd胁迫显著降低了水稻叶片SPAD、株高、地上部及根部干物质量，降幅分别为46%、27%、36%、25%，且叶片出现缺绿症状。叶片SPAD值与地上部Fe含量正相关 (R2=0.79)，与Cd含量负相关，与Zn含量无关，说明硝态氮下叶片SPAD值下降是由Fe含量降低、Cd含量升高所致。低Cd胁迫下，硝态氮处理水稻根表、根内Fe含量分别高于、低于铵态氮处理，因而根表Fe向根内的转移系数低于铵态氮；亚细胞Fe含量分析结果表明，硝态氮处理根系细胞壁、细胞器Fe含量高于铵态氮处理，而可溶性部分Fe含量低于铵态氮处理，因而Fe的移动性较差，Fe从根部向地上部的转移系数低。同时，硝态氮处理提高了根部Fe吸收运转基因OsIRT1、OsIRT2的表达。

结论

低Cd胁迫下，供应硝态氮显著降低水稻叶片SPAD值，抑制根及地上部生长，并因缺Fe导致叶片失绿，而供应铵态氮不会产生缺Fe现象。硝态氮促进了水稻根表Fe膜的形成及Fe在细胞壁的富集，降低了Fe的运转系数，致使地上部Fe供应不足，植物体内缺Fe信号反馈通过上调Fe吸收转运调控相关基因的表达，相关转运蛋白的非特异性吸收导致Cd累积量显著增加，抑制了水稻的生长发育。

关键词: 水稻 / 硝态氮 / 镉 / 铁吸收转运

Abstract:

Objectives

Ammonium nitrogen (NH4+-N) reduces the uptake of cadmium (Cd) by crops, whereas nitrate nitrogen (NO3−-N) increases Cd uptake. Research on the effects of supplying these two nitrogen forms on cadmium absorption in rice and the underlying mechanisms provides a theoretical basis for developing cadmium-reducing strategies in nitrogen fertilizer management for rice cultivation.

Methods

A hydroponic experiment was conducted using the japonica rice variety ‘Zhonghua 11’ (Oryza sativa L., ZH11). Based on Hoagland nutrient solution, we set up nutrient solutions with nitrate nitrogen and ammonium nitrogen at a concentration of 1.25 mol/L, including treatments with and without 0.5 μmol/L Cd, resulting in four treatments overall. After 21 days of growth in the treatment solution, we sampled to investigate rice growth indicators, the contents of iron (Fe), zinc (Zn), and Cd in the above-ground and root parts, subcellular Fe and Cd contents in roots, and the expression of genes related to metal ion transport. We analyzed the interactions among various physiological indicators using structural equation modeling.

Results

Under ammonium nitrogen conditions, low cadmium stress had no significant effect on rice growth. However, under nitrate nitrogen conditions, low cadmium stress significantly reduced leaf SPAD values, plant height, and dry biomass of both the above-ground and root parts by 46%, 27%, 36%, and 25%, respectively, and the leaves showed symptoms of chlorosis. The SPAD value of the leaves was positively correlated with the Fe content in the above-ground part (R2=0.79), negatively correlated with Cd content, and had no relation to Zn, indicating that the decrease in SPAD values under nitrate nitrogen was due to reduced Fe content and increased Cd content. Under low Cd stress, the Fe content in root surface and internal roots of rice treated with nitrate nitrogen was higher and lower than that treated with ammonium nitrogen, respectively, resulting in a lower transfer coefficient of Fe from the root surface to the inner root compared to ammonium nitrogen. Subcellular Fe content analysis showed that the Fe content in the root cell walls and organelles of nitrate nitrogen treatment was higher than that of ammonium nitrogen treatment, while the Fe content in soluble parts was lower than that of ammonium nitrogen, leading to poorer mobility of Fe and a lower transfer coefficient of Fe from roots to above-ground parts. Simultaneously, nitrate nitrogen treatment enhanced the expression of root Fe uptake transport genes OsIRT1 and OsIRT2.

Conclusions

Under low Cd stress, supplying nitrate nitrogen significantly decreased rice leaf SPAD values, inhibited growth of the roots and above-ground parts, and caused leaf chlorosis due to Fe deficiency. In contrast, supplying ammonium nitrogen did not result in Fe deficiency. Nitrate nitrogen promoted the formation of Fe membranes at the root surface and the accumulation of Fe in the cell walls, reducing the transfer coefficient of Fe and resulting in insufficient Fe supply in the above-ground parts. The feedback of Fe deficiency signals within the plant upregulated the expression of genes regulating Fe absorption and transport, and the nonspecific absorption of related transport proteins led to a significant increase in Cd accumulation, inhibiting rice growth and development.

图 1 低镉胁迫下不同氮供应形态对水稻生长的影响

注：CK和Cd分别表示营养液中不添加和添加0.5 μmol/L CdCl2。柱上不同小写字母表示处理间差异显著 (P<0.05)。

Figure 1. Effects of different nitrogen supply forms on rice growth under low cadmium stress

Note: CK, and Cd represent treatments without, and with addition of 0.5 μmol/L CdCl2. Different lowercase letters above the bars indicate significant difference among treatments (P<0.05).

图 2 氮形态与低镉胁迫处理对水稻地上部和根部金属离子含量的影响

注：CK、Cd分别表示营养液中不添加、添加0.5 μmol/L CdCl2，NH4+、NO3−代表营养液中氮供应分别为1.25 mol/L (NH4)2SO4、Ca(NO3)2。柱上不同小写字母表示处理间差异显著 (P<0.05)。

Figure 2. Effects of nitrogen forms and low Cd stress on element content in aboveground and root of rice

Note: CK, and Cd represents treatments without, and with addition of 0.5 μmol/L CdCl2. NH4+, and NO3− indicate treatments of supplying 1.25 mol/L (NH4)2SO4, and Ca(NO3)2, respectively. Different lowercase letters above the bars indicate significant difference among treatments (P<0.05).

图 3 水稻金属含量与SPAD水平相关性分析

注：Cd表示添加0.5 μmol/L CdCl2的低镉处理，NH4+、NO3−代表营养液中氮供应分别为1.25 mol/L (NH4)2SO4、Ca(NO3)2。

Figure 3. Analysis of the correlation between metal content and SPAD levels

Note: Cd indicates a low cadmium treatment with the addition of 0.5 μmol/L CdCl2; NH4+, and NO3− indicate treatments of supplying 1.25 mol/L (NH4)2SO4, and Ca(NO3)2, respectively.

图 4 不同氮形态对低镉处理水稻根部铁、镉吸收的影响

注：NH4+、NO3−代表营养液中氮供应分别为1.25 mol/L (NH4)2SO4、Ca(NO3)2。柱上*表示处理间差异达到0.05显著水平。

Figure 4. Effects of different nitrogen forms on iron and cadmium uptake in rice roots under low cadmium treatment

Note: NH4+, and NO3− indicate treatment of supplying 1.25 mol/L (NH4)2SO4, and Ca(NO3)2, respectively. The * above the bar indicate significant difference between treatments at 0.05 significant level.

图 5 氮形态对水稻根部亚细胞各组分铁镉累积量的影响

注：NH4+、NO3−代表营养液中氮供应分别为1.25 mol/L (NH4)2SO4、Ca(NO3)2。柱上*、**分别表示NH4+与NO3−处理间差异达到0.05、0.01显著水平。

Figure 5. Effects of nitrogen forms on iron and cadmium accumulation in subcellular fractions of rice roots

Note: NH4+, and NO3− indicate treatment of supplying 1.25 mol/L (NH4)2SO4, and Ca(NO3)2, respectively. The *, and ** above the bars indicate significant difference between NH4+、NO3− treatments at 0.05, and 0.01 significant levels, respectively.

图 6 不同氮形态对水稻根部铁转运调控基因表达量的影响

注：NH4+、NO3−代表营养液中氮供应分别为1.25 mol/L (NH4)2SO4、Ca(NO3)2。柱上*表示NH4+与NO3−处理间差异达到0.05显著水平。

Figure 6. Effects of different nitrogen forms on genes expression level in low cadmium treated rice roots

Note: NH4+, and NO3− indicate treatment of supplying 1.25 mol/L (NH4)2SO4, and Ca(NO3)2, respectively. The * above the bars indicate significant difference between NH4+ and NO3− treatments at 0.05 significant level.

图 7 结构方程模型

Figure 7. Structural equation modeling

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