Mechanism of Interaction Between Calcium Ion and Hydrogen Sulfide Alleviating the Inhibitory Effect of Aluminum on Root Elongation in Rice

doi:10.16819/j.1001-7216.2023.220603

Abstract

Abstract:

【Objective】 The objective of this study is to elucidate the molecular and physiological mechanisms of the interaction between calcium ion (Ca²⁺) and hydrogen sulfide (H₂S) alleviating aluminum (Al) toxicity in rice.【Method】 The rice cultivar kasalath was selected as test material, and treated with 0 μmol/L and 30 μmol/L AlCl₃, 0.1 mmol/L and 0.5 mmol/L CaCl₂, 0.2 μmol/L NaHS and 100 μmol/L H₂S scavenger hypotaurine (HP). The seeds were incubated at 30℃ in the dark for 24 h, then the root elongation, total Al content, Al content in cell sap, Al content in apoplast, Al content in cell wall, pectin content, pectin methylesterase activity and the relative expression of OsSTAR2, OsNRAT1 and OsFRDL4 were measured to explore the molecular and physiological mechanism of the interaction between Ca²⁺ and H₂S alleviating Al toxicity in rice. 【Result】 Under Al stress, compared with 0.1 mmol/L CaCl₂ treatment, 0.5 mmol/L CaCl₂ treatment significantly increased root elongation, H₂S content, total calcium (Ca) content and cytoplasm Ca content, and significantly decreased total Al content in root, cell sap Al content, apoplastic Al content and cell wall Al content in rice. The pretreatment of NaHS significantly increased the elongation of rice roots at the two calcium concentrations, accompanied with the significant increase of root tip Al content, root total Al content, cell sap Al content, apoplastic Al content and cell wall Al content. The relative expression levels of OsSTAR2 and OsFRDL4 were significantly increased and the relative expression of OsNRAT1 was significantly decreased at the two Ca²⁺ concentrations after pretreated with NaHS. However, the application of HP produced an opposite result. 【Conclusion】 Under Al stress, Ca²⁺ reduced the accumulation of Al in rice root cell wall and the absorption of Al by rice roots through increasing the production of H₂S in rice root, and finally alleviated the inhibition of Al on rice root elongation.

Key words: rice, aluminum stress, calcium ion, hydrogen sulfide

摘要：

【目的】阐明钙离子与硫化氢相互作用缓解水稻铝毒害的分子和生理机制。【方法】以Kasalath为试验材料，选取0 μmol/L和30 μmol/L AlCl₃，0.1 mmol/L 和 0.5 mmol/L CaCl₂，0.2 μmol/L NaHS和100 μmol/L硫化氢清除剂亚牛磺酸(HP)作为处理浓度，将种子置于30℃培养箱中黑暗培养24 h后取水稻根系，通过测定水稻根系伸长量、总铝含量、细胞汁液中铝含量、质外体中铝含量、细胞壁中铝含量、果胶含量、果胶甲酯酶活性以及OsSTAR2、OsNRAT1和OsFRDL4相对表达量，探究钙离子与硫化氢互作缓解铝对水稻根系伸长抑制作用的机制。【结果】铝胁迫下，相较于0.1 mmol/L CaCl₂处理，0.5 mmol/L CaCl₂处理显著提高了水稻根系伸长量、硫化氢含量、总钙含量和细胞质中钙含量，显著降低了水稻根系的总铝含量，细胞液、质外体和细胞壁中的铝含量。铝胁迫下，硫氢化钠预处理后，水稻根系的伸长量在两种钙浓度下均显著增加，水稻的根尖铝含量、根系总铝含量、细胞液中铝含量、质外体中铝含量和细胞壁中铝含量在两种钙浓度下均显著降低，OsSTAR2和OsFRDL4相对表达量在两种钙浓度下均显著提高，OsNRAT1相对表达量在两种钙浓度下均显著降低。铝胁迫下，添加HP则呈现相反的结果。【结论】铝胁迫下，钙离子通过增加水稻根系硫化氢的生成，降低水稻根系对铝的吸收和积累，最终缓解铝对水稻根系伸长的抑制作用。

关键词: 水稻, 铝胁迫, 钙离子, 硫化氢

WEI Qianqian, XU Qingshan, PAN Lin, KONG Yali, ZHU Lianfeng, CAO Xiaochuang, TIAN Wenhao, LIU Jia, JIN Qianyu, XIANG Xingjia, ZHANG Junhua, ZHU Chunquan. Mechanism of Interaction Between Calcium Ion and Hydrogen Sulfide Alleviating the Inhibitory Effect of Aluminum on Root Elongation in Rice[J]. Chinese Journal OF Rice Science, 2023, 37(2): 142-152.

魏倩倩, 徐青山, 潘林, 孔亚丽, 朱练峰, 曹小闯, 田文昊, 刘佳, 金千瑜, 项兴佳, 张均华, 朱春权. 钙离子与硫化氢互作缓解铝对水稻根系伸长抑制作用的机制[J]. 中国水稻科学, 2023, 37(2): 142-152.

Figures/Tables 9

Table 1. Primers selected for this study and the sequences.

引物 Primer	序列Sequence (5′-3′)
OsSTAR2-R	CCTCAGCTTCTTCATCGTCACC
OsSTAR2-F	ACCTCTTCATGGTCACCGTCG
OsFRDL4-R	TCATTTGCGAAGAAACTTCCACG
OsFRDL4-F	CGTCATCAGCACCATCCACAG
OsNRAT1-F	GAGGCCGTCTGCAGGAGAGG
OsNRAT1-R	GGAAGTATCTGCAAGCAGCTCTGATGC
OsHistone-R	AACCGCAAAATCCAAAGAACG
OsHistone-F	GGTCAACTTGTTGATTCCCCTCT

Fig. 1. Effects of different Al3+ concentrations on root elongation (A) and Al content in root apices (B). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3).

Fig. 2. Effects of different Ca2+ concentrations on root elongation (A) and Al content in root apices (B). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). -Al, No Al3+ addition; +Al, 30μmol/L Al3+.

Fig. 3. Effects of different Ca2+ concentrations on total Al concentration (A), Al concentration in cell sap (B), apoplastic Al concentration (C), Al content in cell wall(D), pectin content (E), pectin methylesterase activity (F). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). 0.1Ca, 0.1 mmol/L CaCl2; 0.5Ca, 0.5 mmol/L CaCl2; 0.1Ca+Al, 0.1 mmol/L CaCl2+30 μmol/L AlCl3; 0.5Ca+Al, 0.5 mmol/L CaCl2+30 μmol/L AlCl3.

Fig. 4. Effects of different Ca2+ concentrations on total Ca concentration (A), Ca concentration in cell sap (B and C). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). 0.1Ca: 0.1 mmol/L CaCl2; 0.5Ca: 0.5 mmol/L CaCl2; 0.1Ca+Al: 0.1 mmol/L CaCl2+30 μmol/L AlCl3; 0.5Ca+Al: 0.5 mmol/L CaCl2+30 μmol/L AlCl3.

Fig. 5. Effects of different Ca2+ concentrations on the content of endogenous H2S. Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). 0.1Ca: 0.1 mmol/L CaCl2; 0.5Ca: 0.5 mmol/L CaCl2; 0.1Ca+Al: 0.1 mmol/L CaCl2+30 μmol/L AlCl3; 0.5Ca+Al: 0.5 mmol/L CaCl2+30 μmol/L AlCl3.

Fig. 6. Effects of interaction between Ca2+ and H2S on root elongation (A), relative root elongation (B) and Al content in root apices (C). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). 0.1Ca, 0.1 mmol/L CaCl2; 0.1Ca+NaHS, 0.1 mmol/L CaCl2+0.2 μmol/L NaHS; 0.1Ca+HP, 0.1 mmol/L CaCl2+100 μmol/L HP; 0.5Ca, 0.5 mmol/L CaCl2; 0.5Ca+NaHS, 0.5 mmol/L CaCl2+0.2 μmol/L NaHS; 0.5Ca+HP, 0.5 mmol/L CaCl2+100 μmol/L HP.

Fig. 7. Effects of interaction between Ca2+ and H2S on total Al concentration (A), Al concentration in cell sap (B), apoplastic Al concentration (C), Al content in cell wall(D), pectin content (E), pectin methylesterase activity (F). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). 0.1Ca, 0.1 mmol/L CaCl2; 0.1Ca+NaHS, 0.1 mmol/L CaCl2+0.2 μmol/L NaHS; 0.1Ca+HP, 0.1 mmol/L CaCl2+100 μmol/L HP; 0.5Ca, 0.5 mmol/L CaCl2; 0.5Ca+NaHS, 0.5 mmol/L CaCl2+0.2 μmol/L NaHS; 0.5Ca+HP, 0.5 mmol/L CaCl2+100 μmol/L HP.

Fig. 8. Effects of interaction between Ca2+ and H2S on relative expression of OsSTAR2 (A), OsFRDL4 (B) and OsNRAT1 (C). Different letters in the figure indicate that there is a significant difference in the results under the analysis of variance (P<0.05), and the value is the mean ± standard deviation (n=3). 0.1Ca, 0.1 mmol/L CaCl2; 0.1Ca+NaHS, 0.1 mmol/L CaCl2+0.2 μmol/L NaHS; 0.1Ca+HP, 0.1 mmol/L CaCl2+100 μmol/L HP; 0.5Ca, 0.5 mmol/L CaCl2; 0.5Ca+NaHS, 0.5 mmol/L CaCl2+0.2 μmol/L NaHS; 0.5Ca+HP, 0.5 mmol/L CaCl2+100 μmol/L HP.

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