中国水稻科学 ›› 2024, Vol. 38 ›› Issue (3): 290-302.DOI: 10.16819/j.1001-7216.2024.230910
魏倩倩1, 汪玉磊2, 孔海民2, 徐青山1, 颜玉莲1, 潘林1, 迟春欣1, 孔亚丽1, 田文昊1, 朱练峰1, 曹小闯1, 张均华1,*(), 朱春权1,*(
)
收稿日期:
2023-09-20
修回日期:
2023-11-22
出版日期:
2024-05-10
发布日期:
2024-05-13
通讯作者:
*email:zhangjunhua@caas.cn;
zhuchunquan@caas.cn
基金资助:
WEI Qianqian1, WANG Yulei2, KONG Haimin2, XU Qingshan1, YAN Yulian1, PAN Lin1, CHI Chunxin1, KONG Yali1, TIAN Wenhao1, ZHU Lianfeng1, CAO Xiaochuang1, ZHANG Junhua1,*(), ZHU Chunqun1,*(
)
Received:
2023-09-20
Revised:
2023-11-22
Online:
2024-05-10
Published:
2024-05-13
Contact:
*email:zhangjunhua@caas.cn;
zhuchunquan@caas.cn
摘要:
【目的】究基于施硫肥提高水稻体内硫化氢(H2S)生成缓解水稻在酸性土壤中铝(Al)毒害的栽培方式及其内在机制。【方法】以Al敏感品种Kasalath为试验材料,设置CK,CK+S(硫肥),CK+NaHS(H2S供体),Al,Al+S,Al+NaHS六个盆栽处理,通过测定不同时期水稻体内H2S含量、Al含量、抗氧化系统酶活性,分蘖期根系和光合作用相关指标,明确外源施加硫肥通过调控水稻体内H2S生成缓解Al毒的机制。 【结果】 与Al处理相比,Al+S处理显著提高水稻体内H2S含量,叶片中的H2S含量在分蘖期和齐穗期分别增加了19.27%和34.99%;根部H2S含量在分蘖期、齐穗期和成熟期分别增加了39.72%、21.08%和30.73%。相比Al处理,Al+S处理显著促进了分蘖期水稻根系的生长,提高了水稻的光合作用能力,同时降低了不同时期水稻根茎叶中的Al含量,在分蘖期分别下降了40.98%、28.47%和24.18%;齐穗期下降了14.58%、50.30%和13.17%;成熟期下降了14.44%、29.78%和8.70%。进一步研究发现,Al+S处理显著增强了水稻体内抗氧化酶(SOD、CAT、APX、POD)的活性,降低了水稻体内过氧化氢(H2O2)、丙二醛(MDA)、超氧阴离子(O2−·)含量,增加了与细胞质Al在液泡中进行分室相关的基因OsALS1以及与修复被Al破坏的细胞壁相关基因OsSTAR1和OsSTAR2的相对表达量。同时,外源施加硫肥还通过调控土壤pH值降低水稻根际土中交换态Al、无机吸附态Al和水溶态Al的含量,增加Al的水合物和氢氧化物的含量。Al+NaHS处理具有同样的效果。【结论】 外源增施硫肥能够通过提高水稻体内的H2S含量缓解Al对水稻根系生长和光合作用的抑制,提高抗氧化系统酶活性降低Al对水稻的过氧化损伤,同时促进液泡对水稻细胞质Al的区室化作用,降低细胞壁对Al的吸附或改变土壤中Al的形态,最终降低水稻根系内的Al含量来缓解Al对水稻生长的抑制作用。
魏倩倩, 汪玉磊, 孔海民, 徐青山, 颜玉莲, 潘林, 迟春欣, 孔亚丽, 田文昊, 朱练峰, 曹小闯, 张均华, 朱春权. 信号分子硫化氢参与硫肥缓解铝对水稻生长抑制作用的机制[J]. 中国水稻科学, 2024, 38(3): 290-302.
WEI Qianqian, WANG Yulei, KONG Haimin, XU Qingshan, YAN Yulian, PAN Lin, CHI Chunxin, KONG Yali, TIAN Wenhao, ZHU Lianfeng, CAO Xiaochuang, ZHANG Junhua, ZHU Chunqun. Mechanism of Hydrogen Sulfide, a Signaling Molecule Involved in Reducing the Inhibitory Effect of Aluminum Toxicity on Rice Growth Together with Sulfur Fertilizer[J]. Chinese Journal OF Rice Science, 2024, 38(3): 290-302.
基因Gene | 序列Sequence (5′-3′) |
---|---|
OsHistone-R | AACCGCAAAATCCAAAGAACG |
OsHistone-F | GGTCAACTTGTTGATTCCCCTCT |
OsALS1-F | GGTCGTCAGTCTCTGCCTTCTCO |
OsALS1-R | CCTCCCCATCATTTTCATTTGT |
OsSTAR1-F | TCGCATTGGCTCGCACCCT |
OsSTAR1-R | TCGTCTTCTTCAGCCGCACGAT |
OsSTAR2-F | ACCTCTTCATGGTCACCGTCG |
OsSTAR2-R | CCTCAGCTTCTTCATCGTCACC |
表1 本研究所选的引物及其序列
Table 1. Primers selected for this study and their sequences
基因Gene | 序列Sequence (5′-3′) |
---|---|
OsHistone-R | AACCGCAAAATCCAAAGAACG |
OsHistone-F | GGTCAACTTGTTGATTCCCCTCT |
OsALS1-F | GGTCGTCAGTCTCTGCCTTCTCO |
OsALS1-R | CCTCCCCATCATTTTCATTTGT |
OsSTAR1-F | TCGCATTGGCTCGCACCCT |
OsSTAR1-R | TCGTCTTCTTCAGCCGCACGAT |
OsSTAR2-F | ACCTCTTCATGGTCACCGTCG |
OsSTAR2-R | CCTCAGCTTCTTCATCGTCACC |
图1 不同处理下水稻体内H2S含量的变化图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。TS:分蘖期;HS:齐穗期;MS:成熟期。CK:0.12 g/kg KCl;CK+S: 0.12 g/kg K2SO4代替KCl;CK+NaHS:0.12 g/kg KCl,每周加10 mL 10 μmol/L NaHS;Al:2.5 g/kg AlCl3和0.12 g/kg KCl混在土中;Al+S:2.5 g/kg AlCl3、0.12 g/kg K2SO4代替KCl混在土中;Al+NaHS:2.5 g/kg AlCl3、0.12 g/kg KCl混在土中,每周加10 mL 10 μmol/L NaHS。下同。
Fig. 1. Changes of H2S content in rice under different treatments Different letters in the figure mean significant difference in the results by the analysis of variance (P<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage; HS, Full heading stage; MS, Mature stage. CK, 0.12 g/kg KCl; CK+S, KCl was replaced by 0.12 g/kg K2SO4;CK+NaHS, 0.12 g/kg KCl; 10 mL 10 μmol/L NaHS per week, Al, 2.5 g/kg AlCl3 was mixed with soil; 0.12 g/kg KCl, Al+S: 2.5 g/kg AlCl3 was mixed with soil; KCl was replaced by 0.12 g/kg K2SO4; Al+NaHS, 2.5 g/kg AlCl3 + 0.12 g/kg KCl(mixed with soil); 10 mL 10 μmol/L NaHS was applied per week. The same below.
处理 Treatment | 根总长 Total root length(cm) | 根平均直径 Average root diameter(mm) | 根总面积 Total root area(cm²) | 根总体积 Total volume of roots(cm3) | 根尖计数 Total root tip counts |
---|---|---|---|---|---|
CK | 5326.61±67.90 b | 1.48±0.16 a | 1779.40±80.11 b | 49.37±7.015 b | 33449±412 ab |
CK+S | 5363.48±7.31 b | 1.37±0.16 ab | 1775.80±81.29 b | 53.21±3.95 b | 29847±603 b |
CK+NaHS | 5373.04±79.76 b | 1.30±0.04 abc | 1526.82±93.32 c | 43.98±3.56 b | 30219±175 b |
Al | 3356.06±105.96 d | 1.06±0.03 c | 1052.38±7.75 d | 27.03±7.10 c | 20640±448 c |
Al+S | 6107.49±21.07 a | 1.17±0.15 bc | 2480.62±165.94 a | 71.50±7.90 a | 35210±4190 a |
Al+NaHS | 3559.97±139.96 c | 1.43±0.06 ab | 1779.40±80.11 b | 45.27±3.82 b | 23117±881 c |
表2 外源施加硫肥对水稻根系生长的影响
Table 2. Effects of S fertilizer application on rice root growth
处理 Treatment | 根总长 Total root length(cm) | 根平均直径 Average root diameter(mm) | 根总面积 Total root area(cm²) | 根总体积 Total volume of roots(cm3) | 根尖计数 Total root tip counts |
---|---|---|---|---|---|
CK | 5326.61±67.90 b | 1.48±0.16 a | 1779.40±80.11 b | 49.37±7.015 b | 33449±412 ab |
CK+S | 5363.48±7.31 b | 1.37±0.16 ab | 1775.80±81.29 b | 53.21±3.95 b | 29847±603 b |
CK+NaHS | 5373.04±79.76 b | 1.30±0.04 abc | 1526.82±93.32 c | 43.98±3.56 b | 30219±175 b |
Al | 3356.06±105.96 d | 1.06±0.03 c | 1052.38±7.75 d | 27.03±7.10 c | 20640±448 c |
Al+S | 6107.49±21.07 a | 1.17±0.15 bc | 2480.62±165.94 a | 71.50±7.90 a | 35210±4190 a |
Al+NaHS | 3559.97±139.96 c | 1.43±0.06 ab | 1779.40±80.11 b | 45.27±3.82 b | 23117±881 c |
处理 Treatment | 净光合速率 Pn(µmol·m−2s−1) | 胞间CO2浓度 Ci(mmol·mol−1) | 蒸腾速率 Tr(μmol·m−2s−1) | 气孔导度 Gs(μmol·m−2s−1) |
---|---|---|---|---|
CK | 13.51±0.30 a | 325.08±3.55 ab | 9.80±0.42 a | 384.64±21.84 a |
CK+S | 13.19±0.87 a | 315.18±5.36 bc | 9.21±0.70 a | 361.63±14.37 b |
CK+NaHS | 12.98±1.00 a | 313.5±0.02 bc | 9.50±0.53 a | 355.15±6.50 b |
Al | 6.66±0.13 d | 284.75±7.54 c | 3.64±0.31 d | 223.69±2.11 d |
Al+S | 8.44±0.05 c | 304.69±3.24 b | 4.94±0.63 c | 251.93±10.34 c |
Al+NaHS | 12.51±0.06 a | 334.13±6.60 a | 6.73±1.06 b | 356.34±3.11 b |
表3 施加硫肥对水稻光合指标的影响
Table 3. Effects of S fertilizer application on photosynthetic indexes of rice
处理 Treatment | 净光合速率 Pn(µmol·m−2s−1) | 胞间CO2浓度 Ci(mmol·mol−1) | 蒸腾速率 Tr(μmol·m−2s−1) | 气孔导度 Gs(μmol·m−2s−1) |
---|---|---|---|---|
CK | 13.51±0.30 a | 325.08±3.55 ab | 9.80±0.42 a | 384.64±21.84 a |
CK+S | 13.19±0.87 a | 315.18±5.36 bc | 9.21±0.70 a | 361.63±14.37 b |
CK+NaHS | 12.98±1.00 a | 313.5±0.02 bc | 9.50±0.53 a | 355.15±6.50 b |
Al | 6.66±0.13 d | 284.75±7.54 c | 3.64±0.31 d | 223.69±2.11 d |
Al+S | 8.44±0.05 c | 304.69±3.24 b | 4.94±0.63 c | 251.93±10.34 c |
Al+NaHS | 12.51±0.06 a | 334.13±6.60 a | 6.73±1.06 b | 356.34±3.11 b |
图2 外源施加硫肥对水稻体内光合色素含量的影响图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。
Fig. 2. Effects of S fertilizer application on photosynthetic pigment contents in rice Different letters in the figure mean significant difference in the results by the analysis of variance (P<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage.
图3 外源施加硫肥对水稻不同生育期根、茎和叶中Al含量的影响图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。
Fig. 3. Effects of S fertilizer application on Al contents in roots, stems and leaves of rice at different growth stages Different letters in the figure mean significant difference in the results by the analysis of variance (P<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage; HS, Full heading stage; MS, Mature stage.
图4 外源施加硫肥对根际土中不同Al形态和pH的影响图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。TS:分蘖期,HS:齐穗期,MS:成熟期。
Fig. 4. Effects of S application on the changes of Al form and pH in rhizosphere soil Different letters in the figure mean significant difference in the results by the analysis of variance (p<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage, HS, Full heading stage, MS, Mature stage. Alw,Water soluble aluminum; Alo, Organic combined aluminum; Alh, Humic acid aluminum; Aloh,Aluminum of hydrous oxide and hydroxide; Alino, Inorganic adsorption aluminum; Alex, Exchangeable aluminum.
图5 外源施加硫肥对不同生育期水稻根和叶中O2−·、MDA和H2O2含量的影响图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。
Fig. 5. Effects of S fertilizer application on contents of O2−·, MDA and H2O2 in roots and leaves of rice at different growth stages Different letters in the figure indicate significant difference in the results by the analysis of variance (P<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage; HS, Full heading stage; MS, Mature stage.
图6 外源施加硫肥对不同生育期水稻根和叶中抗氧化酶活性的影响图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。
Fig. 6. Effects of S fertilizer on the activities of antioxidant enzymes in root and leaf of rice at different growth stages Different letters in the figure indicate significant difference in the results by the analysis of variance (P<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage, HS, Full heading stage, MS, Mature stage.
图7 外源施加硫肥对不同生育期水稻体内OsALS1、OsSTAR1和OsSTAR2相对表达量的影响图中不同的字母表示方差分析下结果差异显著(P<0.05),数值为平均值±标准偏差(n=3)。
Fig. 7. Effects of exogenous application of S fertilizer on the relative expression levels of OsALS1, OsSTAR1 and OsSTAR2 in rice at different growth stages Different letters indicate significant difference by the analysis of variance (P<0.05), and the values are mean ± standard deviation (n=3). TS, Tillering stage; HS, Full heading stage; MS, Mature stage.
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