硫化氢提高水稻磷吸收转运的生理和分子机制

doi:10.16819/j.1001-7216.2019.9055

摘要/Abstract

摘要：

目的低磷胁迫是限制水稻产量的主要因素之一。水稻淹水条件下产生H₂S,然而,H₂S作为信号分子是否参与调节水稻响应缺磷胁迫还未可知。方法在正常磷和低磷条件下测定水稻H₂S含量,揭示H₂S在水稻响应缺磷胁迫中的作用。用2 μmol/L H₂S前体物质NaHS预处理水稻1 d,然后在加磷和低磷条件下培养6 d,测定水稻体内总磷含量、酸性磷酸酶活性、抗氧化酶活性、木质部汁液磷含量、磷转运子基因表达以及根系构型变化,从而探究H₂S参与调节水稻响应缺磷胁迫的生理和分子机制。结论低磷胁迫下,水稻根系和地上部H₂S含量显著增加。NaHS预处理水稻显著增加低磷条件下水稻体内有效磷和总磷含量,提高根系酸性磷酸酶活性,提高抗氧化酶活性、木质部汁液磷含量和磷转运子基因表达水平,同时还改变水稻根系构型,增加总根长、总根表面积、总根体积和总根尖数,从而促进低磷条件下水稻对外界磷的吸收和转运,最终缓解缺磷胁迫。

关键词: 水稻, 低磷胁迫, 硫化氢, 磷转运子, 根系构型

Abstract:

【Objective】 Low phosphorus (P) stress is one of the main factors limiting rice yield. Hydrogen sulfide (H₂S) is produced in rice under flooding conditions. However, whether H₂S is involved in the regulation of rice response to phosphorus deficiency stress as a signal molecule is still unclear. 【Method】 The content of H₂S in rice was determined under normal and low P conditions to illuminate the role that H₂S plays in regulating rice response to P deficiency stress. Rice was pretreated with 2 μmol/L H₂S precursor NaHS for 1 day, then cultured for another 6 days under P sufficient and deficient conditions. The total P contents, acid phosphatase activity, antioxidant enzyme activity, xylem P concentration, P transporter genes’ expression and rice root architecture were determined to explore the physiological and molecular mechanism of H₂S in regulating rice response to phosphorus deficiency stress. 【Conclusion】 Low P stress significantly increased the content of H₂S in rice roots and shoots. NaHS pretreatment significantly increased total P contents, root acid phosphatase activity, root antioxidant enzymes activity, xylem P concentration and stimulated the expression of P transporter genes. Meanwhile, NaHS pretreatment modified rice root architecture, including increased total root length, total root surface area, total root volume and total root tip numbers. Therefore, our present study demonstrated that H₂S improved P absorption and translocation in rice under P deficiency conditions and finally alleviate P deficiency stress in rice.

Key words: rice, P deficiency stress, H₂S, P transporter, root architecture

中图分类号:

朱春权, 曹小闯, 朱练峰, 白志刚, 黄洁, 梁清铎, 金千瑜, 张均华. 硫化氢提高水稻磷吸收转运的生理和分子机制[J]. 中国水稻科学, 2019, 33(6): 532-540.

Chunquan ZHU, Xiaochuang CAO, Lianfeng ZHU, Zhigang BAI, Jie HUANG, Qingduo LIANG, Qianyu JIN, Junhua ZHANG. Physiological and Molecular Mechanisms of Hydrogen Sulfide Enhancing Phosphorus Absorption and Transportation in Rice[J]. Chinese Journal OF Rice Science, 2019, 33(6): 532-540.

图/表 9

表1 本研究中使用的引物序列

Table 1 Sequence of primers used in present study.

基因 Gene	正向引物（5′-3′） Forward (5′-3′)	反向引物（5′-3′） Reverse (5′-3′)
OsPT1	AGCGTTCGGGTTCCTGTA	CGTTCTTGATGCCGATCC
OsPT2	GACGAGACCGCCCAAGAAG	TTTTCAGTCACTCACGTCGAGAC
OsPT3	GTGCTCATGGTGGTGTGCT	GAGCCAGAACCGGAAGAAG
OsPT4	GGAGAAGGCTGACGAGGTC	CCCATGGCGTCTCAAAAA
OsPT5	GGCGAGAACGAAATGGAG	GACGGTCTGCCTGTAGGAGT
OsPT6	TATAACTGATCGATCGAGACCAGAG	TGGATAGCCAGGCCAGTTATATATC
OsPT7	GCTTCCTCCTCACCTTCCTT	TTCTCCCGTGACATCTCCTC
OsPT8	AGAAGGCAAAAGAAATGTGTGTTAAAT	AAAATGTATTCGTGCCAAATTGCT
OsPT9	CATAGGCTTGTCATCCTTTGG	CACTGTAAATAAATCCGCGTTTC
OsPT10	GAGCTCGCACCTCAGCAT	GAGTTCACTCACACGGAGACC
OsPT12	AAATCGAGGTGGAGGAGGAG	CGAGAAGAGGCCGTAGTCC
OsHistone	GGTCAACTTGTTGATTCCCCTCT	AACCGCAAAATCCAAAGAACG

图 1 不同缺磷时间段水稻根系和地上部的硫化氢含量数据用均值±标准差(n = 4)表示。数据用均值±标准差(n = 4)表示。不同的小写字母代表处理间差异在0.05水平上显著。缺磷处理的磷浓度为18 mmol/L。

Fig. 1. Content of H2S in rice root and shoot under different phosphorus deficient durations. Data are means ± SD (n = 4). Different letters stand for significant difference at P < 0.05. The P concentration in the nutrient solution is 18 mmol/L under P deficient conditions.

图2 水稻根部和地上部的总磷含量数据用均值±标准差(n = 4)表示。不同的小写字母代表处理间差异在P < 0.05水平上显著。P–营养液中磷浓度为180 mmol/L, LP–18 mmol/L。

Fig. 2. Available P and total P contents in rice roots and shoots. Data are means ± SD (n = 4). Different letters above the columns mean significant difference at P < 0.05. P, P concentration of 180 mmol/L, LP, 18 mmol/L.

图3 水稻根部和地上部的酸性磷酸酶活性数据用均值±标准差(n = 4)表示。不同的小写字母代表处理间差异在P < 0.05水平上显著。P–180 mmol/L; LP–18 mmol/L。

Fig. 3. Activites of acid phosphatase in rice roots and shoots. Data are means ± SD (n = 4). Different letters above the columns are significantly different at P < 0.05. P, P concentration of 180 mmol/L; LP, 18 mmol/L.

图4 水稻木质部汁液磷含量数据用均值±标准差(n = 4)表示。不同的小写字母代表处理间差异在P < 0.05水平上显著。P–180 mmol/L, LP–18 mmol/L。

Fig. 4. Xylem P concentration in rice. Data are means ± SD (n = 4). Different letters above the bars mean significant difference at P < 0.05. P, 180 mmol/L, LP, 18 mmol/L.

图5 磷转运子基因相对表达量数据用均值±标准差(n = 4)表示。不同的小写字母代表处理间差异在P < 0.05水平上显著。P–180 mmol/L, LP–18 mmol/L。

Fig. 5. Relative expression level of P transporter genes. Data are means ± SD (n = 4). Different letters above the bars mean significant difference at P < 0.05. P, 180 mmol/L, LP, 18 mmol/L.

表2 水稻根系和地上抗氧化酶活性

Table 2 Activities of antioxidant enzymes in rice roots and shoots.

酶的名称 Enzyme name	部位 Position	处理 Treatment
酶的名称 Enzyme name	部位 Position	正常磷 P	磷+硫氢化钠 P+NaHS	低磷 LP	低磷+硫氢化钠 LP+NaHS
超氧化物歧化酶 Superoxide dismutase/(U·g^-1)	根部 Roots	3.39±0.08 d	3.89±0.39 c	6.21±0.88 b	10.74±1.24 a
超氧化物歧化酶 Superoxide dismutase/(U·g^-1)	地上部 Shoots	35.82±2.61 d	49.44±5.17 c	63.32±5.00 b	73.20±2.62 a
过氧化物酶 Peroxidase/(U·g^-1)	根部 Roots	837.56±35.96 d	1300.92±74.56 c	1852.22±160.51 b	2601.40±94.97 a
过氧化物酶 Peroxidase/(U·g^-1)	地上部 Shoots	1745.27±346.02 d	2649.98±178.02 c	5070.26±639.59 b	7627.11±688.69 a
抗坏血酸过氧化物酶 Ascorbate peroxidase/(μmol·min^-1 g^-1)	根部 Roots	0.17±0.04 c	0.47±0.10 b	0.45±0.08 b	0.62±0.07 a
抗坏血酸过氧化物酶 Ascorbate peroxidase/(μmol·min^-1 g^-1)	地上部 Shoots	0.26±0.06 d	0.45±0.04 c	0.65±0.09 b	0.92±0.22 a
过氧化氢酶 Catalase/(nmol·min^-1 g^-1)	根部 Roots	10.56±0.16 d	12.90±0.14 c	17.51±0.28 b	22.79±1.83 a
过氧化氢酶 Catalase/(nmol·min^-1 g^-1)	地上部 Shoots	7.08±2.44 d	23.23±3.88 c	36.30±4.57 b	63.21±16.26 a

表3 水稻根系发育相关指标

Table 3 Root development parameters in rice.

参数 Parameter	正常磷 P	磷+硫氢化钠 P+NaHS	低磷 LP	低磷+硫氢化钠 LP+NaHS
总根长Total length/cm	234.77±6.63 c	262.77±2.27 b	269.38±5.49 b	309.72±8.28 a
总表面积Total surface area/cm²	31.18±1.00 c	35.60±1.12 b	35.76±0.69 b	45.37±3.63 a
平均直径Average diagram/cm	0.42±0.01 a	0.43±0.01 a	0.43±0.01 a	0.43±0.01 a
根系总体积Root volume/cm³	0.35±0.01 c	0.39±0.01 b	0.39±0.00 b	0.45±0.03 a
根尖数Root tip number	448.0±15.0 d	392.0±20.0 c	549.0±5.3 b	639.7±23.2 a

图6 水稻根系表型

Fig. 6. Phenotype of rice roots.

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