中国水稻科学 ›› 2016, Vol. 30 ›› Issue (6): 567-576.DOI: 10.16819/j.1001-7216.2016.6017
• • 下一篇
胡训霞1, 史春阳1, 丁艳1, 张萍1, 葛永胜1, 刘玉金2,*(), 王泽港1, 葛才林1,*()
收稿日期:
2016-02-02
修回日期:
2016-04-28
出版日期:
2016-11-10
发布日期:
2016-11-10
通讯作者:
刘玉金,葛才林
基金资助:
Xun-xia HU1, Chun-yang SHI1, Yan DING1, Ping ZHANG1, Yong-sheng GE1, Yu-jin LIU2,*(), Ze-gang WANG1, Cai-lin GE1,*()
Received:
2016-02-02
Revised:
2016-04-28
Online:
2016-11-10
Published:
2016-11-10
Contact:
Yu-jin LIU, Cai-lin GE
摘要:
研究水稻耐低磷机制,对高效利用土壤磷的水稻品种改良具有重要意义。应用基因芯片和qRT-PCR技术,分析了耐低磷水稻品种仪2434和低磷敏感品种通粳981根系中磷高效吸收和利用相关基因表达对低磷胁迫的应答。水稻叶片磷含量的测定结果表明,低磷胁迫下仪2434的叶片磷含量较对照降幅小,这表明仪2434较通粳981具有更强的磷素吸收利用能力。基因芯片检测结果表明,在低磷胁迫下的仪2434根系中,PHR1、osa-miR399s和SPXs基因的表达诱导、激活磷饥饿信号途径;APA、PAPs、MPE、PA、PEPC和VDAC1、C4-DT/MAT 等基因的表达诱导,增强有机磷水解酶和有机酸的合成和分泌,促进介质中难溶性磷的活化;OsPT2、OsPT6基因的表达诱导,促进仪2434根系对磷的高效吸收。qRT-PCR检测结果表明,仪2434根系中与磷饥饿信号转导、磷活化、磷高效吸收相关的8个代表性基因(PHR1、SPX、PAP、APA、PEPC、MFS、OsPT2、OsPT6)的表达水平均随低磷处理时间的延长呈逐步增高的趋势;经低磷处理后,所检测的8个基因在仪2434根系中的转录水平均显著高于其在通粳981根系中的转录水平,PHR1、APA、OsPT2在通粳981根系中的表达诱导作用不明显,且仪2434根系组织和根系分泌的酸性磷酸酶活性较通粳981增强更显著,这可能是仪2434较通粳981对低磷胁迫有较高耐性的主要机制之一。
中图分类号:
胡训霞, 史春阳, 丁艳, 张萍, 葛永胜, 刘玉金, 王泽港, 葛才林. 水稻根系中磷高效吸收和利用相关基因表达对低磷胁迫的应答[J]. 中国水稻科学, 2016, 30(6): 567-576.
Xun-xia HU, Chun-yang SHI, Yan DING, Ping ZHANG, Yong-sheng GE, Yu-jin LIU, Ze-gang WANG, Cai-lin GE. Response of Gene Expression Related to Efficient Phosphorus Absorption and Utilization to Low-P Stress in Rice Roots[J]. Chinese Journal OF Rice Science, 2016, 30(6): 567-576.
Gene ID 基因ID | 功能注释 Function annotation | 上游引物 Forward primer | 下游引物 Reverse primer |
---|---|---|---|
Os01g0239000 | PHR1 | 5'-CGCAAGGTGAAGGTGGACT-3' | 5'-CGATGTTGTGGCGAGTGAG-3' |
Os07g0614700 | SPX | 5'-CCCATCCAATGACCACC-3' | 5'-TTGAAAGCCAAAACACG-3' |
Os10g0116800 | PAP | 5'-ATCACTATGACTGGAGGGG-3' | 5'-TGTTTCTGCTGCTGATGTG-3' |
Os05g0192100 | APA | 5'-AGTAGCACAAAGCAGCAATA-3' | 5'-CGTTCAGCATCTCCGTC-3' |
Os01g0758300 | PEPC | 5'-TCCAAGCCGCCTTTAGAA-3' | 5'-ATCACGGTCTCCACCCATC-3' |
Os06g0324800 | MFS | 5'-CCCTACGATGGATACTGGC-3' | 5'-AGGATGAAGGTGGTGGTGTT-3' |
Os03g0150800 | QsPT2 | 5'-AGCAAGGTCGGGTGGAT-3' | 5'-GAAGGTGAGTGCGTAGAGC-3' |
Os08g0564000 | OsPT6 | 5'-GCCTGCTCTTCACCTTCC-3' | 5'-CCGACGACAACGACAAAA-3' |
表1 用于实时荧光定量PCR的特异性引物序列
Table 1 The primer sequences used for qRT-PCR verification.
Gene ID 基因ID | 功能注释 Function annotation | 上游引物 Forward primer | 下游引物 Reverse primer |
---|---|---|---|
Os01g0239000 | PHR1 | 5'-CGCAAGGTGAAGGTGGACT-3' | 5'-CGATGTTGTGGCGAGTGAG-3' |
Os07g0614700 | SPX | 5'-CCCATCCAATGACCACC-3' | 5'-TTGAAAGCCAAAACACG-3' |
Os10g0116800 | PAP | 5'-ATCACTATGACTGGAGGGG-3' | 5'-TGTTTCTGCTGCTGATGTG-3' |
Os05g0192100 | APA | 5'-AGTAGCACAAAGCAGCAATA-3' | 5'-CGTTCAGCATCTCCGTC-3' |
Os01g0758300 | PEPC | 5'-TCCAAGCCGCCTTTAGAA-3' | 5'-ATCACGGTCTCCACCCATC-3' |
Os06g0324800 | MFS | 5'-CCCTACGATGGATACTGGC-3' | 5'-AGGATGAAGGTGGTGGTGTT-3' |
Os03g0150800 | QsPT2 | 5'-AGCAAGGTCGGGTGGAT-3' | 5'-GAAGGTGAGTGCGTAGAGC-3' |
Os08g0564000 | OsPT6 | 5'-GCCTGCTCTTCACCTTCC-3' | 5'-CCGACGACAACGACAAAA-3' |
功能注释 Function annotation | 基因ID Gene ID | Log2倍数 Log2FC | P值 P-value |
---|---|---|---|
miR399 | osa-miR399d | 3.398±0.307 | 1.133×10-4 |
osa-miR399i | 2.572±0.173 | 8.032×10-5 | |
osa-miR399j | 3.184±0.115 | 1.027×10-6 | |
磷饥饿调控蛋白 Similar to phosphate starvation regulator protein (PHR1) | Os01g0239000 | 1.953±0.799 | 0.048 |
磷饥饿调控蛋白 Similar to phosphate starvation response regulator-like protein | Os02g0325600 | 2.144±0.080 | 4.665×10-4 |
SPX结构域 Oryza sativa SYG/PHO8/XPR1 (SPX) domain gene | Os06g0603600 | 1.412±0.271 | 9.934×10-4 |
SPX结构域 SPX N-terminal domain containing protein | Os07g0614700 | 4.475±0.241 | 2.628×10-4 |
Os10g0392600 | 2.218±0.263 | 4.952×10-5 | |
Os02g0202200 | 2.374±0.204 | 1.624×10-4 | |
磷酸盐1 Similar to PHO1-like protein | Os06g0493600 | 3.119±0.047 | 2.594×10-4 |
高亲和力磷载体2 High affinity phosphate transporter 2 (OsPht2) | Os03g0150800 | 1.395±0.426 | 1.964×10-3 |
磷载体6 Oryza sativa phosphate transporter 6 (OsPht6) | Os08g0564000 | 6.488±0.525 | 5.095×10-5 |
磷载体6 Similar to phosphate transporter 6(OsPht6-like) | Os04g0186400 | 1.297±0.119 | 0.040 |
紫色酸性磷酸酶 Purple acid phosphatase (PAP) | Os12g0576600 | 4.514±0.490 | 7.743×10-5 |
紫色酸性磷酸酶 Similar to purple acid phosphatase (PAP) | Os12g0637100 | 2.559±0.342 | 6.757×10-3 |
Os10g0116800 | 5.107±0.219 | 6.184×10-5 | |
Os11g0151700 | 3.024±0.376 | 3.155×10-3 | |
Os01g0776600 | 2.573±0.077 | 9.358×10-5 | |
紫色酸性磷酸酶 Purple acid phosphatase-like (PAP-like) | Os01g0941800 | 5.442±0.449 | 1.612×10-5 |
金属磷酸酯酶 Metallophosphoesterase domain protein (MPE) | Os07g0106000 | 3.055±0.603 | 2.646×10-4 |
植酸酶 Similar to Phytase (PA) | Os08g0280100 | 3.372±0.025 | 1.659×10-5 |
Os03g0848200 | 1.926±0.124 | 1.610×10-5 | |
酸性磷酸酶 Acid phosphatase (Class B) (APA) | Os05g0192100 | 1.353±0.075 | 1.420×10-3 |
磷酸烯醇式丙酮酸羧化酶 Similar toPhosphoenolpyruvate carboxylase (PEPC) | |||
Os01g0758300 | 1.791±0.860 | 0.017 | |
Os01g0110700 | 2.147±0.677 | 2.508×10-3 | |
C4-二羧酸转运/苹果酸转运蛋白 C4-dicarboxylate transporter/malic acid transport protein (C4-DT/MAT) | Os01g0226600 | 2.537±1.118 | 7.562×10-3 |
电压依赖阴离子通道1 Voltage-dependent anion channe1 (VDAC1) | Os01g0588200 | 1.276±0.240 | 4.216×10-4 |
协助转运超家族Major facilitator superfamily protein (MFS) | Os06g0324800 | 4.217±1.267 | 1.278×10-3 |
Os08g0409900 | 4.113±0.415 | 2.175×10-5 | |
Os08g0156600 | 3.465±0.131 | 4.637×10-5 |
表2 低磷胁迫对仪2434根系中与磷饥饿信号、磷活化和高效吸收相关基因表达的影响
Table 2 Effect of low-P stress on the expression of genes related to phosphorus starvation signals, phosphorus activation and high efficiency absorption.
功能注释 Function annotation | 基因ID Gene ID | Log2倍数 Log2FC | P值 P-value |
---|---|---|---|
miR399 | osa-miR399d | 3.398±0.307 | 1.133×10-4 |
osa-miR399i | 2.572±0.173 | 8.032×10-5 | |
osa-miR399j | 3.184±0.115 | 1.027×10-6 | |
磷饥饿调控蛋白 Similar to phosphate starvation regulator protein (PHR1) | Os01g0239000 | 1.953±0.799 | 0.048 |
磷饥饿调控蛋白 Similar to phosphate starvation response regulator-like protein | Os02g0325600 | 2.144±0.080 | 4.665×10-4 |
SPX结构域 Oryza sativa SYG/PHO8/XPR1 (SPX) domain gene | Os06g0603600 | 1.412±0.271 | 9.934×10-4 |
SPX结构域 SPX N-terminal domain containing protein | Os07g0614700 | 4.475±0.241 | 2.628×10-4 |
Os10g0392600 | 2.218±0.263 | 4.952×10-5 | |
Os02g0202200 | 2.374±0.204 | 1.624×10-4 | |
磷酸盐1 Similar to PHO1-like protein | Os06g0493600 | 3.119±0.047 | 2.594×10-4 |
高亲和力磷载体2 High affinity phosphate transporter 2 (OsPht2) | Os03g0150800 | 1.395±0.426 | 1.964×10-3 |
磷载体6 Oryza sativa phosphate transporter 6 (OsPht6) | Os08g0564000 | 6.488±0.525 | 5.095×10-5 |
磷载体6 Similar to phosphate transporter 6(OsPht6-like) | Os04g0186400 | 1.297±0.119 | 0.040 |
紫色酸性磷酸酶 Purple acid phosphatase (PAP) | Os12g0576600 | 4.514±0.490 | 7.743×10-5 |
紫色酸性磷酸酶 Similar to purple acid phosphatase (PAP) | Os12g0637100 | 2.559±0.342 | 6.757×10-3 |
Os10g0116800 | 5.107±0.219 | 6.184×10-5 | |
Os11g0151700 | 3.024±0.376 | 3.155×10-3 | |
Os01g0776600 | 2.573±0.077 | 9.358×10-5 | |
紫色酸性磷酸酶 Purple acid phosphatase-like (PAP-like) | Os01g0941800 | 5.442±0.449 | 1.612×10-5 |
金属磷酸酯酶 Metallophosphoesterase domain protein (MPE) | Os07g0106000 | 3.055±0.603 | 2.646×10-4 |
植酸酶 Similar to Phytase (PA) | Os08g0280100 | 3.372±0.025 | 1.659×10-5 |
Os03g0848200 | 1.926±0.124 | 1.610×10-5 | |
酸性磷酸酶 Acid phosphatase (Class B) (APA) | Os05g0192100 | 1.353±0.075 | 1.420×10-3 |
磷酸烯醇式丙酮酸羧化酶 Similar toPhosphoenolpyruvate carboxylase (PEPC) | |||
Os01g0758300 | 1.791±0.860 | 0.017 | |
Os01g0110700 | 2.147±0.677 | 2.508×10-3 | |
C4-二羧酸转运/苹果酸转运蛋白 C4-dicarboxylate transporter/malic acid transport protein (C4-DT/MAT) | Os01g0226600 | 2.537±1.118 | 7.562×10-3 |
电压依赖阴离子通道1 Voltage-dependent anion channe1 (VDAC1) | Os01g0588200 | 1.276±0.240 | 4.216×10-4 |
协助转运超家族Major facilitator superfamily protein (MFS) | Os06g0324800 | 4.217±1.267 | 1.278×10-3 |
Os08g0409900 | 4.113±0.415 | 2.175×10-5 | |
Os08g0156600 | 3.465±0.131 | 4.637×10-5 |
图2 仪2434(YI2434)和通粳981(TJ981)根系中与磷饥饿信号、磷活化和高效吸收相关基因转录水平随时间变化的qRT-PCR验证结果 A, B, C, D, E, F, G, H 分别表示PHR1 (Os01g0239000)、 SPXs (Os07g0614700)、 PAPs (Os10g0116800)、 APA (Os05g0192100)、 PEPC (Os01g0758300)、 MFSs (Os06g0324800)、 OsPT2 (Os03g0150800)、 OsPT6 (Os08g0564000)基因。*、**分别表示在P<0.05和P<0.01水平上差异显著。
Fig. 2. Transcription level of phosphorus starvation signals, phosphorus activation and high efficiency absorption related gene changes with low-P treated time in YI2434 and TJ981 roots by qRT-PCR. A, B, C, D, E, F, G, H represent the genes of PHR1 (Os01g0239000), SPXs (Os07g0614700), PAPs (Os10g0116800), APA (Os05g0192100), PEPC (Os01g0758300), MFSs (Os06g0324800), OsPT2 (Os03g0150800), OsPT6 (Os08g0564000), respectively. *, **represent significant difference at P < 0.05 and P < 0.01, respectively.
图3 低磷胁迫15 d仪2434和通粳981根系酸性磷酸酶活性。 *、**分别表示在P<0.05和P<0.01水平上差异显著。
Fig. 3. The acid phosphatase activity in YI2434 and TJ981 at 15 days after low-P treatment. *, **show significant difference at P < 0.05 and P < 0.01, respectively.
图4 低磷胁迫15 d仪2434和通粳981根系分泌酸性磷酸酶活性 *、**分别表示在P<0.05和P<0.01水平上差异显著。
Fig. 4. Acid phosphatase activity secreted by roots in YI2434 and TJ981 at 15 days after low-P treatment. *, **shows significant difference at P < 0.05 and P < 0.01,respectively.
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