• ISSN 1008-505X
  • CN 11-3996/S

苗期水稻响应低磷胁迫的分子调控机制

Molecular regulatory mechanisms of rice response to low phosphorus stress at seedling stage

  • 摘要:
    目的 磷素是水稻生长必需的大量营养元素之一,磷营养供应不足将影响水稻产量和稻米品质。研究低磷条件下水稻根系的生理响应及其基因的差异表达,明确水稻根系差异表达基因对磷高效吸收和利用的生理调控机制,旨在为农业生产中磷高效吸收利用水稻品种的选种和育种提供理论依据。
    方法 采用水培试验,供试水稻材料包括磷敏感基因型通粳981 (TJ981)、耐低磷基因型郑旱6号(ZH6)和根系扩展型镇稻99 (ZD99)。分别设置营养液正常(CK)与低磷(LP,磷浓度为CK的1/20) 两个磷浓度。水稻生长15天后取样,采用转录组测序(RNA-seq)技术,结合水稻根内磷含量、根系分泌酸性磷酸酶(APase)活性及根际pH等指标变化情况,分析差异表达基因在水稻根系生理调控中的作用。
    结果 与CK处理相比,LP处理的通粳981、镇稻99和郑旱 6号根系上、下调差异表达基因数分别是447和1395个、352和1185个、475和1856个,且3个不同水稻品种共有237个差异表达基因,其中33个上调差异表达基因和204个下调差异表达基因。低磷条件下,通粳981、镇稻99和郑旱 6号水稻根中磷含量均显著低于对照(CK),分别为对照的41.31%、36.29% 和33.17%,高于两处理培养液中磷含量的差异,表明无机磷转运蛋白1-2基因的上调表达促进了根系对磷的吸收。低磷处理1、8和15天时,3个品种水稻根系分泌酸性磷酸酶(Apase)活性均高于对照,这一结果与紫色酸性磷酸酶基因的差异表达有关。
    结论 低磷诱导水稻根系基因的差异表达,水稻根系中转录下调基因数远多于转录上调基因数;低磷诱导水稻根系中无机磷转运蛋白OsPT1-2的上调表达,促进低磷条件下水稻根系对磷的吸收;紫色酸性磷酸酶和质膜H+-ATP酶基因的诱导表达,增加了水稻根系分泌酸性磷酸酶活性,降低了H+的运输和分泌,导致根际pH降低,更有利于磷的吸收。

     

    Abstract:
    Objectives Phosphorus (P) is one of the essential nutrients for rice growth, and insufficient phosphorus supply will affect rice yield and quality. We studied the physiological response and differential expression of genes in rice roots to low phosphorus conditions, aiming to serve the selection and breeding of rice varieties with efficient phosphorus absorption and utilization in agricultural production.
    Methods A phosphorus sensitive genotype Tongjing 981 (TJ981), a low phosphorus tolerant genotype Zhenghan 6 (ZH6), and a root extension type Zhendao 99 (ZD99) were selected as the research materials in a hydroponic experiment. Normal and low phosphorous nutrition solution was prepared as control (CK) and low P treatment (LP), the P concentration in LP solution was 5% of that in +P solution. At 1, 8, and 15 days of treatment, rice seedlings were sampled for measurement. The differentially expressed genes (DEGs) were analyzed using transcriptome sequencing (RNA seq) technology, protein mass spectrometry analysis, and real-time fluorescence quantitative PCR (RT-PCR) technology; the root phosphorus content and acid phosphatase (APase) activity, and rhizosphere pH were measured at the same time.
    Results LP treatment increased the number of DEGs on the roots of all the three rice varieties, the up- and down-regulated DEGs were 447 and 1395 for Tongjing 981, 352 and 1185 for Zhendao 99, and 475 and 1856 for Zhenghan 6, respectively. There were a total of 237 DEGs that identified in all the three varieties, of them, 33 were upregulated and 204 were downregulated. The P contents in the roots of rice variety Tongjing 981, Zhendao 99, and Zhenghan 6 under LP condition were 41.31%, 36.29%, and 33.17% of those under +P condition, the decrease exceeded the P concentration gap between the two culture solutions, indicating that the upregulation of the expression of the inorganic phosphorus transporter 1-2 gene promoted the absorption of phosphorus by roots. At the 1, 8, and 15 days of LP treatment, the activity of APase of three rice varieties were all higher than those of +P, which could be explained by the variation of the differential expression of the purple acid phosphatase gene.
    Conclusions The number of superimposed inhibited genes was significantly higher than that of induced genes, that is, the number of down-regulated genes was much more than that of up-regulated genes. By inducing the expression of inorganic phosphorus transporter OsPT1-2 in rice roots, the uptake of available phosphorus in low-phosphorus rice roots was promoted. By inducing the expression of purple acid phosphatase and plasma membrane H+-ATpase genes in rice roots, the activity of acid phosphatase secreted by rice roots increased, and H+ transport and secretion led to a decrease in rhizosphere pH, thus promoting the activation of organic and inorganic binding phosphorus and improving the efficient utilization of phosphorus.

     

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