一氧化氮处理下水稻幼苗叶片全基因组差异基因表达谱分析

发布时间：2018-12-15 23:58

【摘要】：一氧化氮(Nitric oxide,NO)是一种带自由基性质的气态激素,具有高度反应活性和膜透性。植物不仅对外源NO作出反应,而且能自身合成NO。近年来的研究发现,NO在植物生长、发育及胁迫响应中发挥重要作用。基于高通量DNA芯片技术的基因表达谱分析已被广泛应用于植物生长、发育和胁迫响应机制研究中,并取得了重大进展。然而,NO处理下水稻全基因组差异基因的表达谱分析未见报道。本研究采用水稻全基因组cDNA微阵列(Agilent rice oligo microarray,4×44k),对NO处理后1(T1)、6(T6)和12 h(T12)水稻幼苗叶片全基因组基因的表达谱进行了分析。结果表明,在任1个时间点有3倍或以上表达变化的基因有3852个,在3个时间点中至少两个时间点有3倍或以上表达变化的NO响应基因1195个。基因功能分析和代谢通路分析表明,NO响应基因主要参与了胁迫响应、氧化还原、次级代谢、生物合成等生理过程。实时荧光定量PCR验证结果与芯片杂交结果基本一致,印证了芯片杂交结果的有效性。该研究为阐明NO在植物生理过程中的作用机制提供了理论依据。NO信号分子与WRKY转录因子均参与植物抗逆、发育与代谢等许多生理过程。为了探讨NO信号是否参与WRKY蛋白介导的生理调节功能,本研究另对NO处理下水稻WRKY基因家族的表达谱作了系统分析。鉴定出在1个时间点有两倍或以上表达变化的差异表达的WRKY基因32个,主要分布在WRKY的I和II组,其中75%的IIa和45.6%的IId亚组成员为差异表达基因。鉴定出3个时间点至少在2个时间点有两倍或以上表达变化的NO响应的WRKY基因15个,均为早期(1 h)应答,且多数(64.2%)持续上调。基因功能预测分析表明,NO响应的WRKY基因主要参与生物学过程中的代谢过程、刺激响应和细胞过程,以及分子功能中的转录调节活性和结合。代谢通路分析表明,WRKY24参与植物与病原菌互作代谢通路。上述发现提示,NO信号可能参与了WRKY转录因子介导的生物学调控功能,并为下一步这些WRKY基因的功能分析奠定了基础。
[Abstract]:Nitric oxide (Nitric oxide,NO) is a free radical gaseous hormone with high reactivity and membrane permeability. Plants not only react to exogenous NO, but also synthesize NO. themselves. Recent studies have found that NO plays an important role in plant growth, development and stress response. Gene expression profiling based on high-throughput DNA microarray has been widely used in plant growth, development and stress response mechanisms, and has made great progress. However, NO treatment of rice genome differential gene expression profile analysis has not been reported. In this study, the whole genome cDNA microarray (Agilent rice oligo microarray,4 脳 44k was used to analyze the whole genome gene expression profiles of 1 (T1), 6 (T6) and 12 h (T12) leaves of rice seedlings treated with NO. The results showed that 3 852 genes had 3 times or more expression changes at any one time point, and 1195 NO response genes with 3 times or more expression changes at least 2 time points in 3 time points. Gene function analysis and metabolic pathway analysis showed that NO response genes were mainly involved in stress response, redox, secondary metabolism, biosynthesis and other physiological processes. The results of real-time fluorescence quantitative PCR were consistent with those of chip hybridization, which confirmed the validity of chip hybridization. This study provides a theoretical basis for elucidating the mechanism of NO in plant physiological processes. Both NO signaling molecules and WRKY transcription factors are involved in many physiological processes such as plant stress resistance, development and metabolism. In order to investigate whether NO signal is involved in the physiological regulation mediated by WRKY protein, the expression profiles of WRKY gene family in rice treated with NO were analyzed systematically in this study. 32 differentially expressed WRKY genes were identified at one time point, mainly distributed in WRKY I and II groups. 75% of IIa and 45.6% of IId subgroup members were differentially expressed genes. At least 2 time points, 15 WRKY genes were identified, all of which were early (1 h) responsive, and most (64.2%) were up-regulated. Gene function prediction analysis showed that the WRKY gene of NO response was mainly involved in metabolic process, stimulatory response and cellular process, as well as transcriptional regulatory activity and binding in molecular function. Metabolic pathway analysis showed that WRKY24 was involved in the interaction of plant and pathogen. These findings suggest that NO signaling may be involved in the biological regulation mediated by WRKY transcription factors and may lay a foundation for the functional analysis of these WRKY genes in the next step.
【学位授予单位】：湖南科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2

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