一氧化氮處理下水稻幼苗葉片全基因組差異基因表達(dá)譜分析
發(fā)布時(shí)間：2018-12-15 23:58
【摘要】：一氧化氮(Nitric oxide,NO)是一種帶自由基性質(zhì)的氣態(tài)激素,具有高度反應(yīng)活性和膜透性。植物不僅對(duì)外源NO作出反應(yīng),而且能自身合成NO。近年來的研究發(fā)現(xiàn),NO在植物生長(zhǎng)、發(fā)育及脅迫響應(yīng)中發(fā)揮重要作用。基于高通量DNA芯片技術(shù)的基因表達(dá)譜分析已被廣泛應(yīng)用于植物生長(zhǎng)、發(fā)育和脅迫響應(yīng)機(jī)制研究中,并取得了重大進(jìn)展。然而,NO處理下水稻全基因組差異基因的表達(dá)譜分析未見報(bào)道。本研究采用水稻全基因組cDNA微陣列(Agilent rice oligo microarray,4×44k),對(duì)NO處理后1(T1)、6(T6)和12 h(T12)水稻幼苗葉片全基因組基因的表達(dá)譜進(jìn)行了分析。結(jié)果表明,在任1個(gè)時(shí)間點(diǎn)有3倍或以上表達(dá)變化的基因有3852個(gè),在3個(gè)時(shí)間點(diǎn)中至少兩個(gè)時(shí)間點(diǎn)有3倍或以上表達(dá)變化的NO響應(yīng)基因1195個(gè)�；蚬δ芊治龊痛x通路分析表明,NO響應(yīng)基因主要參與了脅迫響應(yīng)、氧化還原、次級(jí)代謝、生物合成等生理過程。實(shí)時(shí)熒光定量PCR驗(yàn)證結(jié)果與芯片雜交結(jié)果基本一致,印證了芯片雜交結(jié)果的有效性。該研究為闡明NO在植物生理過程中的作用機(jī)制提供了理論依據(jù)。NO信號(hào)分子與WRKY轉(zhuǎn)錄因子均參與植物抗逆、發(fā)育與代謝等許多生理過程。為了探討NO信號(hào)是否參與WRKY蛋白介導(dǎo)的生理調(diào)節(jié)功能,本研究另對(duì)NO處理下水稻W(wǎng)RKY基因家族的表達(dá)譜作了系統(tǒng)分析。鑒定出在1個(gè)時(shí)間點(diǎn)有兩倍或以上表達(dá)變化的差異表達(dá)的WRKY基因32個(gè),主要分布在WRKY的I和II組,其中75%的IIa和45.6%的IId亞組成員為差異表達(dá)基因。鑒定出3個(gè)時(shí)間點(diǎn)至少在2個(gè)時(shí)間點(diǎn)有兩倍或以上表達(dá)變化的NO響應(yīng)的WRKY基因15個(gè),均為早期(1 h)應(yīng)答,且多數(shù)(64.2%)持續(xù)上調(diào)�；蚬δ茴A(yù)測(cè)分析表明,NO響應(yīng)的WRKY基因主要參與生物學(xué)過程中的代謝過程、刺激響應(yīng)和細(xì)胞過程,以及分子功能中的轉(zhuǎn)錄調(diào)節(jié)活性和結(jié)合。代謝通路分析表明,WRKY24參與植物與病原菌互作代謝通路。上述發(fā)現(xiàn)提示,NO信號(hào)可能參與了WRKY轉(zhuǎn)錄因子介導(dǎo)的生物學(xué)調(diào)控功能,并為下一步這些WRKY基因的功能分析奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：湖南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2
本文編號(hào)：2381503
[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.
【學(xué)位授予單位】：湖南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2
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