本文選題：缺硫脅迫 + 水稻��； 參考：《河南師范大學(xué)》2017年碩士論文

【摘要】：水稻是全球最重要的谷類作物之一,同時(shí)也是重要飼料、纖維以及生物燃料來(lái)源。硫元素(S)作為植物必需營(yíng)養(yǎng)元素在植物生長(zhǎng)、發(fā)育、代謝過(guò)程以及產(chǎn)量中都起著重要的作用。硫最初主要以硫酸鹽的形式從環(huán)境中吸收,是蛋白質(zhì)、維生素、輔酶、谷胱甘肽或鐵氧還蛋白等多種物質(zhì)的組成成分,對(duì)還原小分子化合物以及重金屬、外源性物質(zhì)的去毒等起著至關(guān)重要的生化作用。近年來(lái)由于大量氮肥的施用以及含硫廢氣的排放限制,土壤缺硫即將成為限制農(nóng)業(yè)生產(chǎn)的一個(gè)因素。有關(guān)硫的吸收、轉(zhuǎn)運(yùn)和同化研究得較為清楚。擬南芥中缺硫的應(yīng)答機(jī)理研究得也較為清楚,而水稻對(duì)缺硫的應(yīng)答研究報(bào)道較少。因此,本研究將兩周齡的水稻幼苗進(jìn)行缺硫脅迫處理,觀察了缺硫脅迫對(duì)水稻植株生長(zhǎng)和生化水平的影響;并對(duì)正常生長(zhǎng)與缺硫脅迫一周的水稻葉子和根中轉(zhuǎn)錄組進(jìn)行對(duì)比分析,找到了水稻中缺硫脅迫應(yīng)答基因,并對(duì)其進(jìn)行差異表達(dá)分析,從而達(dá)到對(duì)水稻中缺硫脅迫相關(guān)基因表達(dá)調(diào)控機(jī)制探究的目的。具體研究結(jié)果如下:(1)生長(zhǎng)和生化水平的研究表明,缺硫脅迫兩周左右的水稻幼苗POD、CAT、SOD酶活力以及葉綠素含量開始出現(xiàn)顯著下降,還原型谷胱甘肽含量在第三周開始出現(xiàn)下降。約在缺硫處理一個(gè)月左右植株生長(zhǎng)表型表現(xiàn)出根長(zhǎng)明顯伸長(zhǎng)葉片較黃等表型。與擬南芥、油菜、楊樹等植物相比,水稻對(duì)缺硫脅迫不太敏感。(2)通過(guò)對(duì)正常及缺硫處理一周的水稻幼苗進(jìn)行轉(zhuǎn)錄組分析,葉中對(duì)照組檢測(cè)到20705條基因,處理組中檢測(cè)到31200個(gè)基因,根中對(duì)照組檢測(cè)到29964個(gè)基因,處理組檢測(cè)到31050條基因,說(shuō)明在缺硫脅迫下水稻幼苗中有眾多基因被誘導(dǎo)表達(dá)。應(yīng)用NOISeq技術(shù)對(duì)差異表達(dá)基因進(jìn)行檢測(cè),葉中篩選出866個(gè)差異表達(dá)基因,11個(gè)基因下調(diào)、855個(gè)基因上調(diào),根中共有289個(gè)差異表達(dá)基因,28個(gè)基因下調(diào)、261個(gè)基因上調(diào),且根與葉子中共同的差異表達(dá)基因很少。并對(duì)13個(gè)差異表達(dá)基因運(yùn)用實(shí)時(shí)熒光定量PCR進(jìn)行驗(yàn)證,得到了與RNA-seq相似的結(jié)果,說(shuō)明本次高通量測(cè)序的可靠性。(3)對(duì)差異表達(dá)基因進(jìn)行KEGG、Pathway顯著性富集分析表明,在缺硫脅迫下大量與氧化脅迫應(yīng)答相關(guān)的基因發(fā)生了差異表達(dá),葉差異表達(dá)基因主要與核酸代謝、信號(hào)轉(zhuǎn)導(dǎo)等多種代謝通路有關(guān);根中差異表達(dá)基因主要與能量代謝、信號(hào)轉(zhuǎn)導(dǎo)等多種代謝通路有關(guān)。(4)缺硫脅迫下負(fù)責(zé)硫運(yùn)輸?shù)牧蜣D(zhuǎn)運(yùn)蛋白的調(diào)控與擬南芥相似,表明水稻采用與擬南芥相似的機(jī)制來(lái)調(diào)控硫酸鹽的重新分布;與擬南芥一樣,水稻中硫同化途徑中的ATPS、APR、MSA1、SDI1、SAM以及miR395靶基因也受到不同程度的調(diào)控。但是,在擬南芥中缺硫脅迫可以導(dǎo)致miR395的靶基因ATPS1、ATPS3、ATPS4的表達(dá)水平下降,但水稻中ATPS的表達(dá)被誘導(dǎo),這很可能是單子葉和雙子葉植物調(diào)控機(jī)制的差異性。同時(shí)預(yù)測(cè)了Os09g27660很可能是miR395的靶基因。(5)對(duì)根和葉中差異表達(dá)轉(zhuǎn)錄因子分析表明,葉中有21個(gè)差異表達(dá)轉(zhuǎn)錄因子,根中有4個(gè)差異表達(dá)轉(zhuǎn)錄因子,葉中差異表達(dá)轉(zhuǎn)錄因子數(shù)目明顯高于根,這些轉(zhuǎn)錄因子對(duì)于調(diào)控缺硫應(yīng)答基因的表達(dá)及植物激素代謝具有重要意義。同時(shí)發(fā)現(xiàn)多個(gè)miRNA靶基因的表達(dá)發(fā)生明顯變化,說(shuō)明除miR395外其它miRNA很可能也參與了缺硫脅迫的應(yīng)答。本研究表明缺硫脅迫能促進(jìn)水稻根的生長(zhǎng);同時(shí)水稻幼苗的硫轉(zhuǎn)運(yùn)機(jī)制和同化途徑發(fā)生了相應(yīng)的變化來(lái)應(yīng)答硫匱乏的生理狀態(tài);缺硫脅迫同時(shí)誘導(dǎo)了氧化應(yīng)激反應(yīng),大量的氧化脅迫相關(guān)的應(yīng)答基因被調(diào)控;并得到一系列與水稻幼苗缺硫脅迫應(yīng)答相關(guān)的差異表達(dá)基因。該研究為水稻缺硫脅迫應(yīng)答調(diào)控網(wǎng)絡(luò)的進(jìn)一步研究奠定了基礎(chǔ),也將為提高水稻抗脅迫能力和產(chǎn)量的研究提供理論依據(jù),同時(shí)對(duì)于如何操縱硫代謝,幫助提高水稻等農(nóng)作物產(chǎn)量具有重要意義。
[Abstract]:Rice is one of the most important cereals in the world, and is also an important source of feed, fiber and biofuel. Sulfur element (S) plays an important role in plant growth, development, metabolism and production as essential nutrients in plants. Sulphur is originally absorbed from the environment in the form of sulphate, which is a protein, vitamin, The components of a variety of substances such as coenzyme, glutathione or ferrites are essential for the reduction of small molecular compounds and the detoxification of heavy metals and exogenous substances. In recent years, due to the application of large amounts of nitrogen fertilizer and the limitation of the emission of sulfur containing waste gas, the lack of sulfur in soil and soil will be a factor restricting agricultural production. The study of sulfur absorption, transport and assimilation is clearer. The mechanism of response to sulfur deficiency in Arabidopsis is also clear, and the response of rice to sulfur deficiency is less reported. Therefore, the two week old rice seedlings were treated with sulfur deficiency stress, and the effects of sulfur deficiency stress on the growth and biochemical level of rice plants were observed. A comparative analysis was made between the normal growth and the transcriptional group of rice leaves and roots for one week without sulfur stress. The response genes of sulfur deficiency stress in rice were found, and the differential expression analysis was carried out to achieve the purpose of exploring the regulation mechanism of gene expression related to the deficiency of sulfur stress in rice. The specific results are as follows: (1) growth and biochemical levels The results showed that the POD, CAT, SOD enzyme activity and chlorophyll content began to decrease significantly in rice seedlings with two weeks of sulfur deficiency stress, and the content of glutathione was decreased at the beginning of third weeks. Compared with trees and other plants, rice was not sensitive to sulfur deficiency stress. (2) 20705 genes were detected in the control group, 31200 genes were detected in the control group and 29964 genes were detected in the control group, and 31050 genes were detected in the treatment group. A number of genes were induced in rice seedlings. The differential expression genes were detected by NOISeq technology. 866 differentially expressed genes were screened in the leaves. 11 genes were down and 855 genes were up-regulated. There were 289 differentially expressed genes in the root, 28 genes, 261 bases due to up regulation, and the common differentially expressed genes in the roots and leaves. 13 differentially expressed genes were verified by real time fluorescence quantitative PCR, and the results similar to RNA-seq were obtained to demonstrate the reliability of this high throughput sequencing. (3) the KEGG of differentially expressed genes and the significant enrichment analysis of Pathway showed that the genes related to the response of oxidative stress response to large quantities under sulfur deficiency stress occurred The differentially expressed genes are mainly related to metabolic pathways such as nucleic acid metabolism and signal transduction. The differentially expressed genes in the root are mainly related to various metabolic pathways such as energy metabolism and signal transduction. (4) the regulation of sulfur transporter in sulfur transport under sulfur stress is similar to that of Arabidopsis thaliana, indicating that rice is similar to Arabidopsis. To regulate the redistribution of sulfate; like Arabidopsis thaliana, the target genes of ATPS, APR, MSA1, SDI1, SAM and miR395 in the sulfur assimilation pathway in rice are also regulated in varying degrees. However, the lack of sulfur stress in Arabidopsis can lead to the decline in the expression level of the target gene ATPS1, ATPS3, ATPS4 of the miR395, but the expression of ATPS in rice is induced, It is likely that the regulation mechanism of mono cotyledonous and dicotyledonous plants is different. At the same time, it is predicted that Os09g27660 is probably the target gene of miR395. (5) analysis of differential expression transcription factors in roots and leaves shows that there are 21 differentially expressed transcription factors in leaves and 4 differentially expressed transcription factors in the roots, and the number of differentially expressed transcriptional factors in leaves is significantly higher At the root, these transcription factors are important for regulating the expression of sulfur deficient response genes and plant hormone metabolism. At the same time, it is found that the expression of multiple miRNA target genes changes obviously, indicating that other miRNA except miR395 may also participate in the response to sulfur deficiency stress. This study shows that the stress deficiency can promote the growth of rice root; The mechanism of sulfur transport and the assimilation pathway of rice seedlings changed to respond to the physiological state of sulfur deficiency; the oxidative stress response was induced by sulfur deficiency stress, and a large number of response genes related to oxidative stress were regulated, and a series of differentially expressed genes related to the response to sulfur deficiency stress in rice seedlings were obtained. The further study of the response regulation network for the response to sulfur deficiency stress will provide a theoretical basis for improving the resistance to stress and yield of rice, and it is of great significance to how to manipulate the sulfur metabolism and improve the yield of rice.
【學(xué)位授予單位】：河南師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：Q945.78;S511

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 程志遠(yuǎn);李穆;石瑩;何平;何麗蓮;李富生;;利用高通量測(cè)序分析甘蔗的抗旱響應(yīng)機(jī)制[J];分子植物育種;2015年09期

2 Zhen Wang;Jie-Li Mao;Ying-Jun Zhao;Chuan-You Li;Cheng-Bin Xiang;;L-Cysteine inhibits root elongation through auxin/PLETHORA and SCR/SHR pathway in Arabidopsis thaliana[J];Journal of Integrative Plant Biology;2015年02期

3 Qing Zhao;Yu Wu;Lei Gao;Jun Ma;Chuan-You Li;Cheng-Bin Xiang;;Sulfur nutrient availability regulates root elongation by affecting root indole-3-acetic acid levels and the stem cell niche[J];Journal of Integrative Plant Biology;2014年12期

4 鄧敏捷;董焱鵬;趙振利;張曉申;范國(guó)強(qiáng);;基于Illumina高通量測(cè)序的泡桐轉(zhuǎn)錄組研究[J];林業(yè)科學(xué);2013年06期

5 張憲政;植物葉綠素含量測(cè)定——丙酮乙醇混合液法[J];遼寧農(nóng)業(yè)科學(xué);1986年03期

相關(guān)博士學(xué)位論文 前3條

1 李芳軍;利用VIGS技術(shù)研究棉花抗逆基因功能[D];中國(guó)農(nóng)業(yè)大學(xué);2014年

2 宋睿;砷脅迫下硫?qū)ν寥馈局猩檫w移的影響及其機(jī)制研究[D];河南農(nóng)業(yè)大學(xué);2014年

3 曹民杰;擬南芥硫轉(zhuǎn)運(yùn)蛋白基因SULTR3;1的功能解析和miR395對(duì)硫同化通路的調(diào)控[D];中國(guó)科學(xué)技術(shù)大學(xué);2010年

相關(guān)碩士學(xué)位論文 前3條

1 秦鐘梁;水稻ONAC055的功能分析及轉(zhuǎn)基因植株的轉(zhuǎn)錄組分析[D];重慶大學(xué);2016年

2 董婷;水稻硫酸鹽轉(zhuǎn)運(yùn)蛋白OsSultr1；1基因功能的研究[D];山東大學(xué);2010年

3 劉俊超;水稻硫酸鹽轉(zhuǎn)運(yùn)蛋白OsSultr2；1的表達(dá)與功能研究[D];山東大學(xué);2009年

，

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