本文選題：玉米 + 隱花色素��； 參考：《山西大學(xué)》2016年碩士論文

【摘要】：玉米(Zea may)不但是重要的糧食作物,同時(shí)也是高光效的C4模式植物。隱花色素(cryptochrome,CRY)是高等植物主要的藍(lán)光受體,與光形態(tài)建成、開花起始、種子休眠、生物產(chǎn)量等性狀的發(fā)育密切相關(guān)。模式植物擬南芥的隱花色素研究已經(jīng)較為深入,在單子葉植物中除了對水稻、小麥、高粱、大麥的CRY1a進(jìn)行研究外,玉米CRY1a一直未被研究。鑒于隱花色素參與調(diào)控植物生長發(fā)育的重要作用,開展對玉米隱花色素的研究迫在眉睫。本研究利用同源克隆的方法克隆、測序驗(yàn)證得到玉米B73兩個(gè)ZmCRY1a基因的的編碼區(qū)cDNA序列;采用實(shí)時(shí)熒光定量PCR(qRT-PCR)檢測ZmCRY1a在不同器官、光質(zhì)、光轉(zhuǎn)換及長日照、短日照條件下的表達(dá)水平;利用相關(guān)生物信息學(xué)軟件分析ZmCRY1as氨基酸序列及蛋白質(zhì)功能結(jié)構(gòu)域,并對ZmCRY1a和其他植物CRY1a蛋白的同源性進(jìn)行分析。結(jié)果發(fā)現(xiàn),ZmCRY1as與AtCRY1和OsCRY1a蛋白結(jié)構(gòu)類似,包含1個(gè)DNA photolyase結(jié)構(gòu)域、1個(gè)FAD binding 7結(jié)構(gòu)域、1個(gè)Crytochrome C結(jié)構(gòu)域;氨基酸序列同源性分析發(fā)現(xiàn),ZmCRY1as與水稻OsCRY1a同源性最高,與擬南芥、大豆等雙子葉植物的CRY1的同源性較低;ZmCRY1as在成株期葉片中表達(dá)量最高:ZwCRY1a1、ZmCRY1a2在葉中的表達(dá)量分別是其根中的52.1倍、15.3倍;相對于黑暗下,二者在各種持續(xù)光質(zhì)中的表達(dá)豐度均較高,尤其藍(lán)光和遠(yuǎn)紅光條件下,分別達(dá)到黑暗中ZmCRY1a1表達(dá)量的9.5和9.0、6.3和8.3倍;盡管是作為編碼藍(lán)光受體的基因,ZmCRY1a1和ZmCRY1a2基因的表達(dá)卻能強(qiáng)烈地響應(yīng)遠(yuǎn)紅光和紅光刺激,其轉(zhuǎn)錄最大峰值分別達(dá)到各自黑暗時(shí)的4.8和3.9、22.8和14.5倍;同樣二者也能響應(yīng)光周期處理。長日照條件下,ZmCRY1a1與ZmCRY1a2的轉(zhuǎn)錄存在差異,表現(xiàn)為ZmCRY1a1的轉(zhuǎn)錄在一個(gè)光周期內(nèi)共出現(xiàn)5個(gè)峰值,最高峰位于光照12 h,而ZwCRY1a2的轉(zhuǎn)錄只有4個(gè)峰值,最高峰位于進(jìn)入黑暗2 h時(shí);短日照條件下,兩個(gè)ZmCRY1a的表達(dá)出現(xiàn)了極其相似的模式,均在進(jìn)入黑暗后出現(xiàn)兩個(gè)最高峰,分別位于一個(gè)光周期的第18 h和第22 h時(shí),而在光照期間雖然有峰值出現(xiàn),但峰值較低。同時(shí)把ZmCRY1a1和ZmCRY1a2基因構(gòu)建到玉米過表達(dá)載體pBCXUN中,轉(zhuǎn)化到農(nóng)桿菌菌株GV3101,并侵染擬南芥,收獲到T0代種子。得出的結(jié)論是:玉米ZmCRY1as與擬南芥AtCRY1a和水稻OsCRY1a具有相似的結(jié)構(gòu);ZmCRY1a在葉中表達(dá)量最高,并且能響應(yīng)各種光質(zhì)處理,特別是遠(yuǎn)紅光;ZmCRY1as也能響應(yīng)長日照和短日照條件處理。
[Abstract]:Zea maya is not only an important food crop, but also a C4 model plant with high light efficiency. Cryptochrome CRY is the main blue light receptor in higher plants, which is closely related to the development of light morphogenesis, flowering initiation, seed dormancy and biological yield. The study of cryptic pigment in Arabidopsis thaliana has been carried out deeply. Except for the study of CRY1a of rice, wheat, sorghum and barley in monocotyledonous plants, maize CRY1a has not been studied. In view of the important role of cryptosin in regulating the growth and development of plants, it is urgent to carry out the research on maize cryptic pigment. In this study, the coding region cDNA sequence of two ZmCRY1a genes of maize B73 was cloned by homologous cloning and sequenced, and ZmCRY1a was detected by real-time fluorescence quantitative PCR qRT-PCRR in different organs, light quality, light conversion and long sunlight. The ZmCRY1as amino acid sequence and protein functional domain were analyzed by bioinformatics software, and the homology of ZmCRY1a and other plant CRY1a proteins was analyzed. The results showed that ZmCRY1as had similar structure to AtCRY1 and OsCRY1a protein, including one DNA photolyase domain, one FAD binding 7 domain and one Crytochrome C domain, and the amino acid sequence homology analysis showed that ZmCRY1as had the highest homology with rice OsCRY1a and had the highest homology with Arabidopsis thaliana. The homology of CRY1 in dicotyledonous plants, such as soybean, was lower. The highest expression of ZmCRY1as in adult leaves was: ZwCRY1a1, ZmCRY1a2 in leaves was 52.1 times or 15.3 times higher than that in roots, and the expression abundance of ZmCRY1as and ZmCRY1a2 in all kinds of persistent light quality was higher than that in dark. Especially in blue light and far red light, the expression of ZmCRY1a1 was 9.5 times and 9.0 times higher than that in dark, and the expression of ZmCRY1a1 and ZmCRY1a2 genes, which encode blue light receptor, could respond strongly to the stimulation of far red light and red light, although the expression of ZmCRY1a1 and ZmCRY1a2 genes as blue light receptor could strongly respond to the stimulation of far red light and red light, respectively. The maximum transcriptional peaks were 4.8 and 3.9 ~ 22.8 and 14.5 times of their dark ones, respectively, and they could also respond to photoperiod processing. The transcription of ZmCRY1a1 was different from that of ZmCRY1a2 under long sunlight, which showed that the transcription of ZmCRY1a1 appeared five peaks in a single photoperiod, the highest peak was at 12 h of light, while the peak of ZwCRY1a2 was only 4, and the highest peak occurred at 2 h into darkness. Under the condition of short sunlight, the expression of ZmCRY1a appeared very similar pattern, both of them appeared two peaks after dark, at the 18th and 22nd hour of one photoperiod, but the peak appeared in the period of illumination. But the peak value is lower. At the same time, ZmCRY1a1 and ZmCRY1a2 genes were constructed into maize overexpression vector pBCXUN, transformed into Agrobacterium tumefaciens GV3101, infected Arabidopsis thaliana and harvested T0 seeds. It is concluded that maize ZmCRY1as has similar structure to Arabidopsis thaliana AtCRY1a and rice OsCRY1a. ZmCRY1a has the highest expression in leaves and can respond to all kinds of light quality treatments, especially to long sunlight and short sunlight.
【學(xué)位授予單位】：山西大學(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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本文編號(hào)：1904954