發(fā)布時(shí)間：2018-06-23 16:35

  本文選題：甘藍(lán)型油菜(Brassica + napus)　； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：甘藍(lán)型油菜(Brassica napus L.)是油菜三大類型之一,由甘藍(lán)(B.olerac ea CC,2n=18)與白菜(B.rapa,AA,2n=20)在大約7500年前經(jīng)過自然種間雜交再雙二倍體化形成異源多倍體。遠(yuǎn)古時(shí)代油菜的細(xì)胞學(xué)、分子遺傳學(xué)及育種研究都已經(jīng)取得了重大的研究進(jìn)展,但調(diào)控根生長發(fā)育的分子機(jī)理研究較少,有必要建立油菜根尖發(fā)育階段的轉(zhuǎn)錄組信息庫。本研究旨在于油菜根尖兩個(gè)不同發(fā)育階段:根尖致死階段和根尖正常階段,進(jìn)行轉(zhuǎn)錄組測序,分析根尖發(fā)育不同階段的表達(dá)譜。探究調(diào)控根生長發(fā)育的分子機(jī)理,可為選育優(yōu)良品質(zhì)的油菜品種提供參考信息。主要研究結(jié)果如下:1、體式顯微鏡下根尖不同發(fā)育階段的形態(tài)學(xué)觀察。根尖致死和根尖正常兩個(gè)材料在種子萌發(fā)過程中的胚發(fā)育早期,出現(xiàn)表型差異:突變體材料表型明顯,根尖先出現(xiàn)一個(gè)黑點(diǎn),并逐漸變大,最終根尖停止發(fā)育。2、本研究利用Illumina Hi Seq 2000雙末端二代測序技術(shù),獲得了44.5GB的轉(zhuǎn)錄組數(shù)據(jù),采用Hisat2軟件進(jìn)行比對(duì),結(jié)果顯示Hisat的reads比對(duì)率均在90%左右。3、本研究獲得了根尖致死和根尖正常兩個(gè)發(fā)育階段的表達(dá)譜數(shù)據(jù)庫。八個(gè)樣品共有47968個(gè)基因表達(dá)。分析表明,差異基因第一個(gè)階段,根尖致死較根尖正常上調(diào)表達(dá)1864個(gè)基因,下調(diào)表達(dá)1447個(gè)基因;第二個(gè)階段根尖致死較根尖正常上調(diào)表達(dá)425個(gè)基因,下調(diào)表達(dá)488個(gè)基因;兩個(gè)階段共同上調(diào)表達(dá)119個(gè)基因,共同下調(diào)表達(dá)53個(gè)基因。4、本研究對(duì)兩個(gè)階段的差異基因,進(jìn)行了GO功能分類、GO通路顯著性富集及KEGG pathway富集分析。兩個(gè)階段的差異基因在GO功能分類中主要分布在catalytic activity、binding、cell、organelle、cell part、metabolic process、cellular process、single-organism process、regulation of biological process、response to stimulus、localization、以及biological regulation條目上。在GO數(shù)據(jù)庫中,兩個(gè)階段共同的差異基因顯著性富集到兩個(gè)路徑,分別為nucleic acid binding transcription factor activity(GO:0001071)路徑和structural constituent of cell wall路徑。將兩個(gè)階段共同差異基因比對(duì)到KEGG數(shù)據(jù)庫中,獲得了2條顯著性代謝通路,分別為Steroidbiosynthesis(KO:ko00100)類固醇合成途徑,alpha-Linolenic acid metabolism(ko:00592)代謝途徑。5、分析發(fā)現(xiàn)有四個(gè)差異基因的GO功能注釋為cell killing,這四個(gè)基因分別為Bna A05g03400D、Bna C04g49050D、Bna A02g17520D、Bna C02g23400D。將其比對(duì)到擬南芥數(shù)據(jù)庫中,發(fā)現(xiàn)Bnaco2g23400和Bna A02g17520是同源基因,Bna A05g03400和Bna C04g49050是同源基因,且這四個(gè)基因的GO功能注釋與本研究中的表型性狀相吻合,這四個(gè)基因被列為候選基因。6、基因(Bna A02g17520D)在擬擬南芥中(ATG37870)為磷酸烯醇丙酮酸羧基酶,該酶是糖異生的關(guān)鍵酶,受MC9依賴性蛋白活性的影響是MC9的底物,具有與細(xì)胞死亡相關(guān)降解的功能。本研究中GO富集結(jié)果差異基因富集到內(nèi)質(zhì)網(wǎng)蛋白加工(Protein processing in endoplasmic reticulum Ko0441)誘導(dǎo)內(nèi)質(zhì)網(wǎng)相關(guān)性降解這條代謝途徑上,兩者相互驗(yàn)證。因此,推測該基因在油菜中可能具有導(dǎo)致細(xì)胞致死的功能。
[Abstract]:Brassica napus L. (Brassica napus) is one of the three major types of rapeseed. It has been studied greatly in the cytology, molecular genetics and breeding of rapeseed in the ancient times, by B.olerac EA CC (2n=18) and cabbage (B.rapa, AA, 2n=20) about 7500 years ago by interspecific hybridization and diploid formation of heterogenous polyploid. However, there are few studies on the molecular mechanism of regulating root growth and development. It is necessary to establish a transcriptional database for the development stage of rapeseed root tip. This study aims at two different developmental stages of rapeseed root apex: the root tip death stage and the normal stage of the root apex, and the transcriptional sequence is carried out to analyze the expression profiles at different stages of root apex development. The molecular mechanism of development can provide reference information for the selection of fine quality rapeseed varieties. The main results are as follows: 1, morphological observation of different developmental stages of root apex under the body microscope. In the early development of the embryo, two materials of root tip death and root apex were in the early stage of embryo development, and there were phenotypic differences: the phenotype of the mutant material was obvious, The root tip first appeared a black spot, and gradually became larger, finally the root tip stopped developing.2. This study obtained the 44.5GB transcriptional data using the Illumina Hi Seq 2000 double terminal two generation sequencing technology, and the Hisat2 software was compared. The results showed that the reads ratio of Hisat was about 90%.3. This study obtained the root tip death and the root apex normal two. A total of 47968 genes were expressed in eight samples. The analysis showed that the first stage of the difference gene, 1864 genes were up-regulated and 1447 genes were down regulated, and 1447 genes were downregulated in the root tip. The second stages of root tip death were up to 425 genes, 488 genes were downregulated, and two steps were downregulated. The 119 genes were up-regulated and 53 genes.4 were down regulated together. The GO function classification, the significant enrichment of the GO pathway and the KEGG pathway enrichment analysis were carried out on the difference genes of the two stages. The difference genes in the two stages were mainly distributed in the catalytic activity, binding, cell, organelle, cell, etc. The bolic process, the cellular process, the single-organism process, the regulation of biological process, the response to, and the entries. The Y (GO:0001071) path and the structural constituent of cell wall path. 2 significant metabolic pathways were obtained by comparing the two stages of the common differential gene into the KEGG database, respectively, the Steroidbiosynthesis (KO:ko00100) synthesis pathway, the alpha-Linolenic acid metabolic pathway, and the analysis of four The GO function annotation of the differential gene is cell killing, and these four genes are Bna A05g03400D, Bna C04g49050D, Bna A02g17520D and Bna C02g23400D.. The four genes are listed as candidate genes.6, and the gene (Bna A02g17520D) in Arabidopsis (ATG37870) is a phosphoenolpyruvate carboxylase, which is the key enzyme of sugar isogenesis. The effect of MC9 dependent protein activity is the sole of MC9 and has the function of degradation related to cell death. In this study, G The difference genes of the O enrichment results are enriched in the endoplasmic reticulum protein processing (Protein processing in endoplasmic reticulum Ko0441) induced endoplasmic reticulum related degradation, which can be verified by each other. Therefore, it is presumed that the gene may have the function of causing cell death in rape.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S565.4

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