本文選題：甘藍型油菜 + 蘿卜��； 參考：《華中農業(yè)大學》2016年博士論文

【摘要】：非整倍體(aneuploids)是指在一個物種正常染色體數(2n)的基礎上個別染色體增加或丟失的個體或細胞,有種內與種間兩種類型(intra-/interspecific aneuploids)。植物對非整倍性比動物具有較大的忍耐性,特別是異源多倍體可忍耐染色體丟失。前人獲得的蕓薹屬異源四倍體種甘藍型油菜(Brassica napus L.,2n=38,AACC)的一個缺體(2n=36)表現植株急劇變矮變小、大幅提早開花,但缺失染色體的“身份”有待確定。在本研究中,將該甘藍型油菜缺體與甘藍型油菜親本雜交產生了單體,然后進行該缺體和單體與甘藍型油菜親本的葉片比較轉錄組分析,以揭示特定染色體的不同拷貝(0,1,2)對轉錄組的影響、確定與染色體丟失相關表型的分子機理。還進行了兩個甘藍型油菜—蘿卜(Raphanus sativus L.,2n=18,RR)二體附加系(2n=40,AACC+1IIRR)與甘藍型油菜和蘿卜親本的葉片比較轉錄組研究,以分析單條外源染色體對油菜基因表達的影響與互作。主要研究結果如下:1.甘藍型油菜單體與缺體的轉錄組分析利用C染色體組特異的DNA重復序列為探針進行熒光原位雜交分析,顯示缺體丟失的一對染色體屬于C染色體組。從每條染色體的特定基因表達差異進一步顯示在缺體中所丟失的染色體是甘藍型油菜的C2染色體。單體和缺體的基因表達在全基因組范圍內發(fā)生了改變,且絕大多數的差異表達基因(DEGs)是由C2染色體的不同拷貝(0,1)的反式作用效應所致,因直接由C2染色體的丟失導致的DEGs在單體和缺體中只占3.89%和8.22%。其他染色體上的上調表達基因數比下調表達基因數高,可能基因組通過提高某些基因的表達來響應C2染色體的丟失。在單體中比缺體中檢測到更多的DEGs,顯示單體的整個基因表達比缺體的受到更嚴重的擾亂。其余染色體上的基因表達受到不同的影響,可發(fā)現幾個異常調節(jié)的區(qū)域,其內有集中分布的上調或下調基因。隨著C2染色體的丟失,部分同源染色體A2的平均基因表達下降。盡管缺體與單體中DEGs數目相差很大,但GO注釋的DEGs比例非常相近,進一步表明兩個非整倍體有響應C2缺失的相似機制。與缺體較矮的株高和開花提早相符的是,缺體中與植物生長素有關的基因及控制開花有關基因的表達也發(fā)生相應改變,為與C2缺失有關的形態(tài)學差異提供了部分解釋,盡管基因表達有時空性差異。2.甘藍型油菜—蘿卜二體附加系的轉錄組研究二體附加系BNR1與甘藍型油菜(BN)間的差異表達基因數(DEGs)低于BNR2的;而與蘿卜比較時則與BNR1的DEGs高于與BNR2的。附加系與親本、兩個附加系之間在差異倍數分布上有很大差異;與甘藍型油菜相比時兩個附加系中上調及下調基因分布均勻,大部分基因為非差異表達;而與蘿卜比較則下調表達基因占了絕大部分。這些結果表明附加不同的蘿卜染色體對附加系的基因表達有不同影響。附加系與甘藍型油菜之間的差異表達基因在不同基因組及染色體上的分布存在差異。在BNR1與BN的比較中,A基因組上調表達基因和下調表達基因的基因總數相近、每條染色體上的上調與下調表達基因也非常接近。但C基因組上的下調表達基因總數為上調表達的3倍,每條染色體上的下調表達基因數都多于上調表達的基因數目,其中以C1、C3、C6最為明顯。在BNR2與BN的比較中,A基因組的上調和下調表達基因總數也接近;C基因組下調表達的總體趨勢并不明顯,但C2、C3兩條染色體的下調表達基因數仍遠高于上調表達的基因數。大部分DEGs在染色體上表現隨機分布,特別是在A染色體組上,但在一些C基因組的染色體上(C1,2,3,5,6)有上調或下調基因集中出現的大的區(qū)域。兩個附加系與甘藍型油菜之間的共同差異表達基因分析發(fā)現BNR1與BNR2間共享大約一半的DEGs(1103/2349),表明這些基因可能受非整倍化的影響較大。其中269個基因在兩個附加系中上調表達,其中一部分基因與刺激/逆境響應相關。而共同下調的693個基因功能則主要集中在細胞、代謝及合成進程等。比較兩個附加系與甘藍型油菜間A與C基因組的部分同源基因的表達水平,在甘藍型油菜中有3867個基因對表現部分同源表達的偏向性(1846對偏向A基因組、2021對偏向C基因組)。但兩個附加系均比甘藍型油菜親本顯現更多的偏向性表達,表明附加蘿卜染色體后可增加甘藍型油菜中A與C基因組的表達偏向性。甘藍型油菜中的大多數偏向性表達在兩個附加系中均得以保持,也存在只在兩個附加系間共有的大量基因對,可能由外源染色體所引起。
[Abstract]:Aneuploidy (aneuploids) refers to individual or lost individuals or cells on the basis of the number of normal chromosomes (2n) of a species. There are two types of intraspecific and interspecific (intra-/interspecific aneuploids). Plants have greater tolerance to aneuploidy than animals, especially heterologous polyploidy can endure chromosome loss. One of the Brassica napus L., 2n=38, AACC of Brassica species (2n=36) obtained from Brassica species (2n=36) showed a sharp decrease in the plant and a large early flowering, but the "identity" of the missing chromosome remains to be determined. In this study, a single body was produced by the hybridization of the Brassica oleracea and Brassica napus. The leaf comparative transcriptional analysis of the deficient and monosomic and Brassica napus parents was carried out to reveal the effect of different copies of the specific chromosomes (0,1,2) on the transcriptional group and to determine the molecular mechanism of the phenotype associated with the chromosome loss. Two Brassica Raphanus (Raphanus sativus L., 2n=18, RR) was also carried out, and the 2n=40, AACC+1 (AACC+1) was also carried out. IIRR) study on the leaf comparative transcriptional group of Brassica napus and Brassica napus and radish parents to analyze the influence and interaction of single exogenous chromosome on rapeseed gene expression. The main results are as follows: 1. the transcriptional group of Brassica napus monosome and lack of body analysis using the specific DNA duplication sequence of C chromosome group as the probe for fluorescence in situ hybridization analysis, A pair of chromosomes showing missing body loss is a group of C chromosomes. The difference in the expression of specific genes from each chromosome further shows that the lost chromosomes in the missing body are the C2 chromosome of Brassica napus. The gene expression of the monomer and the missing body has changed in the whole genome, and the vast majority of the differential expression genes (DEGs) are It is caused by the trans action effect of different copies of the C2 chromosome (0,1). The number of up regulated genes on 3.89% and other chromosomes in the monomer and the other chromosomes is higher than that of the other chromosomes in the monomer and the missing body, which is caused by the loss of the C2 chromosome directly. The genome may respond to the loss of the C2 chromosome by increasing the expression of some genes. More DEGs was detected in the monomer than in the deficient body, indicating that the whole gene expression of the monomer was more severely disturbed than the missing body. The gene expression on the other chromosomes was affected differently, and several abnormal regions were found, with a centralized or down regulated basis. With the loss of the C2 chromosome, partial homologous staining was found. The average gene expression of the body A2 decreased. Although the number of DEGs in the deficient body was very different from the monomer in the monomer, the DEGs ratio of the GO annotation was very similar. It further indicated that the two aneuploidy had a similar mechanism to respond to the deletion of C2. The expression of the gene was also changed to provide a partial explanation for the morphological differences related to the C2 deletion. Although the gene expression has temporal and spatial differences, the transcriptional group of the.2. Brassica napus two body appended line studies the number of differentially expressed genes (DEGs) between the two body addition line BNR1 and the Brassica napus (BN) lower than that of the BNR2; and compared with the radish. While the DEGs of BNR1 was higher than that of BNR2, there was a great difference in the distribution of the difference multiplier between the additional lines and the parents and the two additional lines. When compared with the Brassica napus, the up-regulated and down regulated genes were evenly distributed, and most of the genes were undifferentiated. The results showed that the chromosomes of the additional radish had different effects on the gene expression of the additional lines. The distribution of differentially expressed genes between the additional lines and the Brassica napus was different in different genomes and chromosomes. In the comparison of the BNR1 and the BN, the total number of up-regulated and down regulated genes of the A genome was similar, and each stain was stained. The up-regulated and down-regulated genes on the body were also very close, but the number of down regulated genes on the C genome was 3 times up to up, and the number of down regulated genes on each chromosome was more than the number of up-regulated genes, of which C1, C3, and C6 were most obvious. In the ratio of BNR2 to BN, the A genome up-regulated and downregulated the total number of genes. The overall trend of down regulation of C genome is not obvious, but the number of down regulated genes in C2, C3 two chromosomes is still far higher than the number of up-regulated genes. Most of the DEGs is randomly distributed on the chromosomes, especially on the A chromosome group, but in some C genome chromosomes (C1,2,3,5,6) there are up or down gene concentration. The common differentially expressed gene analysis between two additional lines and Brassica napus found that BNR1 and BNR2 shared about half of DEGs (1103/2349), indicating that these genes may be affected by aneuploidy. 269 of them were expressed in two additional lines, some of which were associated with irritation / adversity. The 693 gene functions were mainly concentrated in cell, metabolism and synthesis process, and the expression level of the partial homologous genes between the A and C genomes between the two additional lines and Brassica napus was compared, and 3867 genes in the Brassica napus were biased (1846 pairs of A genomes, 2021 pairs). But the two additional lines were more biased than the Brassica napus parents, indicating that the added radish chromosomes could increase the expression bias of the A and C genomes in Brassica napus. Most of the biased expressions in Brassica napus were maintained in two additional lines and only between two additional lines. A large number of gene pairs may be caused by foreign chromosomes.

【學位授予單位】：華中農業(yè)大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S565.4

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