本文選題：嫁接　切入點(diǎn)：生物信息學(xué)分析　出處：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：柑橘作為世界第一大果樹(shù)品種,由于具有較長(zhǎng)的童期,柑橘育種受到了嚴(yán)重阻礙。在實(shí)際生產(chǎn)上采用嫁接技術(shù)可以有效縮短童期,避免實(shí)生繁殖帶來(lái)的開(kāi)花晚、始果期遲、盛產(chǎn)期延后等現(xiàn)象。嫁接苗木的始果期可以由實(shí)生苗的6-8年縮短到3-5年,而且嫁接繁殖可以根據(jù)需要選擇不同的砧木,從而改善果實(shí)的品質(zhì)和提高果實(shí)產(chǎn)量,使同一批苗木的各項(xiàng)生理生長(zhǎng)習(xí)性高度保持一致。本實(shí)驗(yàn)將克里曼丁橘嫁接于早實(shí)枳上,用接穗的母樹(shù)克里曼丁橘和未嫁接的早實(shí)枳作為對(duì)照。將它們分別進(jìn)行轉(zhuǎn)錄組測(cè)序后,進(jìn)行生物信息學(xué)分析比較。利用接穗和砧木間特異的SNP作為標(biāo)簽對(duì)mRNA進(jìn)行標(biāo)記,從而研究嫁接后接穗和砧木間mRNA的移動(dòng)和嫁接后砧穗間基因表達(dá)變化。主要結(jié)果如下:1、根據(jù)轉(zhuǎn)錄組測(cè)序分析可知,嫁接柑橘的砧木和接穗中有基因存在差異表達(dá)。而且基因表達(dá)量的變化砧木比接穗更明顯。嫁接后的砧木和對(duì)照組相比有598個(gè)基因表達(dá)量顯著上調(diào),993個(gè)基因表達(dá)量顯著下調(diào);而接穗嫁接后和對(duì)照組相比較只有53個(gè)基因表達(dá)量顯著上調(diào),69個(gè)基因表達(dá)量顯著下調(diào)。由此可知嫁接對(duì)植株的接穗部分基因表達(dá)量的影響小于對(duì)砧木部分;2、比對(duì)分析發(fā)現(xiàn),在接穗中檢測(cè)到砧木特異的SNP有563個(gè),砧木中檢測(cè)到接穗特異的SNP有1268個(gè);接穗中檢測(cè)到砧木的In Del有3個(gè),砧木中檢測(cè)到接穗的InDel有33個(gè)。結(jié)果表明嫁接植株上的砧木和接穗間存在SNP和InDel的變化;而對(duì)于嫁接植物而言,無(wú)論是SNP還是InDel,砧木的變化總是比接穗的變化大,向下移動(dòng)的數(shù)量總是比向上移動(dòng)的數(shù)量多;3、以SNP作為標(biāo)簽,利用嫁接前后砧穗組織中mRNA的SNP變化判斷嫁接植物中mRNA的移動(dòng)。分析發(fā)現(xiàn)存在向上移動(dòng)SNP的mRNA有452個(gè),存在向下移動(dòng)SNP的mRNA有960個(gè),上下均有SNP移動(dòng)的mRNA有16個(gè);結(jié)果表明植物嫁接后在接穗和砧木間存在mRNA的移動(dòng)。而且向上移動(dòng)的mRNA比向下移動(dòng)的mRNA少,同時(shí)還存在少部分的mRNA可以在砧穗間雙向移動(dòng);4、將驗(yàn)證后存在可以移動(dòng)的mRNA的基因分別在克里曼丁橘的根、莖、葉、花中做基因表達(dá)量檢驗(yàn)。發(fā)現(xiàn)基因中mRNA的移動(dòng)與基因在不同組織中的特異表達(dá)并無(wú)明顯關(guān)系。有的基因在花或者根中表達(dá)量最高。由于基因表達(dá)可能和它參與的功能有關(guān),推斷可能有的基因是在葉片或者根部合成但在花中參與生命活動(dòng)過(guò)程,所以才出現(xiàn)這類(lèi)現(xiàn)象。
[Abstract]:Citrus breeding, as the largest fruit tree variety in the world, has been seriously hindered due to its long childhood period. Grafting technique can effectively shorten the childbearing period and avoid the late flowering and early fruiting due to seed propagation. The initial fruiting period of grafted seedlings can be shortened from 6-8 years to 3-5 years, and grafting propagation can select different rootstocks according to the need, thus improving the quality of fruit and increasing fruit yield. The height of physiological growth habits of the same batch of seedlings was kept consistent. In this experiment, Creman's blueberry was grafted on the precocious trifoliate orange, the mother tree of the scion and the ungrafted precocious trifoliate trifoliate trifoliate orange were used as the control, and they were sequenced respectively. The specific SNP between scion and rootstock was used as the label to mark mRNA. The main results were as follows: 1. According to the analysis of transcriptome sequencing, we studied the shift of mRNA between scions and rootstocks and the changes of gene expression between rootstocks and scions after grafting. There was different gene expression in grafted citrus rootstock and scion, and the change of gene expression in grafted citrus rootstock was more obvious than that in scion. 598 genes expression of grafted citrus rootstock was significantly up-regulated than that of control group, and the expression of 993 genes was down-regulated significantly. However, only 53 genes were significantly up-regulated and 69 genes were down-regulated after scion grafting compared with the control group. Therefore, the effect of grafting on the gene expression of the scion part of the plant was less than that on the rootstock part, the comparative analysis showed that the gene expression of the scion part of the grafted plant was lower than that of the control group. There were 563 rootstock specific SNP detected in scions, 1 268 scion-specific SNP in rootstocks and 3 in Del in scions. 33 InDel of scions were detected in rootstocks. The results showed that there were changes of SNP and InDel between rootstocks and scions of grafted plants, but for grafted plants, the change of rootstocks was always greater than that of scions. The number of downward movement was always more than that of upward movement. The SNP changes of mRNA in panicle tissue before and after grafting were used to judge the mRNA movement in grafted plants using SNP as label. It was found that there were 452 mRNA with SNP moving upward. There were 960 mRNA with SNP moving down and 16 mRNA with SNP moving up and down, the results showed that there was mRNA movement between scion and rootstock after grafting, and the mRNA moved up was less than that moved down. At the same time, there were a few parts of mRNA that could move between rootstocks and ears in both directions. The mRNA genes that could be moved were found in the roots, stems and leaves of Cleimanthus, respectively. It was found that the movement of mRNA in the gene was not related to the specific expression of the gene in different tissues. Some genes expressed the highest amount in the flower or root. Because the gene expression may be related to the function in which it is involved, It is inferred that some genes may be synthesized in the leaves or roots but are involved in the life process in the flower, which is the reason for this phenomenon.
【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(lèi)號(hào)】：S666

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