本文關(guān)鍵詞： 損傷誘導(dǎo)基因 轉(zhuǎn)基因擬南芥 棉鈴蟲 抗蟲性 胡楊 大片段DNA 突變體庫　出處：《天津大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：棉鈴蟲（Helicoverpa armigera）是世界性重要農(nóng)業(yè)害蟲，它危害范圍廣、繁殖潛力大，對(duì)環(huán)境適應(yīng)能力強(qiáng)。長期以來棉鈴蟲的防治主要是依賴化學(xué)農(nóng)藥，由于化學(xué)農(nóng)藥不合理的使用，使棉鈴蟲的抗性劇增，防治效果極不理想。同時(shí)，，大量的化學(xué)合成農(nóng)藥的使用，也向環(huán)境釋放了大量的化學(xué)合成物質(zhì)，直接或間接地對(duì)環(huán)境、生物和人類的健康構(gòu)成了極大的威脅，也成為經(jīng)濟(jì)及社會(huì)發(fā)展的重大障礙因素之一。因此，進(jìn)行生物防治是非常必要的。在本研究中，將三個(gè)雜交楊（Populus trichocarpa×美洲黑楊）基因PtdKTI5， PtdWIN4，PtdPOP3和1個(gè)白楊（Populus tremuloides Michx）基因PtKTI2分別轉(zhuǎn)入擬南芥（Arabidopsisthaliana）進(jìn)行過量表達(dá)。通過RT-PCR分析，證實(shí)了每個(gè)轉(zhuǎn)基因擬南芥品系在轉(zhuǎn)錄水平上的高表達(dá)。用這四種轉(zhuǎn)基因擬南芥喂養(yǎng)棉鈴蟲，并評(píng)估了其發(fā)育情況，體重變化以及存活率。結(jié)果表明：這四個(gè)楊樹防御相關(guān)基因?qū)γ掴徬x幼蟲和化蛹階段表現(xiàn)出不同程度的抗蟲作用，可用于植物基因工程來進(jìn)行作物改良。 林木抗逆、材性、生長發(fā)育等重要性狀多是由多基因、多條信號(hào)途徑交叉控制的復(fù)雜性狀。林木基因工程研究雖然取得了一定進(jìn)展，但目前克隆得到的單個(gè)基因在育種上的作用有限。植物經(jīng)過長期進(jìn)化，在染色體中形成了針對(duì)某性狀的自然存在的基因簇。將外源基因簇導(dǎo)入植物基因組的同一位點(diǎn)，有可能賦予受體由多基因控制的新性狀。大片段基因簇的同步插入還可以在一定程度上克服轉(zhuǎn)基因帶來的位置效應(yīng)，減少基因沉默等不良現(xiàn)象。把多個(gè)以上協(xié)同作用的基因同時(shí)轉(zhuǎn)入植物，將會(huì)獲得比單基因轉(zhuǎn)化更能獲得理想的結(jié)果。胡楊是楊柳科楊屬胡楊亞屬的一種植物，常生長在沙漠中，它耐寒、耐旱、耐鹽堿、抗風(fēng)沙，并具有很強(qiáng)的生命力。本課題將以胡楊為材料，在前期獲得的胡楊基因組大片段可轉(zhuǎn)化細(xì)菌染色體庫基礎(chǔ)上，結(jié)合大片段DNA轉(zhuǎn)化技術(shù)，獲得擬南芥突變體庫，進(jìn)行基因功能驗(yàn)證，篩選出具有潛在育種價(jià)值的關(guān)鍵基因（簇）及調(diào)控元件，分離鑒定一批具有自主知識(shí)產(chǎn)權(quán)的優(yōu)良性狀相關(guān)基因（簇）及調(diào)控元件，為進(jìn)一步通過分子育種方法培育高抗、優(yōu)質(zhì)林木新品種提供基礎(chǔ)和保障。
[Abstract]:Helicoverpa armigera (Helicoverpa armigera) is an important agricultural pest in the world. At the same time, the use of a large number of chemical synthetic pesticides also released a large number of chemical synthetic substances to the environment, directly or indirectly to the environment, Biological and human health pose a great threat and become one of the major obstacles to economic and social development. Biological control is therefore essential. Three hybrid poplar genes PtdKTI5, PtdWIN4PtdPOP3 and one Populus tremuloides Michx were transferred into Arabidopsis thaliana (Arabidopsis thaliana) for overexpression. Confirmed the high expression at the transcriptional level of each transgenic Arabidopsis strain. The four transgenic Arabidopsis strains were fed to cotton bollworm and their development was evaluated. The results showed that the four poplar defense-related genes showed different degrees of resistance to cotton bollworm larvae and pupae stage and could be used in plant genetic engineering for crop improvement. Tree stress resistance, wood properties, growth and development are complex traits controlled by many genes and multiple signal pathways. Although some progress has been made in the study of tree genetic engineering, However, the single gene cloned at present has a limited role in breeding. After a long period of evolution, plants have formed naturally existing gene clusters in the chromosomes for a certain trait. The exogenous gene clusters are introduced into the same locus of the plant genome. It is possible to endow the receptor with new traits controlled by multiple genes. Simultaneous insertion of large gene clusters can also overcome the locational effects of transgenic genes to some extent. Reduce adverse phenomena such as gene silencing. Transferring more than one synergistic gene into a plant at the same time will yield a better result than a single gene transformation. Populus euphratica is a subgenus of Populus euphratica in the willow family, which often grows in the desert. It is hardy, drought-tolerant, saline-alkali resistant, wind-resistant, and has strong vitality. In this paper, Populus euphratica will be used as a material, on the basis of the genome large fragment transmutable bacterial chromosome bank obtained in the early stage, combined with the technology of large fragment DNA transformation. Arabidopsis mutants library was obtained, gene function was verified, key genes (clusters) and regulatory elements with potential breeding value were screened, and a number of genes (clusters) and regulatory elements with independent intellectual property rights were isolated and identified. It provides the foundation and guarantee for breeding new varieties of high resistance and high quality forest by molecular breeding method.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：Q943.2;S433

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