發(fā)布時(shí)間：2018-04-15 15:19

  本文選題：植物分枝 + 獨(dú)腳金內(nèi)酯�。� 參考：《華中農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：分枝為植物非常復(fù)雜的生理發(fā)育過程,涉及到多種生理機(jī)制,植物激素對(duì)分枝的影響是其中重要的一環(huán)。2008年發(fā)現(xiàn)了一類新型植物激素——獨(dú)腳金內(nèi)酯(strigolactone, SLs),其功能為參與調(diào)控植物地上部分分枝。當(dāng)前,關(guān)于獨(dú)腳金內(nèi)酯調(diào)控分枝的機(jī)理機(jī)制研究已取得一定進(jìn)展,在擬南芥(Arabi-dopsis thaliana)、水稻(Oryza sativa)、豌豆(Pisum sativum L.)、矮牽牛(petunia hybrida)等物種中已克隆出許多與獨(dú)腳金內(nèi)酯相關(guān)的基因。但至今獨(dú)腳金內(nèi)酯的合成和信號(hào)傳導(dǎo)的具體途徑尚不清晰,在不同物種中已克隆的基因亦有限,特別是其在木本植物中的作用機(jī)制還處于起步階段。而楊樹作為木本植物中的模式植物,探究獨(dú)腳金內(nèi)酯對(duì)其分枝機(jī)理的具影響有重要意義。本實(shí)驗(yàn)通過生物信息學(xué)方法,調(diào)取11個(gè)擬南芥與獨(dú)腳金內(nèi)酯合成及信號(hào)轉(zhuǎn)導(dǎo)相關(guān)的基因,比對(duì)出其在毛果楊(Populus trichocarpa)中對(duì)應(yīng)的同源基因,對(duì)篩選出的毛果楊獨(dú)腳金內(nèi)酯各途徑的同源基因,分析各基因的功能及表達(dá)情況,挑選出擬南芥及毛果楊獨(dú)腳金內(nèi)酯信號(hào)轉(zhuǎn)導(dǎo)途徑中MAX2的同源基因?yàn)槟康幕蜻M(jìn)行克隆。目前,實(shí)驗(yàn)部分使用Gateway方法已克隆出AtMAX2基因(AT2G42620),同時(shí)將AtMAX2基因在Col-0型擬南芥中過量表達(dá)；同時(shí),以歐洲山楊×銀白楊(Populus tre mula×P. Alba INRA clone N717 1-B4)組培苗為植物材料,將AtMAX2在717楊樹中過量表達(dá)；克隆出楊樹中MAX2的同源基因PtrMAX2c (Potri.006G068500).構(gòu)建已轉(zhuǎn)化AtMAX2到717楊,已鑒定出陽性植株；構(gòu)建已轉(zhuǎn)化PtrMAX2c基因到Col-0型擬南芥中,已得到PtrMAX2c轉(zhuǎn)AT T1代種子；經(jīng)過3代擬南芥植株的陽性苗篩選及RT-PCR表達(dá)量鑒定,已得到AtMAX2轉(zhuǎn)Col-0型擬南芥過量表達(dá)轉(zhuǎn)基因植株的純合子株系9個(gè)；通過陽性苗篩選篩選及RT-PCR表達(dá)量鑒定,篩選出At-MAX2轉(zhuǎn)717楊樹過量表達(dá)株系3個(gè)。
[Abstract]:Branching is a very complex process of plant physiological development, which involves a variety of physiological mechanisms.The effect of phytohormone on branching is an important one. In 2008, a new kind of plant hormone, strigolactone, SLS, was found, which is involved in the regulation of shoot branching of plants.At present, some progress has been made on the mechanism of the regulation of branching by monogolide. In Arabidopsis thaliana, Oryza sativaa, Pisum sativum L., petunia hybrida), many genes related to monopodide have been cloned in Arabidopsis thaliana.However, the specific pathway of the synthesis and signal transduction of goldactone is not clear, and the cloned genes in different species are limited, especially its mechanism of action in woody plants is still in its infancy.Poplar, as a model plant in woody plants, is of great significance to explore the effect of diptyrolactone on its branching mechanism.In this study, 11 genes related to the synthesis and signal transduction of goldactone in Arabidopsis thaliana were obtained by bioinformatics, and their homologous genes in Populus trichocarpa were compared.The homologous genes of MAX2 in the signal transduction pathway of Poplar monopodide from Arabidopsis thaliana and Populus tomentosa were selected for cloning by analyzing the function and expression of these genes.At present, the AtMAX2 gene AT2G42620 was cloned by Gateway method, and the AtMAX2 gene was overexpressed in Col-0 type Arabidopsis thaliana. At the same time, the plantlets of Populus tre mula 脳 P. INRA clone N7171-B4 were used as plant materials.AtMAX2 was overexpressed in 717 poplar and the homologous gene PtrMAX2c Potri.006G068500 was cloned.Positive plants were identified by construction of transformed AtMAX2 to 717 poplar, transformed PtrMAX2c gene into Col-0 type Arabidopsis thaliana, and transgenic AT T1 seeds were obtained. After 3 generations of Arabidopsis thaliana plants, the positive seedlings were screened and RT-PCR expression was identified.Nine homozygous lines of Col-0 type Arabidopsis thaliana transgenic plants with AtMAX2 were obtained, and 3 lines of At-MAX2 transferred 717 poplar were screened by screening positive seedlings and identifying RT-PCR expression.
【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】：2016
【分類號(hào)】：S792.11

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