本文選題：大豆 + 轉(zhuǎn)錄組��； 參考：《中國科學院研究生院(東北地理與農(nóng)業(yè)生態(tài)研究所)》2016年碩士論文

【摘要】：大豆是一種重要的經(jīng)濟作物,為我們?nèi)粘Ｉ钐峁┲参镉秃椭参锏鞍住Ｉ谑怯绊懘蠖巩a(chǎn)量的重要因素。E1基因是大豆生育期的主效基因,研究大豆生育期基因E1的下游基因?qū)α私釫1的作用機理具有重要理論意義。本研究通過對E1超表達植株及非轉(zhuǎn)基因植株的轉(zhuǎn)錄組進行了分析,進而利用實時熒光定量PCR技術,篩選出了差異表達基因,并分析研究了這些差異基因在不同組織中的特異性表達水平,不同生長時期表達水平,長日照(光照16h/黑暗8h)和短日照(光照12h/黑暗12h)處理的表達差異。同時構建了Glyma.15G076200轉(zhuǎn)基因表達載體。首先成功對E1超表達植株和非轉(zhuǎn)基因植株進行了轉(zhuǎn)錄組高通量測序。通過測序結果的分析,我們發(fā)現(xiàn)了283個基因在E1超表達植株和野生型之間存在顯著表達差異。這些差異基因涉及植物多種代謝通路,如:開花調(diào)控、激素合成與生長發(fā)育等。總體上來看,283個基因平均地分布于大豆基因組上。依據(jù)差異基因的功能注釋,挑選了其中92個基因進行驗證,并成功地證明了27個基因是在E1超表達植株與野生型中存在表達差異。其中21個基因的表達在E1超表達的豆植株中為下調(diào)表達,有6個基因在E1超表達植株中為上調(diào)表達。我們進一步挑選了8個基因進行具體的表達分析。在明確該8個基因的組織特異性表達、時間表達模式和長短日照表達水平后,我們初步確定了一個SPL家族基因Glyma.15G076200(GmSPL3b),兩個FUL家族基因Glyma.05G018800(GmFUL2a)和Glyma.17G081200(GmFUL2b)受到E1調(diào)控并在E1開花調(diào)控通路中可能起關鍵作用,值得進一步研究。進而擬對GmSPL3b進行進一步功能分析,以Kariyutaka的cDNA為模板,成功克隆了GmSPL3b的CDS片段,并成功導入植物表達載體pTF101.1中,并以大豆品種Williams82為受體品種進行遺傳轉(zhuǎn)化。希冀獲得GmSPL3b超表達轉(zhuǎn)基因材料來分析該基因在E1通路中的作用。
[Abstract]:Soybean is an important cash crop, providing vegetable oil and vegetable protein for our daily life. Growth period is an important factor affecting soybean yield. E1 gene is the main gene in soybean growth period. It is important to study the downstream gene of E1 gene in soybean growth period to understand the mechanism of E1. In this study, the transcriptome of E1 overexpression plants and non-transgenic plants were analyzed, and then the differentially expressed genes were screened by real-time fluorescent quantitative PCR. The specific expression levels of these differentially expressed genes in different tissues, the expression levels of these genes in different growth stages, and the differences in the expression of these differentially expressed genes in long sunlight (16h/ dark for 8 h) and short sunlight (light 12h/ darkness for 12 h) were analyzed. At the same time, Glyma. 15G076200 transgenic expression vector was constructed. First, the transcriptional high-throughput sequencing of E1 and non-transgenic plants was carried out successfully. By sequencing analysis, we found that 283 genes were significantly different between E1 overexpression plants and wild type. These differentially expressed genes are involved in many metabolic pathways, such as flowering regulation, hormone synthesis, and growth and development. On the whole, 283 genes are distributed equally in soybean genome. According to the functional annotation of the differentially expressed genes, 92 of them were selected for verification, and 27 genes were proved to be differentially expressed between the E1 overexpression plants and the wild type. The expression of 21 genes was down-regulated in E1 overexpressed soybean plants, and 6 genes were up-regulated in E1 overexpression plants. We further selected 8 genes for specific expression analysis. After determining the specific expression of the eight genes, the expression pattern of time and the level of expression in long and short days, One SPL gene Glyma.15G076200 GmSPL3bN, two FUL family genes Glyma.05G018800 (GmFUL2a) and Glyma.17G081200GmFUL2b) have been identified and may play a key role in the E1 flowering regulation pathway. The CDS fragment of GmSPL3b was cloned by using Kariyutaka cDNA as template, and was successfully introduced into plant expression vector pTF101.1, and the soybean variety Williams82 was used for genetic transformation. We hope to obtain GmSPL3b transgenic material to analyze the role of GmSPL3b in E1 pathway.
【學位授予單位】：中國科學院研究生院(東北地理與農(nóng)業(yè)生態(tài)研究所)
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S565.1
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本文編號：1999819