大豆生育期基因E1调控开花通路中相关基因的挖掘

发布时间：2018-06-09 13:03

  本文选题：大豆 + 转录组　； 参考：《中国科学院研究生院(东北地理与农业生态研究所)》2016年硕士论文

【摘要】：大豆是一种重要的经济作物,为我们日常生活提供植物油和植物蛋白。生育期是影响大豆产量的重要因素。E1基因是大豆生育期的主效基因,研究大豆生育期基因E1的下游基因对了解E1的作用机理具有重要理论意义。本研究通过对E1超表达植株及非转基因植株的转录组进行了分析,进而利用实时荧光定量PCR技术,筛选出了差异表达基因,并分析研究了这些差异基因在不同组织中的特异性表达水平,不同生长时期表达水平,长日照(光照16h/黑暗8h)和短日照(光照12h/黑暗12h)处理的表达差异。同时构建了Glyma.15G076200转基因表达载体。首先成功对E1超表达植株和非转基因植株进行了转录组高通量测序。通过测序结果的分析,我们发现了283个基因在E1超表达植株和野生型之间存在显著表达差异。这些差异基因涉及植物多种代谢通路,如:开花调控、激素合成与生长发育等。总体上来看,283个基因平均地分布于大豆基因组上。依据差异基因的功能注释,挑选了其中92个基因进行验证,并成功地证明了27个基因是在E1超表达植株与野生型中存在表达差异。其中21个基因的表达在E1超表达的豆植株中为下调表达,有6个基因在E1超表达植株中为上调表达。我们进一步挑选了8个基因进行具体的表达分析。在明确该8个基因的组织特异性表达、时间表达模式和长短日照表达水平后,我们初步确定了一个SPL家族基因Glyma.15G076200(GmSPL3b),两个FUL家族基因Glyma.05G018800(GmFUL2a)和Glyma.17G081200(GmFUL2b)受到E1调控并在E1开花调控通路中可能起关键作用,值得进一步研究。进而拟对GmSPL3b进行进一步功能分析,以Kariyutaka的cDNA为模板,成功克隆了GmSPL3b的CDS片段,并成功导入植物表达载体pTF101.1中,并以大豆品种Williams82为受体品种进行遗传转化。希冀获得GmSPL3b超表达转基因材料来分析该基因在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.
【学位授予单位】：中国科学院研究生院(东北地理与农业生态研究所)
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S565.1
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本文编号：1999819