本文選題：水稻 + 超親變異系��； 參考：《東北農(nóng)業(yè)大學》2017年碩士論文

【摘要】：谷氨酰胺合成酶(GS)活性大小或基因表達量與植物蛋白質含量高低有密切關系。啟動子作為基因轉錄水平上的重要調控元件,控制著基因表達的時間和空間特性。本研究選用籽粒蛋白含量有顯著差異的秈稻品種及粳稻品種進行盆栽試驗,分析灌漿不同時期籽粒蛋白質含量、GS活性和基因表達量及GS同工型基因啟動子序列和結構變化及其與轉錄表達量間的關系,旨在深入了解GS基因啟動子的結構特點及其與轉錄水平上的調控關系,進而為解析基因轉錄水平的分子調控機理提供理論依據(jù)。研究結果表明:蛋白質含量差異顯著的品種間雜交后代通過定向選擇可以獲得超親變異的后代。蛋白質含量高的基因型在灌漿各時期積累的蛋白質量始終高于蛋白質含量低的基因型,基因型控制籽粒蛋白質積累量的高低。灌漿不同時期秈稻品種在籽粒和功能葉片中谷氨酰胺合成酶活性均較超親變異系子代及親本高,并且籽粒蛋白質含量超親子代及親本均比蛋白質含量低的子代和親本谷氨酰胺合成酶活性高。GS同工型基因的表達存在著組織特異性,GS1;1基因和GS1;2基因在籽粒和葉片中都表達;GS1;3基因只在籽粒中、GS2基因只在葉片中大量表達。灌漿不同時期高蛋白質含量品種的表達量高于低蛋白質含量品種,Os GS同工型基因表達量與酶活性和籽粒蛋白質含量高低有密切關系。籽粒和葉片中GS酶活性與蛋白質含量在灌漿不同時期都呈正相關,籽粒GS1;1基因表達量與蛋白質含量間在灌漿前期和中期都呈正相關,在灌漿不同時期葉片GS1;1和GS2及籽粒GS1;3基因的表達量與蛋白質含量間都呈正相關,灌漿中期的葉片GS1;2基因表達量對籽粒蛋白質積累影響很大。籽粒蛋白質含量有顯著差異的品種間GS同工型基因啟動子序列的同源性高達99%以上;品種間有性雜交子代GS同工型基因的啟動子序列可發(fā)生堿基的變化,但沒有導致啟動子核心元件發(fā)生變化。在調控元件數(shù)量和功能上GS同工型基因的啟動子保守性很強,啟動子在結構和功能上不易發(fā)生遺傳變異,籽粒GS同工型基因表達量的變化受GS基因啟動子的調控影響很小。GS同工型基因啟動子結構并不完全一致,不同基因啟動子在調控元件數(shù)量上有明顯的差異,但不同品種間GS同工型基因啟動子結構非常一致,GS同工型基因的表達與否和程度與多種環(huán)境因素有關系。
[Abstract]:The activity of glutamine synthase (GSH) or the amount of gene expression were closely related to the content of plant protein. As an important regulatory element at the level of gene transcription, promoter controls the temporal and spatial characteristics of gene expression. In this study, indica and japonica rice varieties with significant difference in grain protein content were selected for pot experiment. The changes of GS activity and gene expression, the sequence and structure of GS isotype gene promoter and the relationship between GS activity and transcriptional expression were analyzed at different grain filling stages. The aim of this study was to understand the structural characteristics of GS gene promoter and its relationship with transcriptional regulation in order to provide theoretical basis for analyzing the molecular regulation mechanism of GS gene transcriptional level. The results showed that cross progenies with significant difference in protein content could be obtained by directional selection. The amount of protein accumulated by genotype with high protein content was always higher than that of genotype with low protein content at each filling stage, and genotype controlled the accumulation of protein in grain. The activity of glutamine synthase in grain and functional leaves of indica rice varieties at different stages of grain filling was higher than that of progenies and parents of superparent variant lines. Moreover, the expression of glutathione synthase isotype genes in grain protein content superparents and parents were higher than those in offspring and parents with lower protein content. There was tissue specific expression of GS1G-1 gene and GS12 gene in grain and leaf. Zhondu expressed only in grains, and GS2 gene was only expressed in leaves. The expression of OS-GS isotype gene in high protein content variety was higher than that in low protein content variety at different filling stage, which was closely related to enzyme activity and grain protein content. There was a positive correlation between GS enzyme activity and protein content in grain and leaf at different stages of grain filling, and there was a positive correlation between GS1- 1 gene expression and protein content in early and middle grain filling stage. There was a positive correlation between the protein content and the expression of GS1F-1, GS2 and GS1T3 in the leaves at different filling stages, and the expression of GS1O2 in the leaves at the middle stage of grain filling had a great effect on the protein accumulation in the grain. The homology of GS isotype gene promoter sequence among varieties with significant difference in grain protein content was over 99%, and the promoter sequence of GS isotype gene in intervarietal progeny could be changed. However, it did not cause changes in the core components of the promoter. The promoter of GS isotype gene is very conservative in the number and function of regulatory elements, and it is not easy to produce genetic variation in the structure and function of the promoter. The change of GS isotype gene expression was influenced by the regulation of GS gene promoter very little. The structure of GS isotype gene promoter was not completely consistent, and there were significant differences in the number of regulatory elements among different gene promoters. However, the promoter structure of GS isotype gene was consistent among different varieties. The expression and degree of GS isotype gene were related to many environmental factors.
【學位授予單位】：東北農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S511

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