本文選題：小麥 + MADS-box　； 參考：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：小麥是我國乃至全世界重要的糧食作物,它占到全球人口主食的40%以上。合適的抽穗時間對于小麥產(chǎn)量形成至關(guān)重要,研究小麥抽穗時間的調(diào)控基因可以為其遺傳改良提供重要的參考資料。本研究從小麥中克隆了一個MADS-box基因,依據(jù)參考序列比對結(jié)果,命名為Triticum aestivum L.AGAMOUS-Like 32(TaAGL32),對其表達模式和生物學(xué)功能進行了初步探究。主要結(jié)果如下:(1)TaAGL32基因有三個拷貝,分別定位于7A、7B、7D染色體,基因組序列全長分別是1908bp、1912bp和1790bp�；蚪Y(jié)構(gòu)分析顯示三個基因含有七個外顯子和六個內(nèi)含子。相應(yīng)的CDS長度均為693bp,三者之間僅有22個SNP差異,基因可編碼230個氨基酸。(2)TaAGL32蛋白均含有兩個保守的結(jié)構(gòu)域,分別是MEF2-like結(jié)構(gòu)域和K-Box結(jié)構(gòu)域,屬于典型的MIKC型蛋白。進化樹分析表明,TaAGL32基因與其祖先種相比具有進化保守性。TaAGL32蛋白亞細胞定位分析顯示其定位于細胞核中。(3)qRT-PCR結(jié)果顯示,TaAGL32基因在多個組織中均有表達,其中在根與莖中的表達量高,在小穗和小花中也有較高的表達。晝夜節(jié)律分析結(jié)果顯示TaAGL32基因表現(xiàn)出晝夜節(jié)律性,即TaAGL32基因的表達隨著光照時間增加而逐漸提高,并在光照12h后達到表達峰值,繼續(xù)光照反而表達下調(diào),并在光照結(jié)束時到達最低。(4)利用RNA干擾策略降低了TaAGL32基因在小麥中的表達。結(jié)果發(fā)現(xiàn),轉(zhuǎn)基因小麥在長日照條件下延遲抽穗5-7d,而在自然條件下轉(zhuǎn)基因小麥與對照相比晚抽穗3-4d。該基因的四倍體小麥突變體也表現(xiàn)出晚抽穗的表型。這些結(jié)果表明TaAGL32基因在小麥抽穗過程可能起到調(diào)控作用。(5)對轉(zhuǎn)基因小麥其他農(nóng)藝性狀的統(tǒng)計顯示,轉(zhuǎn)基因株系株高低于對照材料,兩個株系分別比對照矮約4.1cm和2.2cm,差異顯著;轉(zhuǎn)基因株系的穗長也有顯著地下降,與對照相比較短約1.5cm;小穗數(shù)目和千粒重有不同程度的降低,但是差異不明顯。
[Abstract]:Wheat is an important food crop in China and the world. It accounts for more than 40% of the staple food in the world. The suitable heading time is very important for wheat yield formation, and the study of the regulating genes of wheat heading time can provide important references for its genetic improvement. In this study, a MADS-box gene was cloned from wheat and named Triticum aestivum L.AGAMOUS-Like 32 TaAGL32 according to the results of reference sequence alignment. The expression pattern and biological function of MADS-box gene were preliminarily studied. The main results are as follows: there are three copies of the TaAGL32 gene, which are located on chromosome 7A 7BN 7D respectively. The genome sequences are 1908 BP, 1912 BP and 1790 BP, respectively. Genetic analysis showed that three genes contained seven exons and six introns. The corresponding length of CDS is 693bp. there are only 22 SNP differences among them. The gene can encode 230 amino acids. TaAGL32 protein contains two conserved domains, namely MEF2-like domain and K-box domain, which belong to the typical MIKC-type protein. Phylogenetic tree analysis showed that the TaAGL32 gene was evolutionarily conserved compared with its ancestors. The subcellular localization analysis of TaAGL32 protein showed that it was located in the nucleus. The results of QRT-PCR showed that the TaAGL32 gene was expressed in many tissues, especially in the roots and stems. High expression was also found in spikelets and florets. The results of circadian rhythm analysis showed that TaAGL32 gene showed circadian rhythm, that is, the expression of TaAGL32 gene gradually increased with the increase of illumination time, and reached the peak after 12 hours of illumination, and then decreased with the light exposure. At the end of illumination, TaAGL32 gene expression in wheat was reduced by RNA interference. The results showed that the late heading of transgenic wheat was 3-4 days later than that of the control under the condition of long sunshine, but the late heading of transgenic wheat was 3-4 days later than that of the control under natural conditions. The tetraploid wheat mutant of the gene also showed late heading phenotype. These results indicated that TaAGL32 gene might play a regulatory role in the heading process of wheat. The statistics of other agronomic characters of transgenic wheat showed that the plant height of transgenic lines was lower than that of the control lines, and the difference between the two lines was significant compared with the control, about 4.1cm and 2.2 cm, respectively. The panicle length of transgenic lines also decreased significantly, which was about 1.5 cm shorter than that of the control, and the number of spikelets and 1000-grain weight decreased to some extent, but the difference was not significant.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S512.1

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