甘蔗果糖激酶基因家族基因組演化及功能研究
發(fā)布時間：2019-05-05 10:06
【摘要】：作為全球最重要的高糖農(nóng)作物,甘蔗由于其復(fù)雜的遺傳背景,至今尚無甘蔗全基因組序列,而對蔗糖代謝的基因組學(xué)研究處于起步階段。果糖激酶作為糖代謝中一個重要的激酶,對其在甘蔗基因組學(xué)和功能研究上均為空白。本研究應(yīng)用比較基因組學(xué)手段結(jié)合項(xiàng)目組的酵母人工染色體文庫,將根據(jù)甘蔗的近緣物種高粱的果糖激酶基因來對甘蔗果糖激酶基因家族成員進(jìn)行鑒定,在甘蔗中鑒定出果糖激酶基因家族7個成員,命名為ScFRK1-ScFRKScFRK7,并對這七個成員進(jìn)行單倍型基因結(jié)構(gòu)注釋和單倍型Ka/Ks分析。應(yīng)用甘蔗近緣屬物種高粱、玉米以及單子葉模式植物水稻,雙子葉代表性植物西紅柿、擬南芥、葡萄和馬鈴薯共8個物種的果糖激酶基因家族共71個果糖激酶基因構(gòu)建一個系統(tǒng)演化圖和結(jié)構(gòu)注釋,研究甘蔗與植物果糖激酶基因家族的系統(tǒng)演化,結(jié)果顯示甘蔗果糖激酶基因家族在單雙子葉植物分化之前已存在,屬于比較古老的基因家族。果糖激酶基因家族經(jīng)過三次的基因組復(fù)制事件后形成7個家族成員,其中最古老的基因是ScFRK6和ScFRK7,其次是ScFRK4,最后ScFRK1、ScFRK2、ScFRK3和ScFRK5。分析甘蔗果糖激酶在四個甘蔗品種的不同生長周期以及在不同激素處理下,果糖激酶基因家族表達(dá)量的差異,結(jié)果顯示ScFRK1和ScFRK5是主效基因,在不同生長周期和不同激素處里下,甘蔗果糖激酶的表達(dá)模式不一樣。其中ScFRK1表達(dá)量最高,而ScFRK5次之,SsFRK7是這個基因家族中最低的表達(dá)成員,這說明甘蔗果糖激酶家族基因SsFRK1可能是主效基因,相比其他激素脅迫,ScFRK4在GA脅迫下表達(dá)量增加明顯,推測ScFRK4在甘蔗發(fā)芽、分蘗過程中起比較重要的作用。借助模式植物擬南芥,將果糖激酶基因?qū)胝系綌M南芥突變體和野生型中,觀察擬南芥表型的變化,因?yàn)楣羌っ笇χ参锞S管組織和糖代謝有比較重要的作用,因此主要看根莖長度的變化,發(fā)現(xiàn)果糖激酶ScFRK1和ScFRK5對擬南芥根莖的生長分別有抑制和促進(jìn)的作用,具體的作用機(jī)制則需要后續(xù)進(jìn)一步的研究探討。
[Abstract]:As the most important high sugar crop in the world, sugarcane has no whole genome sequence due to its complicated genetic background, but the genomics research on sucrose metabolism is still in its infancy. Fructose kinase, as an important kinase in sugar metabolism, is blank in genomics and function research of sugarcane. In this study, combining comparative genomics with yeast artificial chromosome library, members of sugarcane fructose kinase gene family were identified according to the fructose kinase gene of sorghum, a closely related species of sugarcane. Seven members of the fructose kinase gene family were identified in sugarcane and named ScFRK1-ScFRKScFRK7,. Haplotype gene structure annotation and haplotype Ka/Ks analysis were performed on the seven members. Application of sugarcane related species sorghum, maize and monocotyledon model plant rice, dicotyledon representative plant tomato, Arabidopsis thaliana, A phylogenetic map and structural annotation of 71 fructose kinase genes from eight species of grape and potato were constructed to study the phylogenetic evolution of fructose kinase gene family in sugarcane and plants. The results showed that the fructose kinase gene family of sugarcane existed before the differentiation of dicotyledonous plants and belonged to the older gene family. The fructokinase gene family formed seven family members after three genomes replication events, the oldest of which was ScFRK6 and ScFRK7, followed by ScFRK4, ScFRK1,ScFRK2,ScFRK3 and ScFRK5.. The differences of the expression of fructose kinase gene family in four sugarcane varieties were analyzed. The results showed that ScFRK1 and ScFRK5 were the major genes in different growth cycles and different hormone treatments, and the results showed that fructose kinase gene was the main gene in different growth cycles and different hormone locations, and the expression of fructose kinase gene family was different in different growth cycles and different hormone treatments. The expression pattern of fructose kinase in sugarcane is different. The expression of ScFRK1 was the highest, followed by ScFRK5, and SsFRK7 was the lowest expressed member of this gene family. This indicated that SsFRK1 gene of sugarcane fructose kinase family might be the major gene. Compared with other hormone stress, the expression level of ScFRK4 under GA stress was significantly increased. It is suggested that ScFRK4 plays an important role in sugarcane germination and tillering. With the help of the model plant Arabidopsis thaliana, the fructose kinase gene was introduced into Arabidopsis mutants and wild type, and the phenotypic changes of Arabidopsis thaliana were observed because of the important role of fructose kinase in vascular tissue and glucose metabolism in plants. Therefore, it was found that fructose kinase ScFRK1 and ScFRK5 could inhibit and promote the growth of Arabidopsis thaliana rhizome respectively, and the specific mechanism needed to be further studied.
【學(xué)位授予單位】：福建師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S566.1;Q943.2
[Abstract]:As the most important high sugar crop in the world, sugarcane has no whole genome sequence due to its complicated genetic background, but the genomics research on sucrose metabolism is still in its infancy. Fructose kinase, as an important kinase in sugar metabolism, is blank in genomics and function research of sugarcane. In this study, combining comparative genomics with yeast artificial chromosome library, members of sugarcane fructose kinase gene family were identified according to the fructose kinase gene of sorghum, a closely related species of sugarcane. Seven members of the fructose kinase gene family were identified in sugarcane and named ScFRK1-ScFRKScFRK7,. Haplotype gene structure annotation and haplotype Ka/Ks analysis were performed on the seven members. Application of sugarcane related species sorghum, maize and monocotyledon model plant rice, dicotyledon representative plant tomato, Arabidopsis thaliana, A phylogenetic map and structural annotation of 71 fructose kinase genes from eight species of grape and potato were constructed to study the phylogenetic evolution of fructose kinase gene family in sugarcane and plants. The results showed that the fructose kinase gene family of sugarcane existed before the differentiation of dicotyledonous plants and belonged to the older gene family. The fructokinase gene family formed seven family members after three genomes replication events, the oldest of which was ScFRK6 and ScFRK7, followed by ScFRK4, ScFRK1,ScFRK2,ScFRK3 and ScFRK5.. The differences of the expression of fructose kinase gene family in four sugarcane varieties were analyzed. The results showed that ScFRK1 and ScFRK5 were the major genes in different growth cycles and different hormone treatments, and the results showed that fructose kinase gene was the main gene in different growth cycles and different hormone locations, and the expression of fructose kinase gene family was different in different growth cycles and different hormone treatments. The expression pattern of fructose kinase in sugarcane is different. The expression of ScFRK1 was the highest, followed by ScFRK5, and SsFRK7 was the lowest expressed member of this gene family. This indicated that SsFRK1 gene of sugarcane fructose kinase family might be the major gene. Compared with other hormone stress, the expression level of ScFRK4 under GA stress was significantly increased. It is suggested that ScFRK4 plays an important role in sugarcane germination and tillering. With the help of the model plant Arabidopsis thaliana, the fructose kinase gene was introduced into Arabidopsis mutants and wild type, and the phenotypic changes of Arabidopsis thaliana were observed because of the important role of fructose kinase in vascular tissue and glucose metabolism in plants. Therefore, it was found that fructose kinase ScFRK1 and ScFRK5 could inhibit and promote the growth of Arabidopsis thaliana rhizome respectively, and the specific mechanism needed to be further studied.
【學(xué)位授予單位】：福建師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S566.1;Q943.2
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