四种被子植物热激转录因子基因家族的分子进化研究
发布时间：2018-09-18 10:14
【摘要】：高温条件下,生物体普遍的反应是产生热激蛋白,它在针对高温对农作物减产和品质下降时起着至关重要的作用。生物体在高于正常生长温度5°C时就会迅速合成一些热激蛋白在体内,而调控这些热激蛋白和其他热激诱导因子表达的调控因子-热激转录因子(Heat shock transcription factor,HSF),对提高作物耐热性能具有重要作用。目前,在拟南芥、水稻、杨树、大白菜、玉米和苹果等模式植物中对部分的HSF基因进行了研究和报道,这为我们分析杨树、拟南芥、水稻、和葡萄四种被子植物HSF基因家族的分子进化提供了条件。本研究利用生物信息学软件和在线网站等全基因组数据库,对这四种被子植物的相关基因鉴定分析、系统进化树、基因结构和保守基序、基因复制事件、种间/种内的微共线性分析、复制时间和选择压力分析、表达谱分析以及表达模式分析等方面全面展开研究,获得主要结果如下:1.拟南芥和水稻的HSF基因分别在Arabidopsis Information Resource和Rice annotation database网站上下载,杨树和葡萄用模式植物拟南芥标准HSF蛋白序列为搜索条件,在Phytozome网站中进行搜索下载,随后通过ClustalX比对,Pfam和SMART筛选和验证,最终确定杨树有30个HSF基因,葡萄有10个HSF基因。2.对来自四个物种中的87个HSF基因,基于OD结构域不同,将其划分为A、B和C三个亚族。其中,A亚族又可以划分为A1-9九个小亚族,B亚族被划分为B1-5五个小亚族。3.对这四种被子植物中20对同源基因对的内含子/外显子进行分析,位于同一亚族的相邻基因,由于其亲缘关系较近,都有着相似的基因结构,即相同的内含子数目和外显子长度。其中,只有两对同源基因(OsHsf-08/-21 and PtHsf-16/-20)的结构不同,可能是由于内含子在长期进化过程中的获得或丢失导致的。利用MEME在线网站对HSF蛋白的保守基序进行分析,再结合Pfam和SMART对HSF的保守基序进行分析。基序1是HSF最常见的核心基序。4.通过四种被子植物内的微共线性分析,说明了HSF基因家族的扩张主要通过大规模复制事件,而并非串联复制。18组(31个)同源基因对参与了片段复制事件。其中,13对来自杨树基因组,4对来自水稻基因组,1对来自拟南芥基因组,而葡萄中并没有旁系同源关系的HSF基因区段。5.拟南芥、水稻以及葡萄的基因组中包含的HSF基因区段与杨树HSF基因的复制区段具有广泛的共线性,共形成了15组直系同源基因组。可能杨树在经历两轮远古基因复制事件后,73.3%的远古复制基因被保留下来。水稻有32%的基因被保留,拟南芥仅有9%,而葡萄的复制基因全部丢失了。6.选择压力分析表明在被子植物进化过程中,纯化作用依旧发挥着主要作用,但在个别基因部分编码区域依然存在着强烈的正向选择,这些拥有正向选择的基因可能导致物种功能的进化。7.微阵列数据分析结果显示,大多数HSF基因在不同组织中都有着明显的特异性表达。其中,7组片段复制基因对有着相似的表达模式,仅有两组(PtHsf-19/-17和PtHsf-23/-27)复制基因对的表达模式略有不同。8.利用qRT-PCR技术对杨树HSF的全基因家族进行热激处理、冷胁迫处理和氧化胁迫处理三种诱导表达模式进行分析。结果表明,大部分HSF基因都受这三种胁迫的诱导。PtHsf-10受到强烈的热激诱导表达,为此我们克隆了PtHsf-10的全长基因,构建其表达载体转化拟南芥,为进一步开展其功能的研究奠定基础。
[Abstract]:At high temperatures, the general response of organisms is the production of heat shock proteins, which play a vital role in the reduction of crop yield and quality at high temperatures. When organisms grow above the normal temperature of 5 degrees C, they rapidly synthesize some heat shock proteins in vivo and regulate the expression of these heat shock proteins and other heat shock inducible factors. Heat shock transcription factor (HSF) plays an important role in improving the heat tolerance of crops. At present, some HSF genes have been studied and reported in Arabidopsis, rice, poplar, Chinese cabbage, maize and apple model plants, which can be used to analyze the four quilts of poplar, Arabidopsis, rice and grape. Molecular evolution of the HSF gene family in plants provides the basis for this study. Using bioinformatics software and genome-wide databases such as online websites, we have identified and analyzed genes related to the four angiosperms, phylogenetic trees, gene structures and conserved motifs, gene replication events, interspecific/intraspecific microcollinearity analysis, replication time and selection. The main results are as follows: 1. The HSF genes of Arabidopsis and rice were downloaded from Arabidopsis Information Resource and Rice annotation database websites respectively, and the standard HSF protein sequences of poplar and grape model plant Arabidopsis thaliana were used as search conditions. After searching and downloading on Phytozome, through ClustalX comparison, Pfam and SMART screening and validation, 30 HSF genes were identified in poplar and 10 HSF genes were identified in grape. 2. For 87 HSF genes from four species, they were divided into three subgroups A, B and C based on OD domain. The intron/exon of 20 pairs of homologous gene pairs in the four angiosperms were analyzed. The adjacent genes in the same subgroup had similar gene structure, i.e. the same intron number and exon length, because of their close genetic relationship. The structure of the source gene (OsHsf-08/-21 and PtHsf-16/-20) is different, which may be caused by the acquisition or loss of introns during long-term evolution. The conserved motifs of HSF protein were analyzed by MEME online website, and then the conserved motifs of HSF were analyzed by Pfam and SMART. Microcollinearity analysis in angiosperms showed that the expansion of HSF gene family was mainly through large-scale replication events, not tandem replication. 18 pairs (31) of homologous genes participated in fragment replication events. The HSF gene fragments in Arabidopsis, rice and grape genomes have extensive collinearity with the replication fragments of poplar HSF gene, forming 15 groups of direct homologous genomes. It is possible that 73.3% of the ancient replication genes in poplar have been preserved after two rounds of ancient gene replication events. Thirty-two percent of the genes in rice were retained, only 9 percent in Arabidopsis, and all the replication genes in grape were lost. 6. Selection stress analysis showed that purification still played a major role in angiosperm evolution, but there were still strong positive selections in some coding regions of individual genes, and these genes with positive selections were possible. The results of microarray analysis showed that most HSF genes were expressed specifically in different tissues. Seven groups of HSF gene pairs had similar expression patterns, and only two groups (PtHsf-19/-17 and PtHsf-23/-27) had slightly different expression patterns. The results showed that most of the HSF genes were induced by these three stresses. PtHsf-10 was strongly induced by heat shock. Therefore, we cloned the full-length gene of PtHsf-10 and constructed its expression vector. Arabidopsis thaliana lays the foundation for further research on its function.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2
[Abstract]:At high temperatures, the general response of organisms is the production of heat shock proteins, which play a vital role in the reduction of crop yield and quality at high temperatures. When organisms grow above the normal temperature of 5 degrees C, they rapidly synthesize some heat shock proteins in vivo and regulate the expression of these heat shock proteins and other heat shock inducible factors. Heat shock transcription factor (HSF) plays an important role in improving the heat tolerance of crops. At present, some HSF genes have been studied and reported in Arabidopsis, rice, poplar, Chinese cabbage, maize and apple model plants, which can be used to analyze the four quilts of poplar, Arabidopsis, rice and grape. Molecular evolution of the HSF gene family in plants provides the basis for this study. Using bioinformatics software and genome-wide databases such as online websites, we have identified and analyzed genes related to the four angiosperms, phylogenetic trees, gene structures and conserved motifs, gene replication events, interspecific/intraspecific microcollinearity analysis, replication time and selection. The main results are as follows: 1. The HSF genes of Arabidopsis and rice were downloaded from Arabidopsis Information Resource and Rice annotation database websites respectively, and the standard HSF protein sequences of poplar and grape model plant Arabidopsis thaliana were used as search conditions. After searching and downloading on Phytozome, through ClustalX comparison, Pfam and SMART screening and validation, 30 HSF genes were identified in poplar and 10 HSF genes were identified in grape. 2. For 87 HSF genes from four species, they were divided into three subgroups A, B and C based on OD domain. The intron/exon of 20 pairs of homologous gene pairs in the four angiosperms were analyzed. The adjacent genes in the same subgroup had similar gene structure, i.e. the same intron number and exon length, because of their close genetic relationship. The structure of the source gene (OsHsf-08/-21 and PtHsf-16/-20) is different, which may be caused by the acquisition or loss of introns during long-term evolution. The conserved motifs of HSF protein were analyzed by MEME online website, and then the conserved motifs of HSF were analyzed by Pfam and SMART. Microcollinearity analysis in angiosperms showed that the expansion of HSF gene family was mainly through large-scale replication events, not tandem replication. 18 pairs (31) of homologous genes participated in fragment replication events. The HSF gene fragments in Arabidopsis, rice and grape genomes have extensive collinearity with the replication fragments of poplar HSF gene, forming 15 groups of direct homologous genomes. It is possible that 73.3% of the ancient replication genes in poplar have been preserved after two rounds of ancient gene replication events. Thirty-two percent of the genes in rice were retained, only 9 percent in Arabidopsis, and all the replication genes in grape were lost. 6. Selection stress analysis showed that purification still played a major role in angiosperm evolution, but there were still strong positive selections in some coding regions of individual genes, and these genes with positive selections were possible. The results of microarray analysis showed that most HSF genes were expressed specifically in different tissues. Seven groups of HSF gene pairs had similar expression patterns, and only two groups (PtHsf-19/-17 and PtHsf-23/-27) had slightly different expression patterns. The results showed that most of the HSF genes were induced by these three stresses. PtHsf-10 was strongly induced by heat shock. Therefore, we cloned the full-length gene of PtHsf-10 and constructed its expression vector. Arabidopsis thaliana lays the foundation for further research on its function.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2
【参考文献】
中国期刊全文数据库 前10条
1 陈勇兵;王燕;张海利;;番茄热激转录因子(Hsf)基因家族鉴定及表达分析[J];农业生物技术学报;2015年04期
2 方璐;程锋;武剑;王晓武;;全基因组与串联复制后白菜基因的保留[J];生物技术通报;2012年11期
3 秦丹丹;彭惠茹;倪中福;许甫超;董静;张俊成;孙其信;李梅芳;;植物热激转录因子及其与耐热性关系的研究进展[J];麦类作物学报;2012年01期
4 万丙良;g衅，
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