干旱胁迫对不同种（品种）泡桐基因表达的影响

发布时间：2018-05-06 10:48

本文选题：泡桐 + 干旱　；参考：《河南农业大学》2016年硕士论文

【摘要】：泡桐原产我国,是重要的速生用材和绿化树种,具有重要的经济和生态价值。我国泡桐种质资源丰富,分布广泛,但种间关系异常复杂,目前将泡桐属分为9种2变种和众多变异类型。为了了解不同种间泡桐优良性状的分子机制,以及他们之间的亲缘关系。本研究以白花泡桐(Paulownia fortunei)二倍体(PF2)与四倍体(PF4)、毛泡桐(Paulownia tomentosa)二倍体(PT2)与四倍体(PT4)、南方泡桐(Paulownia australis)二倍体(PA2)与四倍体(PA4)和豫杂一号泡桐(Paulownia tomentosa×Paulownia fortunei)二倍体(PTF2)与四倍体(PTF4)以及他们相应的干旱处理苗为材料,利用高通量转录组测序、小RNA测序等手段,从mRNA和miRNA水平综合分析不同种间泡桐优良性状形成的分子机制,同时在不同分子水平上对他们的遗传关系进行分析。主要结果如下:(1)本研究基于Illumina/Solexa Genome Analyzer IIx平台的转录组测序技术,将获得序列与白花泡桐基因组进行比对,基因组序列组装获得的序列结果长短相对于转录组更长,降低了计算强度且提高了注释的准确性。采用基因组为背景的转录组序列对16个样品的转录组以及miRNA进行鉴定,对4个种泡桐二、四倍体以及相应干旱处理的共有和特有基因以及miRNA进行统计分析。通过转录组分析,从16个样品中共获得27178个基因,其中:4个二倍体泡桐样品中共有25674个基因,4个二倍体干旱处理泡桐样品中共有25350个基因,4个四倍体泡桐样品中共有25821个基因,4个四倍体干旱处理泡桐样品中共有25439个基因。通过miRNA分析,从16个样品共获得517个miRNA,其中:4个二倍体泡桐样品中共有364个miRNA,4个二倍体干旱处理泡桐样品中共有298个miRNA,4个四倍体泡桐样品中共有297个miRNA,4个四倍体干旱处理泡桐样品中共有369个miRNA。在16个样品二、四倍体干旱胁迫的8组比较中,获得同时上调的差异基因127个,在这8个比较中同时下调的差异基因556个。(2)本研究中,从16个样品中共获得27178个基因,其中共表达的有17626个,这些基因在泡桐生长发育过程普遍存在,通过KEGG数据库对这些共同表达的基因进行富集分析,获得128条pathway,其中代谢途径的前三位是RNA降解、嘌呤代谢、剪切酶体。同时分别获得4组样品间共有和特有基因和miRNA。研究发现,4个种二、四倍体泡桐在响应干旱胁迫基因参与的代谢通路主要集中在:丙酮酸代谢、甘油酯代谢、半乳糖代谢、赖氨酸降解、抗坏血酸代谢、光合作用、光合器官中碳固定、光合作用-天线蛋白和次生代谢物合成等代谢通路。充分说明干旱胁迫对四种泡桐基因表达变化以及调控相关基因的影响,为我们更加深刻的了解干旱胁迫对泡桐的影响提供了更多的支持。在干旱胁迫下,在4个种二、四倍体泡桐与干旱处理比较中,虽然有很多共同表达的基因,但是每个泡桐种也都存在单独表达的基因,通过KEGG对4个泡桐种单独表达的基因的富集分析,我们发现,这些单独存在的基因在他们富集的代谢通路有很大相关性,同时也存在一定差异。说明4个泡桐种虽然存在单独存在的基因,但是这些基因在响应干旱胁迫的同时,他们之间还是有很大的密切相关性。(3)从结果来看,有9552个基因以及422个miRNA用于4个泡桐种间的遗传分析。采用转录组以及miRNA的表达变化来对不同种间泡桐进行遗传相似系数的计算以及亲缘关系图的绘制,数据量大,序列精确,结果更加准确,通过对他们遗传相似系数的计算我们发现不同组的比较中他们的遗传相似系数虽然发生一定变化,但是他们之间的亲缘关系比较明确,如果将四个泡桐种分成两组,白花泡桐、豫杂泡桐和南方泡桐一组,毛泡桐则单独一组。
[Abstract]:Paulownia is an important fast-growing and greening tree species, which has important economic and ecological value. Paulownia germplasm resources in China are rich and widely distributed, but the relationship between species is very complex. At present, Paulownia is divided into 9 kinds of 2 varieties and many types of variation. In this study, the diploid of Paulownia Paulownia (Paulownia fortunei) (PF2) and tetraploid (PF4), Paulownia tomentosa diploid (PT2) and tetraploid (PT4), Paulownia (Paulownia australis) in southern Paulownia (PA2) and tetraploid (PA4) and Henan No.1 Paulownia Paulownia diploid were used in this study. With the tetraploid (PTF4) and their corresponding drought treated seedlings as materials, using high throughput transcriptome sequencing and small RNA sequencing, the molecular mechanism of the formation of Paulownia traits in different species was analyzed from mRNA and miRNA levels, and their genetic relationships were analyzed at different molecular levels. The main results were as follows: (1) this study The sequence is compared with the genome of Paulownia Paulownia based on the Illumina/Solexa Genome Analyzer IIx platform. The sequence results obtained by the genome sequence assembly are longer than those of the transcriptional group, which reduces the calculation intensity and improves the accuracy of the annotation. The sequence of the transcriptome using the genome as the background is 16 The transcriptional group and miRNA of the samples were identified, and the common and endemic genes and miRNA of the 4 Paulownia two, tetraploid and corresponding droughts were statistically analyzed. Through the transcriptional analysis, 27178 genes were obtained from 16 samples, of which 4 diploid Paulownia samples had 25674 genes and 4 diploid droughts. There are 25350 genes in Paulownia samples and 25821 genes in 4 tetraploid Paulownia samples. There are 25439 genes in the 4 tetraploid Paulownia samples. 517 miRNA were obtained from 16 samples by miRNA analysis. Among them, 4 diploid Paulownia samples were 364 miRNA, 4 diploid Paulownia samples were treated by 298. MiRNA, 4 tetraploid Paulownia samples have 297 miRNA, 4 tetraploid drought treated Paulownia samples have 369 miRNA. in 8 groups of two and tetraploid drought stress, and 127 different genes are up to be up-regulated at the same time. 556 (2) (556) in this study, from 16 samples. 27178 genes were obtained, of which 17626 were co expressed. These genes were ubiquitous in the growth and development of Paulownia. These genes were enriched and analyzed by KEGG database, and 128 pathway were obtained. The first three of the metabolic pathways were RNA degradation, purine metabolites, and shear enzymes. At the same time, 4 groups of samples were obtained. And endemic genes and miRNA. studies found that the metabolic pathways involved in 4 species of two and tetraploid Paulownia in response to drought stress genes mainly focus on pyruvic metabolism, glyceride metabolism, galactose metabolism, lysine degradation, ascorbic acid metabolism, photosynthesis, carbon fixation in photosynthetic organs, photosynthesis - antenna protein and secondary metabolite synthesis. In order to understand the effect of drought stress on four Paulownia gene expression and the regulation of related genes, it provides more support for our understanding of the effect of drought stress on Paulownia. Under drought stress, there are a lot of common bases in the comparison of 4 species two, tetraploid Paulownia and drought treatment. However, each Paulownia species also has a single expressed gene. Through the enrichment analysis of the genes expressed separately by 4 Paulownia species by KEGG, we found that these isolated genes are closely related to their enriched metabolic pathways, and there are certain differences. The 4 Paulownia species, although there is a single gene, exist, but there is a single gene that exists in the Paulownia species, but there is a single gene in the Paulownia species. These genes were closely related to drought stress. (3) from the results, 9552 genes and 422 miRNA were used in the genetic analysis of 4 Paulownia species. The genetic similarity coefficient of Paulownia in different species of Paulownia was calculated by the transcriptional group and the changes of miRNA expression. The data are large, the sequence is accurate and the results are more accurate. Through the calculation of their genetic similarity coefficient, we found that their genetic similarity coefficients in different groups have changed, but their relationship is clear. If four Paulownia species are divided into two groups, Paulownia Paulownia, Paulownia and South Paulownia is a group, and Paulownia is a separate group.

【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S792.43

【参考文献】