梅花bZIP基因家族生物信息学鉴定及表达分析

发布时间：2018-04-25 22:30

  本文选题：梅花 + 基因　； 参考：《安徽农业大学》2016年硕士论文

【摘要】：梅花(Prunus mume)作为中国传统名花之一,它与兰、竹、菊并称为四君子,又与松、竹一起被称为岁寒三友。梅花基因组序列的发布为在基因组水平上鉴定梅花的优良观赏性状、抗逆相关基因提供了重要资料。在各种植物的生理抗性过程中bZIP转录因子都有比较广泛的参与。迄今为止,虽然在很多植物中关于bZIP转录因子已有很多的研究报道,但是在梅花中关于bZIP基因暂时并无研究,所以在我们针对梅花进行相关生物信息学研究,探究bZIP转录因子的生物信息学功能,具有非常大的研究意义。在研究中通过生物信息学分析鉴定出梅花基因组中的bZIP家族基因,通过进化树分析、染色体定位分析、MEME蛋白基序分析、外显子-内含子结构分析等生物信息学分析方法来预测了梅花bZIP基因家族的进化和功能;通过荧光定量PCR表达量的分析研究在非生物、激素处理下梅花bZIP基因的表达状态,同时克隆梅花相关的抗逆基因。获得的主要结果如下:1、在已知的梅花基因组中通过生物信息学分析、比对,利用Pfam在线工具验证,最终获得37个梅花bZIP基因,根据在染色体定位的结果分别将37个bZIP基因命名为PmbZIP1到PmbZIP37,氨基酸长度在117到1382之间分布,平均长度为415个氨基酸。2、通过得到的梅花和拟南芥bZIP基因家族基因的氨基酸序列进行重组建树,根据拓扑学和聚类情况,将梅花bZIP基因划分为10个亚族。3、通过对梅花bZIP基因生物信息学的各种分析结果如:进化树分析、外显子-内含子分析等表明梅花bZIP聚类在相同结构中表现的更近一些,同时也表明在进化过程中梅花bZIP基因家族比较保守。4、为了获得bZIP蛋白我们通过MEME在线网站,对其进行保守基序分析。分析的结果表明在PmbZIP基因中可以找到多个保守基序,在同一类型中,bZIP基因在基序类型和基序数目上表现出一定的相似性。同时,1、2、3、4和6号保守基序被鉴定为PmbZIP蛋白核心序列,这些序列广泛的分布于梅花bZIP基因家族中。5、梅花bZIP基因在非生物、外源激素处理表达下的结果显示梅花bZIP基因在高盐、干旱处理下有22个梅花bZIP基因的表达为上调,5个梅花bZIP基因表达为下调,3个梅花bZIP基因在高盐和干旱处理下表达均为上调;19个梅花bZIP基因在ABA处理下有表达,其中有8个梅花bZIP基因的表达为上调,11个梅花bZIP基因的表达为下调。证明此家族基因在梅花抗逆中扮演着重要的角色。6、通过生物信息学分析和表达分析的结果,采用荧光定量RT-PCR技术从梅花中克隆了PmbZIP35相关抗逆基因;PmbZIP35基因ORF全长为467bp,编码氨基酸155个。这为进一步研究PmbZIP35基因功能提供前期基础。
[Abstract]:As one of the traditional Chinese famous flowers, Prunus mumetogether with orchid, bamboo and chrysanthemum are called "four gentlemen", together with pine and bamboo, they are called "three friends of the year and cold". The release of the genomic sequence of plum blossom provides important information for identifying the fine ornamental traits and stress-resistant genes of plum blossom at the genomic level. BZIP transcription factors are widely involved in the physiological resistance of various plants. Up to now, although there have been many reports on bZIP transcription factors in many plants, but there is no research on bZIP gene in plum blossom, so we do some bioinformatics research on plum blossom. It is of great significance to explore the bioinformatics function of bZIP transcription factors. In the study, bZIP family genes were identified by bioinformatics analysis, and mame protein motifs were analyzed by evolutionary tree analysis and chromosome location analysis. The evolution and function of bZIP gene family in plum blossom were predicted by bioinformatics methods such as exon-intron structure analysis, and the expression status of bZIP gene in plum blossom was studied by fluorescence quantitative PCR analysis under abiotic and hormonal treatment. At the same time, the stress resistance genes related to plum blossom were cloned. The main results obtained are as follows: 1: 1. By bioinformatics analysis, comparison and verification with Pfam online tools, 37 bZIP genes were obtained from the known plum flower genome. According to the results of chromosome mapping, 37 bZIP genes were named PmbZIP1 to PmbZIP37, and the amino acid length was between 117 and 1382. The average length was 415 amino acids. The amino acid sequences of the bZIP gene family of plum blossom and Arabidopsis thaliana were recombined and constructed according to topology and clustering. The bZIP gene of plum blossom was divided into 10 subfamilies. The results of bioinformatics analysis of the bZIP gene of plum blossom, such as evolutionary tree analysis and exon-intron analysis, showed that the cluster of bZIP in plum blossom was more recent in the same structure. It also showed that the bZIP gene family of plum blossom was conserved. In order to obtain the bZIP protein, we analyzed the conserved motif by MEME online. The results show that several conserved motifs can be found in the PmbZIP gene, and the motif type and the number of motifs are similar in the same type. At the same time, the conserved motifs were identified as the core sequences of PmbZIP protein, which were widely distributed in the bZIP gene family of plum blossom. The bZIP gene of plum blossom was abiotic. The results of exogenous hormone treatment showed that the bZIP gene of plum blossom was high salt. Under drought treatment, the expression of bZIP gene was up-regulated in 22 plum flowers, down-regulated in 5 plum blossom bZIP genes, up-regulated in 3 plum blossom bZIP genes under high salt and drought treatments, and 19 plum blossom bZIP genes were expressed under ABA treatment. The expression of bZIP gene was up-regulated in 8 of them and down-regulated in 11 of them. It is proved that this family gene plays an important role in stress resistance of plum blossom. The results of bioinformatics analysis and expression analysis show that this family gene plays an important role in resistance to stress in plum blossom. The total length of PmbZIP35 related stress resistance gene (PmbZIP35) ORF was 467 BP, encoding 155 amino acids, which was cloned from plum blossom by fluorescence quantitative RT-PCR. This provides a preliminary basis for further study on the function of PmbZIP35 gene.
【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】：S685.17;Q943.2

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