拟南芥CDE家族基因表达模式的研究

发布时间：2018-05-10 17:28

本文选题：DUF1336 + CDE　；参考：《山东农业大学》2016年硕士论文

【摘要】：拟南芥CDE基因家族包含5个基因(CDE1,At1g10410;CE2,At1g13970;CDE3,At1g59650;CDE4,At3g29180;CDE5,At5g39430),都包含 DUF1336 结构域,属于DUF1336基因家族。DUF1336基因家族共包含15个基因。目前对拟南芥DUF1336家族基因的研究较少,仅有关于EDR2的报导,该基因与拟南芥对白粉病的抗性有关。课题对拟南芥CDE家族基因进行了相关研究,主要结果如下:课题对DUF1336基因家族的生物信息学信息进行了分析。对DUF1336基因家族所表达蛋白质的序列进行比对分析,发现该家族基因产生的蛋白质依据其结构可以分为三类,一类包含PH结构域、START结构域和DUF1336结构域,一类基本只包含DUF1336结构域,最后一类包含DUF1336结构域和N端的一段肽链,CDE基因家族属于最后一类。通过基因序列的比对分析发现DUF1336家族基因为植物特异。多物种DNA序列比对分析发现进化中较早产生的植物中不含DUF1336家族基因,而在被子植物中,许多物种也没有DUF1336家族基因。除此之外,还对CDE基因家族的蛋白质结构使用相关软件进行了预测。蛋白质跨膜性预测分析显示CDE基因家族表达的蛋白质为非跨膜蛋白。对CDE家族基因的相关突变体进行研究,转录组学数据(AtGenExpress Visualization Tool,http://www.arabidopsis.org/)显示胁迫处理会使CDE家族基因的表达水平发生明显变化,但对CDE1、CDE4的缺失突变体和野生型对照进行氯化钠处理和甘露醇处理,结果没有发现突变体有明显的表型。在拟南芥生长的幼苗时期(7d),有莲座叶但抽薹前和植株成熟时(产生成熟果荚)这三个时期对拟南芥各个部位进行半定量RT-PCR和GUS染色,用来分析拟南芥CDE家族基因的表达模式。半定量RT-PCR的结果显示:CDE1,CDE2,CDE3,CDE4在拟南芥幼苗时期根中的表达量高于地上部,在成熟植株中根、茎、花器官中表达量较高,CDE2,CDE4在花粉中的表达明显高于其它部位;CDE5全程未检测到基因表达。GUS染色分析显示:CDE基因家族5个基因的表达有差异,但很多地方相似,CDE1,CDE2,CDE3,CDE4在拟南芥营养生长时期集中在根尖,地上部气孔、叶脉中表达,生殖生长时期地上部集中在花器官中表达;CDE5表达量较少,且该基因与气孔关系密切;同时实验中发现CDE4的GUS材料在受到机械损伤时会在附近产生较多的GUS信号,表明CDE家族基因可能与机械损伤和胁迫相关。课题通过对DUF1336基因家族生物信息学信息的分析,CDE家族基因相关突变体的研究,半定量RT-PCR及GUS染色分析CDE基因家族表达模式这些实验,获得了CDE基因家族的一些基础性信息,为以后对CDE基因家族功能的研究提供了基础。
[Abstract]:The CDE gene family of Arabidopsis thaliana contains five genes, CDE1, At1g1041010, CE2U, At1g13970, CDE3, At1g59650, CDE5, AT3G29180CDE5, At5g39430, all of which contain DUF1336 domain, and belong to the DUF1336 gene family. The DUF1336 gene family contains 15 genes. There are few studies on Arabidopsis DUF1336 family genes, only about EDR2, which is related to resistance to powdery mildew in Arabidopsis thaliana. The main results are as follows: the bioinformatics information of DUF1336 gene family was analyzed. By comparing the sequences of proteins expressed in DUF1336 gene family, it was found that the proteins produced by DUF1336 gene family could be divided into three groups according to their structures, one containing PH domain, the other containing DUF1336 domain, and the other containing only DUF1336 domain. The final family of peptide chain CDE genes containing DUF1336 domain and N terminal belongs to the last class. By the alignment analysis of gene sequences, the DUF1336 family was found to be specific to plants. Multiple species DNA sequence alignment analysis showed that there were no DUF1336 family genes in plants born earlier in evolution, while in angiosperms, many species did not have DUF1336 family genes. In addition, the protein structure of CDE gene family was predicted by software. Protein transmembrane prediction analysis showed that the proteins expressed by CDE gene family were non-transmembrane proteins. In this study of CDE family genes, transcriptome data show that stress treatment can significantly change the expression level of CDE family genes, as shown by AtGenExpress Visualization tool http: / www.arabidopsis.orgr. However, sodium chloride and mannitol treatments were used to treat the deletion mutant and wild type control of CDE1 / CDE4, and no obvious phenotype was found in the mutant. At the seedling stage of Arabidopsis thaliana, there were rosette leaves before bolting and mature fruit pods. Semi-quantitative RT-PCR and GUS staining were used to analyze the expression pattern of CDE family genes in Arabidopsis thaliana. The results of semi-quantitative RT-PCR showed that the expression of CDE2 + CDE3 CDE4 in roots of Arabidopsis thaliana seedlings was higher than that in shoots, and the expression of CDE4 in roots and stems of mature plants was higher than that in roots of Arabidopsis thaliana seedlings. The expression of CDE2CDE4 in flower organs was significantly higher than that in other parts of pollen. The results of Gus staining showed that there were differences in the expression of 5 genes in the gene family of 20% CDE gene family. But in many places, CDE1, CDE2, CDE3, CDE4 were expressed in root tip, shoot stomata and leaf vein during vegetative growth of Arabidopsis thaliana, and the expression of CDE5 in floral organs was less in shoot during reproductive growth, and the gene was closely related to stomata. At the same time, it was found that the GUS material of CDE4 produced more GUS signals in the vicinity when it was subjected to mechanical damage, indicating that the CDE family genes might be related to mechanical damage and stress. Through the analysis of bioinformatics information of DUF1336 gene family, the study of gene related mutants of DUF1336 family, semi-quantitative RT-PCR and GUS staining to analyze the expression pattern of CDE gene family, some basic information of CDE gene family were obtained. It provides a basis for the study of the function of CDE gene family in the future.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2

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