稻瘟病菌去泛素化酶基因Moubp1的功能分析

发布时间：2019-05-18 01:43

【摘要】：泛素化调节是一个可逆的过程,它存在一个对立的去泛素化机制,是在去泛素化酶的作用下将泛素标记从底物蛋白上移除的过程。去泛素化酶是超级大家族,对生物体代谢平衡有重要作用,参与调控肿瘤的形成和增长、细胞的分化、染色质的转录,并对神经退行性疾病产生重要影响。酵母中去泛素化酶Ubp8具有参与调控细胞进程的重要功能,不仅能够通过SAGA复合物调控组蛋白的去泛素化来影响染色质结构,还可以去泛素化非组蛋白Snf1激酶。真菌中Snf1途径高度保守并与碳代谢阻遏相关,稻瘟病菌中Snf1参与调控致病性。为了研究稻瘟病菌中该基因的功能,我们在稻瘟病菌数据库中查找Ubp8的同源基因,得到MGG__03527,该基因编码蛋白含有保守的锌指结构和UCH结构域,故将其命名为Moubp1。通过基因敲除的方法对Moubp1基因进行了功能分析。结果发现,Moubp1突变体菌株的分生孢子梗数量显著减少、产孢量显著下降,并对分生孢子的形态建成和萌发产生影响。同时,该基因的缺失使得稻瘟病菌对水稻及大麦的致病性显著下降,对洋葱和大麦表皮的穿透和扩增能力也明显降低。荧光定量分析发现,突变体菌株的黑色素合成相关基因的表达量降低。此外,基因的缺失对病菌细胞壁的敏感性和逆境胁迫产生一定的影响。荧光显微观察表明,基因Moubp1定位于细胞质中。另外,与野生型菌株相比,突变体菌株在木糖为唯一碳源培养基上的菌落生长减缓,而对2-脱氧葡萄糖的敏感性下降。综上所述,基因Moubp1在稻瘟病菌的生长发育和致病性中发挥着重要作用,同时还可能参与了碳代谢阻遏调控。
[Abstract]:Ubiquitin regulation is a reversible process, and it has an opposite delignification mechanism, which removes ubiquitin markers from substrate proteins under the action of deubiquitin enzyme. Ubiquitin enzyme is a super large family, which plays an important role in the metabolic balance of organisms. It is involved in the regulation of tumor formation and growth, cell differentiation, chromatin transcription, and has an important impact on neurodegenerative diseases. Deubiquitin enzyme Ubp8 in yeast has an important function in regulating cell process. It can not only regulate histone deubiquitin structure through SAGA complex to affect chromatin structure, but also deubiquitin non-histone Snf1 kinase. The Snf1 pathway in fungi is highly conserved and related to the suppression of carbon metabolism. Snf1 in Magnaporthe grisea is involved in the regulation of pathogenicity. In order to study the function of this gene in Magnaporthe grisea, we searched the homologous gene of Ubp8 in the database of Magnaporthe grisea, and obtained that MGG__03527, gene coding protein contains conserved zinc finger structure and UCH domain, so it is named Moubp1.. The function of Moubp1 gene was analyzed by gene knockout. The results showed that the number of conidia pedicel and sporulation of Moubp1 mutant strain decreased significantly, and had an effect on the morphogenesis and germination of conidia. At the same time, the deletion of this gene significantly decreased the pathogenicity of Magnaporthe grisea to rice and barley, and the penetration and amplification ability to onion and barley epidermis. Fluorescence quantitative analysis showed that the expression of melanin synthesis related genes in mutant strains decreased. In addition, gene deletion has a certain effect on the sensitivity of cell wall and stress. Fluorescence microscopic observation showed that the gene Moubp1 was located in the cytoplasm. In addition, compared with the wild type strain, the colony growth of the mutant strain on xylose as the sole carbon source slowed down, but the sensitivity to 2-deoxyglucose decreased. In conclusion, gene Moubp1 plays an important role in the growth, development and pathogenicity of Magnaporthe grisea, and may also be involved in the suppression and regulation of carbon metabolism.
【学位授予单位】：福建农林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S435.111.41

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