發(fā)布時(shí)間：2018-04-25 22:30

  本文選題：梅花 + 基因　； 參考：《安徽農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：梅花(Prunus mume)作為中國傳統(tǒng)名花之一,它與蘭、竹、菊并稱為四君子,又與松、竹一起被稱為歲寒三友。梅花基因組序列的發(fā)布為在基因組水平上鑒定梅花的優(yōu)良觀賞性狀、抗逆相關(guān)基因提供了重要資料。在各種植物的生理抗性過程中bZIP轉(zhuǎn)錄因子都有比較廣泛的參與。迄今為止,雖然在很多植物中關(guān)于bZIP轉(zhuǎn)錄因子已有很多的研究報(bào)道,但是在梅花中關(guān)于bZIP基因暫時(shí)并無研究,所以在我們針對(duì)梅花進(jìn)行相關(guān)生物信息學(xué)研究,探究bZIP轉(zhuǎn)錄因子的生物信息學(xué)功能,具有非常大的研究意義。在研究中通過生物信息學(xué)分析鑒定出梅花基因組中的bZIP家族基因,通過進(jìn)化樹分析、染色體定位分析、MEME蛋白基序分析、外顯子-內(nèi)含子結(jié)構(gòu)分析等生物信息學(xué)分析方法來預(yù)測(cè)了梅花bZIP基因家族的進(jìn)化和功能;通過熒光定量PCR表達(dá)量的分析研究在非生物、激素處理下梅花bZIP基因的表達(dá)狀態(tài),同時(shí)克隆梅花相關(guān)的抗逆基因。獲得的主要結(jié)果如下:1、在已知的梅花基因組中通過生物信息學(xué)分析、比對(duì),利用Pfam在線工具驗(yàn)證,最終獲得37個(gè)梅花bZIP基因,根據(jù)在染色體定位的結(jié)果分別將37個(gè)bZIP基因命名為PmbZIP1到PmbZIP37,氨基酸長度在117到1382之間分布,平均長度為415個(gè)氨基酸。2、通過得到的梅花和擬南芥bZIP基因家族基因的氨基酸序列進(jìn)行重組建樹,根據(jù)拓?fù)鋵W(xué)和聚類情況,將梅花bZIP基因劃分為10個(gè)亞族。3、通過對(duì)梅花bZIP基因生物信息學(xué)的各種分析結(jié)果如:進(jìn)化樹分析、外顯子-內(nèi)含子分析等表明梅花bZIP聚類在相同結(jié)構(gòu)中表現(xiàn)的更近一些,同時(shí)也表明在進(jìn)化過程中梅花bZIP基因家族比較保守。4、為了獲得bZIP蛋白我們通過MEME在線網(wǎng)站,對(duì)其進(jìn)行保守基序分析。分析的結(jié)果表明在PmbZIP基因中可以找到多個(gè)保守基序,在同一類型中,bZIP基因在基序類型和基序數(shù)目上表現(xiàn)出一定的相似性。同時(shí),1、2、3、4和6號(hào)保守基序被鑒定為PmbZIP蛋白核心序列,這些序列廣泛的分布于梅花bZIP基因家族中。5、梅花bZIP基因在非生物、外源激素處理表達(dá)下的結(jié)果顯示梅花bZIP基因在高鹽、干旱處理下有22個(gè)梅花bZIP基因的表達(dá)為上調(diào),5個(gè)梅花bZIP基因表達(dá)為下調(diào),3個(gè)梅花bZIP基因在高鹽和干旱處理下表達(dá)均為上調(diào);19個(gè)梅花bZIP基因在ABA處理下有表達(dá),其中有8個(gè)梅花bZIP基因的表達(dá)為上調(diào),11個(gè)梅花bZIP基因的表達(dá)為下調(diào)。證明此家族基因在梅花抗逆中扮演著重要的角色。6、通過生物信息學(xué)分析和表達(dá)分析的結(jié)果,采用熒光定量RT-PCR技術(shù)從梅花中克隆了PmbZIP35相關(guān)抗逆基因;PmbZIP35基因ORF全長為467bp,編碼氨基酸155個(gè)。這為進(jìn)一步研究PmbZIP35基因功能提供前期基礎(chǔ)。
[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.
【學(xué)位授予單位】：安徽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S685.17;Q943.2

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本文編號(hào)：1803286