本文選題：毛果楊 + Portri.006G237100　； 參考：《東北林業(yè)大學(xué)》2016年碩士論文

【摘要】：環(huán)化酶是一種催化核苷三磷酸生成環(huán)核苷酸的酶,廣泛存在于動(dòng)物、植物、微生物。環(huán)化酶編碼基因呈現(xiàn)基因家族特征,在植物中存在多種類型,參與植物的多種生物學(xué)功能。研究報(bào)道,水稻的一類環(huán)化酶通過控制活性氧水平參與逆境脅迫。番茄紅素ε-環(huán)化酶基因過量表達(dá)與光合作用有關(guān),番茄紅素β-環(huán)化酶基因參與小麥β-胡蘿卜素生物合成。丙二烯環(huán)化酶基因參與作物的耐鹽性,過量表達(dá)該基因增加水稻在高鹽度環(huán)境下生物量,這種功能在小麥中也有類似報(bào)道。然而,環(huán)化酶基因在樹木中的研究信息或功能,卻報(bào)道甚少。在前期研究中我們分離一個(gè)楊樹環(huán)化酶基因Potri.006G237100,它可能參與木材形成。為了鑒定該基因的功能,取得的主要研究結(jié)果如下：毛果楊環(huán)化酶基因Potri.006G237100編碼區(qū)全長CDS序列含507個(gè)核苷酸,編碼169個(gè)氨基酸。同源性檢索顯示,擬南芥、大豆及葡萄等不同植物物種均存在其同源基因。序列對(duì)比顯示,該基因編碼蛋白的氨基酸在不同的植物物種中存在較高的保守性。半定量RT-PCR分析結(jié)果顯示,毛果楊環(huán)化酶基因Potri.006G237100在木質(zhì)部、葉柄、根組織中高豐度表達(dá),且其轉(zhuǎn)錄水平與莖木質(zhì)化程度同步。構(gòu)建Potri.006G237100-GFP融合的植物載體,在擬南芥中過量表達(dá)Potri.006G237100-GFP,用激光共聚焦顯微鏡檢測(cè)轉(zhuǎn)基因擬南芥根中GFP熒光信號(hào),結(jié)果顯示信號(hào)集中細(xì)胞質(zhì)內(nèi),在周質(zhì)區(qū)域也有少量信號(hào)分布,表明Potri.006G237100蛋白可能定位在細(xì)胞質(zhì)。通過轉(zhuǎn)化擬南芥獲得Potri.006G237100過量表達(dá)遺傳材料,組織解剖學(xué)結(jié)合化學(xué)染色分析顯示,與野生型相比過表達(dá)Potri.006G237100基因擬南芥材料出現(xiàn)木質(zhì)部細(xì)胞不規(guī)整、束間細(xì)胞壁增厚；過表達(dá)Potri.006G237100基因擬南芥材料的根長與野生型擬南芥根長相比變長。這些結(jié)果表明環(huán)化酶基因Potri.006G237100可能參與楊樹木材(次生細(xì)胞壁)形成。
[Abstract]:Cyclase is an enzyme that catalyzes the formation of cyclic nucleotides by nucleoside triphosphate, which is widely found in animals, plants and microorganisms. Cyclase coding genes show the characteristics of gene family, and there are many types in plants, which participate in many biological functions of plants. It is reported that a class of cyclase in rice participates in stress stress by controlling the level of reactive oxygen species (Ros). The overexpression of lycopene 蔚 -cyclase gene is related to photosynthesis, and lycopene 尾 -cyclase gene is involved in 尾 -carotene biosynthesis in wheat. Allylene cyclase gene is involved in the salt tolerance of crops. Overexpression of the gene increases the biomass of rice under high salinity. This function is also reported in wheat. However, the research information or function of cyclase gene in trees is rarely reported. In previous studies, we isolated a poplar cyclase gene Potri.006G237100, which may be involved in wood formation. In order to identify the function of the gene, the main results are as follows: the full-length CDS sequence of the Potri.006G237100 coding region of poplar cyclase gene contains 507 nucleotides and encodes 169 amino acids. Homology search showed that homologous genes existed in Arabidopsis thaliana, soybean and grape. Sequence comparison showed that the amino acids encoded by the gene were highly conserved in different plant species. The results of semi-quantitative RT-PCR analysis showed that poplar cyclase gene Potri.006G237100 was highly abundant in xylem, petiole and root, and its transcription level was synchronized with that of stem lignification. The plant vector of Potri.006G237100-GFP fusion was constructed, and Potri.006G237100-GFPwas overexpressed in Arabidopsis thaliana. The fluorescence signal of GFP in transgenic Arabidopsis root was detected by laser confocal microscope. The results showed that the signal was concentrated in cytoplasm and distributed in periplasmic region. It was suggested that Potri.006G237100 protein might be located in cytoplasm. In Arabidopsis thaliana (Arabidopsis thaliana), Potri.006G237100 over-expressed genetic materials were obtained. The results of histological anatomy and chemical staining showed that compared with wild-type Arabidopsis thaliana with overexpression of Potri.006G237100 gene, the xylem cells were irregular and the cell wall between bundles was thicker than that of wild-type Arabidopsis. The root length of Arabidopsis thaliana with overexpression of Potri.006G237100 gene was longer than that of wild type Arabidopsis thaliana. These results suggest that cyclase gene Potri.006G237100 may be involved in the formation of poplar wood (secondary cell wall).
【學(xué)位授予單位】：東北林業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S792.11;Q943.2

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本文編號(hào)：1829099