發(fā)布時(shí)間：2019-01-05 16:55

【摘要】：沙柳作為西北地區(qū)抗旱固沙樹(shù)種,其特有的機(jī)制使它能夠在干旱沙漠環(huán)境中生存,研究其分枝及材性發(fā)育相關(guān)遺傳機(jī)制,對(duì)促進(jìn)沙柳分子定向育種,培育枝條叢生型品系,增加土壤覆蓋面積,防止沙土流失及利用沙柳生物質(zhì)能源具有重要意義。本研究以擬南芥突變體和沙柳為植物材料,分析了擬南芥pny突變體表型變化,克隆獲得了沙柳PNY基因,進(jìn)行了生物信息學(xué)分析與組織特異性表達(dá)分析。主要實(shí)驗(yàn)結(jié)果如下：(1)研究發(fā)現(xiàn)擬南芥pny突變系發(fā)育遲緩,表型變化明顯,分枝數(shù)量與莖生葉顯著增加。擬南芥pny突變系莖解剖結(jié)構(gòu)分析發(fā)現(xiàn),莖基部次生木質(zhì)部較野生型發(fā)達(dá),細(xì)胞排列緊密,韌皮部發(fā)育及向外擴(kuò)增較快。(2)利用同源克隆技術(shù),在沙柳中克隆獲得PNY基因,對(duì)PNY蛋白進(jìn)行保守序列區(qū)域及功能結(jié)構(gòu)域分析,發(fā)現(xiàn)該蛋白含有5個(gè)保守序列區(qū)域,2個(gè)功能結(jié)構(gòu)域。進(jìn)化分析表明,沙柳與杞柳和毛果楊親緣關(guān)系較近。(3)通過(guò)qRT-PCR對(duì)PNY基因在沙柳不同組織部位進(jìn)行特異性表達(dá)分析,發(fā)現(xiàn)PNY基因在沙柳的葉腋部位表達(dá)量最高,在莖、葉、頂芽,根中表達(dá)量依次降低。研究結(jié)果表明,PNY基因不僅影響分枝發(fā)育,也影響維管系統(tǒng)發(fā)育；克隆獲得了沙柳PNY基因,該基因可能為轉(zhuǎn)錄因子；沙柳SpsPNY基因及相關(guān)植物的進(jìn)化樹(shù)分析表明木本植物間的PNY基因親緣關(guān)系更近：該基因在沙柳不同組織部位的特異性表達(dá)分析表明該基因在木本植物中可能也影響維管及分枝發(fā)育。本研究成果為進(jìn)一步分析PNY基因功能奠定了研究基礎(chǔ),對(duì)樹(shù)木分子育種工作具有重要指導(dǎo)意義。
[Abstract]:As a drought resistant and sand fixation tree species in Northwest China, Salix willow is able to survive in arid desert environment by its unique mechanism. The genetic mechanism related to branch and wood sex development is studied to promote molecular orientation breeding of Salix, and to cultivate cluster species of branches. It is of great significance to increase soil cover area, prevent sand loss and utilize biomass energy of Salix. In this study, Arabidopsis thaliana mutants and Salix were used as plant materials to analyze the surface changes of Arabidopsis thaliana pny mutants. The PNY gene was cloned and analyzed by bioinformatics and tissue specific expression. The main results were as follows: (1) it was found that the pny mutant lines of Arabidopsis thaliana were stunted, phenotypic changes were obvious, and the number of branches and stem leaves were significantly increased. The anatomic structure analysis of pny mutant of Arabidopsis thaliana showed that the secondary xylem at the base of the stem was more developed than that of the wild type, the cells were arranged closely, and the phloem developed and expanded rapidly. (2) the PNY gene was cloned from Salix willow by homologous cloning technique. The conserved region and functional domain of PNY protein were analyzed. It was found that the protein contained 5 conserved regions and 2 functional domains. The phylogenetic analysis showed that Salix was closely related to willow and poplar. (3) the specific expression of PNY gene in different tissues of Salix willow was analyzed by qRT-PCR. It was found that the expression of PNY gene was the highest in the leaf axils of Salix willow and in the stem. The expression level of leaf, terminal bud and root decreased in turn. The results showed that the PNY gene not only affected the branching development but also the vascular system development, and cloned the PNY gene of Salix wilfordii, which may be a transcription factor. Phylogenetic tree analysis of Salix SpsPNY gene and related plants showed that the PNY gene was more closely related to woody plants. The specific expression of PNY gene in different tissues of Salix wilfordii showed that the gene might also have an effect on woody plants. Vascular and branching development. The results of this study lay a foundation for further analysis of PNY gene function and have important guiding significance for tree molecular breeding.
【學(xué)位授予單位】：內(nèi)蒙古農(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)】：S793.9

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1 王文娟;擬南芥pny突變體分析及沙柳PNY基因克隆與組織特異性表達(dá)分析[D];內(nèi)蒙古農(nóng)業(yè)大學(xué);2016年

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本文編號(hào)：2402048