本文選題：水稻 + 玉米��； 參考：《中國農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：植物多肽激素是一類經(jīng)剪切后形成具有特殊功能的、成熟的多肽,長度一般為20個氨基酸左右,介導(dǎo)植物細胞間信號轉(zhuǎn)導(dǎo),并廣泛參與調(diào)控植物生長發(fā)育。目前植物中多肽激素家族主要有CLE基因家族、RGF基因家族、ERF基因家族和CEP基因家族等。本論文以糧食作物水稻和玉米為研究對象,在全基因組水平上對CEP多肽激素家族基因進行鑒定,在此基礎(chǔ)上,對同源的水稻OsCEP6.1和玉米ZmCEP1基因功能進行了轉(zhuǎn)基因功能分析,以期為進一步探討CEP多肽激素與作物生長發(fā)育的關(guān)系提供參考依據(jù)。獲得的主要研究結(jié)果如下：1.在水稻基因組中共鑒定出15個CEP多肽激素基因,定位于水稻的6條染色體上。同源進化分析結(jié)果顯示,水稻CEP家族基因可以分為2個群,且這兩個群的CEP結(jié)構(gòu)域的C-末端都存在相對保守的蛋白序列(SPGV/IGH/N)。利用Rice qTeller數(shù)據(jù)庫對其進行電子表達模式譜分析發(fā)現(xiàn),OsCEP5和OsCEP6.1在水稻的花序分生組織中表達量顯著高于其他基因。2.采用實時定量PCR的方法對OsCEP6.1基因的表達模式進行了分析,發(fā)現(xiàn)OsCEP6.1具有明顯的時空表達特異性,且能夠受到外源細胞分裂素誘導(dǎo)上調(diào)表達。外源施加人工合成的OsCEP6.1能夠顯著地抑制初生根和幼苗的生長。另外,我們還獲得了OsCEP6.1基因的超表達轉(zhuǎn)基因水稻純系,對其表型分析結(jié)果顯示,OsCEP6.1基因水稻超表達株系的株高、分蘗數(shù)目、劍葉長度、穗長、一級及二級枝梗數(shù)目、粒長、粒寬及千粒重等性狀較野生型均顯著降低。研究還發(fā)現(xiàn),轉(zhuǎn)基因株系的表型變異是由于細胞變小造成的,而細胞數(shù)目并沒有發(fā)生顯著改變。3.在玉米基因組中鑒定出12個CEP多肽激素基因,定位于玉米的8條染色體上。同源進化分析結(jié)果顯示,玉米CEP家族基因可以分為2個群。利用Maize qTeller數(shù)據(jù)庫對其電子表達模式進行分析發(fā)現(xiàn),在玉米雌穗和雄穗的花序分生組織中,ZmCEPl表達量顯著高于其他基因。4.采用實時定量PCR的方法對ZmCEPl的表達模式進行分析,發(fā)現(xiàn)ZmCEPl在玉米發(fā)育的雌穗和發(fā)育的雄穗中表達量較高,能夠受到外源細胞分裂素誘導(dǎo)上調(diào)表達。原位雜交的結(jié)果顯示,ZmCEPl主要在玉米雌穗穗軸維管束和未發(fā)育完全的小花中表達。獲得了ZmCEPl基因的擬南芥和玉米超表達轉(zhuǎn)基因株系,并進行了表型分析,發(fā)現(xiàn)超表達ZmCEPl的擬南芥植株表現(xiàn)出初生根變短和植株變小的表型。玉米超表達ZmCEPl株系的株高、穗位高、穗長、穗粗、行粒數(shù)、粒長和百粒重也較野生型均顯著降低,但是穗行數(shù)沒有顯著差異。
[Abstract]:Plant polypeptide hormone is a kind of mature polypeptide with special function, which is usually about 20 amino acids in length. It mediates plant intercellular signal transduction and is widely involved in regulating plant growth and development. At present, the polypeptide hormone family in plants mainly includes CLE gene family, CEP gene family and so on. In this paper, rice and maize were used as research objects, CEP polypeptide hormone family genes were identified at the whole genome level, and the function of OsCEP6.1 gene and maize ZmCEP1 gene of rice and maize were analyzed. So as to provide reference for further study on the relationship between CEP polypeptide hormones and crop growth and development. The main results obtained are as follows: 1. A total of 15 CEP polypeptide hormone genes were identified in rice genome and located on 6 chromosomes of rice. The results of homologous evolution analysis showed that the rice CEP family genes could be divided into two groups, and the C-terminal of the CEP domain of the two populations had a relatively conserved protein sequence, SPGV / IGH / NV. The expression of OsCEP5 and OsCEP6.1 in inflorescence meristem of rice was significantly higher than that of other genes. The expression pattern of OsCEP6.1 gene was analyzed by real-time quantitative PCR. It was found that OsCEP6.1 had obvious spatiotemporal specificity and could be up-regulated by exogenous cytokinins. Exogenous synthetic OsCEP6.1 could significantly inhibit primary rooting and seedling growth. In addition, we also obtained the pure lines of transgenic rice with OsCEP6.1 gene overexpression. The phenotypic analysis showed that the plant height, the number of tillers, the length of flag leaf, the ear length, the number of primary and secondary branches, and the grain length of OsCEP6.1 transgenic rice lines were obtained. Grain width and 1000-grain weight were significantly lower than those of wild type. The study also found that the phenotypic variation of transgenic lines was caused by the cells becoming smaller, but the number of cells did not change significantly. 3. Twelve CEP polypeptide hormone genes were identified in maize genome and located on 8 chromosomes of maize. The results of homologous evolution analysis showed that maize CEP family genes could be divided into two groups. The expression of ZmCEPl in the inflorescence meristem of female and male ears of maize was significantly higher than that of other genes. The expression pattern of ZmCEPl was analyzed by real-time quantitative PCR. It was found that the expression of ZmCEPl was higher in female ear and male ear of maize, and could be up-regulated by exogenous cytokinin. The results of in situ hybridization showed that ZmCEPl was mainly expressed in the vascular bundles of female spikes and immature florets in maize. Arabidopsis thaliana and maize super-expression transgenic lines with ZmCEPl gene were obtained and phenotypic analysis was carried out. It was found that Arabidopsis thaliana plants with overexpression of ZmCEPl had the phenotypes of shorter initial rooting and smaller plants. The plant height, ear length, ear diameter, grain number per row, grain length and 100-grain weight of ZmCEPl lines were significantly lower than those of wild type, but there was no significant difference in the number of rows per ear.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：Q943.2

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