發(fā)布時(shí)間：2018-04-19 12:22

  本文選題：中間錦雞兒 + NAC轉(zhuǎn)錄因子��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：植物在受到干旱、高溫、鹽堿、冷、蟲(chóng)害等脅迫以后,細(xì)胞會(huì)受到不同程度的損傷。中間錦雞兒具有耐寒、抗旱、耐鹽堿和貧瘠等特點(diǎn),具有良好的防風(fēng)固沙和保持水土功能。NAC轉(zhuǎn)錄因子家族是植物特有的、最大的轉(zhuǎn)錄因子家族之一,在植物應(yīng)答非生物脅迫和生長(zhǎng)發(fā)育過(guò)程中有重要的功能。本文從中間錦雞兒中克隆得到CiNAC1基因并對(duì)其進(jìn)行功能分析,具體結(jié)果如下:1.通過(guò)PCR技術(shù),克隆得到了中間錦雞兒CiNAC1基因1066 bp的cDNA全長(zhǎng)序列。生物信息學(xué)分析顯示,CiNAC1基因的開(kāi)放閱讀框ORF為921 bp,編碼306個(gè)氨基酸,推導(dǎo)的蛋白分子量為34.57 kD,等電點(diǎn)為8.35,是一種親水性蛋白,N端具有保守的NAM結(jié)構(gòu)域,具有26個(gè)磷酸化位點(diǎn)和7個(gè)糖基化位點(diǎn)。2.實(shí)時(shí)熒光定量RT-PCR檢測(cè)顯示,中間錦雞兒CiNAC1基因表達(dá)受干旱、高鹽、脫水、高pH誘導(dǎo),這表明CiNAC1基因可能與中間錦雞兒響應(yīng)逆境脅迫機(jī)制有關(guān)。3.構(gòu)建了 CiNAC1-GFP融合表達(dá)載體,并轉(zhuǎn)化野生型擬南芥的原生質(zhì)體中,通過(guò)激光共聚焦顯微鏡觀察其亞細(xì)胞定位,發(fā)現(xiàn)CiNAC1定位到細(xì)胞核中,這與它作為轉(zhuǎn)錄因子的功能是一致的。4.構(gòu)建了 CiNAC1-HA過(guò)表達(dá)載體并轉(zhuǎn)化野生型擬南芥,實(shí)時(shí)熒光定量RT-PCR檢測(cè)發(fā)現(xiàn)獲得的轉(zhuǎn)基因純合體株系中目的基因均有不同程度的表達(dá)。5.轉(zhuǎn)CiNAC1基因擬南芥株系側(cè)根數(shù)目顯著多于野生型;在NaC1和甘露醇處理下,CiNAC1過(guò)表達(dá)株系種子萌發(fā)率低于野生型;乙烯處理后,過(guò)表達(dá)CiNAC1的擬南芥提前衰老。
[Abstract]:After drought, high temperature, salt alkali, cold, insect pests and other stresses, cells will be damaged to varying degrees.Caragana intermedia has the characteristics of cold tolerance, drought resistance, saline-alkali tolerance and barren. NAC transcription factor family is one of the most important transcription factors family, which has good wind and sand prevention function and soil and water conservation function.It plays an important role in plant response to abiotic stress and growth and development.CiNAC1 gene was cloned from Caragana intermedia and its function was analyzed. The results are as follows: 1.The 1066 BP cDNA sequence of CiNAC1 gene of Caragana intermedia was cloned by PCR.Bioinformatics analysis showed that the open reading frame ORF of CiNAC1 gene was 921 BP, encoding 306 amino acids, the deduced molecular weight of the protein was 34.57 KD and the isoelectric point was 8.35. It was a kind of hydrophilic protein with conserved NAM domain at its N-terminal.There are 26 phosphorylation sites and 7 glycosylation sites.Real-time fluorescence quantitative RT-PCR analysis showed that the expression of CiNAC1 gene was induced by drought, high salt, dehydration and high pH, which suggested that the CiNAC1 gene might be related to the stress response mechanism of Caragana intermedia.The fusion expression vector of CiNAC1-GFP was constructed and transformed into the protoplasts of wild type Arabidopsis thaliana. The subcellular localization of CiNAC1 was observed by confocal laser microscopy. It was found that CiNAC1 was located in the nucleus, which was consistent with its function as a transcription factor.CiNAC1-HA overexpression vector was constructed and transformed into wild type Arabidopsis thaliana. Real-time fluorescence quantitative RT-PCR analysis showed that the target genes expressed in different degree in transgenic homozygous lines.The number of lateral roots of transgenic Arabidopsis thaliana lines with CiNAC1 gene was significantly more than that of wild type, the seed germination rate of NaC1 and mannitol overexpressed lines was lower than that of wild type lines, and that of Arabidopsis thaliana with CiNAC1 overexpressed after ethylene treatment was early senescence.
【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】：2017
【分類號(hào)】：Q943.2

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本文編號(hào)：1773079