本文選題：ClNAC9基因 + 露地菊��； 參考：《東北林業(yè)大學(xué)》2016年碩士論文

【摘要】：露地菊‘紐9717’(Chrysanthemum×grandiflora 'niu9717')是地被菊‘紐扣粉’經(jīng)衛(wèi)星搭載后選育的一個(gè)新品種,植株低矮,花多而密,花色鮮艷,花期較長(zhǎng)。NAC轉(zhuǎn)錄因子是高等植物中特有的轉(zhuǎn)錄因子。轉(zhuǎn)入外源基因提高露地菊抗性對(duì)擴(kuò)大其園林應(yīng)用有重要意義。本文主要對(duì)轉(zhuǎn)入甘菊(Chrysanthemum lavandulifolium)CINAC9基因的露地菊‘紐9717’的轉(zhuǎn)錄組及其對(duì)鹽堿脅迫的耐受性進(jìn)行研究,主要研究成果如下：1.通過(guò)高通量測(cè)序技術(shù)(High-throughput sequencing)對(duì)轉(zhuǎn)基因露地菊進(jìn)行轉(zhuǎn)錄組分析,轉(zhuǎn)錄組數(shù)據(jù)混合組裝共獲得91,182條Unigene序列。CINAC9-5株系與野生型露地菊相比顯著差異表達(dá)的210個(gè)基因中,CINAC9-5株系對(duì)比野生型露地菊表達(dá)上調(diào)的有148個(gè),表達(dá)下調(diào)的有62個(gè)。顯著差異表達(dá)基因與各數(shù)據(jù)庫(kù)進(jìn)行比對(duì)后共注釋到91個(gè)基因,其中有22個(gè)與鹽堿脅迫應(yīng)答相關(guān),為研究轉(zhuǎn)入外源基因CINAC9增強(qiáng)露地菊的耐鹽堿性提供基因水平的依據(jù)。2.200mmol/L的NaCl鹽溶液脅迫下,露地菊呈現(xiàn)出如下變化：形態(tài)上隨著脅迫時(shí)間的延長(zhǎng)植株逐漸出現(xiàn)受脅迫的癥狀,從葉片泛黃到植株地上部分全部萎蔫；組織相對(duì)含水量呈現(xiàn)下降趨勢(shì),相對(duì)電導(dǎo)率和MDA含量呈現(xiàn)不斷上升的趨勢(shì),SOD、POD和CAT活性均呈現(xiàn)先升高后降低的趨勢(shì),但最高值出現(xiàn)的時(shí)間并不一致。形態(tài)和生理兩方面的測(cè)定表明,3個(gè)轉(zhuǎn)基因株系對(duì)鹽脅迫的抗性明顯高于野生型露地菊。3.脅迫20天時(shí)不同濃度鹽溶液對(duì)露地菊的影響隨著濃度的增加呈現(xiàn)出如下規(guī)律：露地菊的形態(tài)呈現(xiàn)出不同的變化,濃度越大,受迫害癥狀越嚴(yán)重；露地菊的相對(duì)含水量隨著濃度的升高不斷下降；相對(duì)電導(dǎo)率與MDA含量均呈上升趨勢(shì)；抗氧化保護(hù)酶系統(tǒng)中SOD、 POD和CAT活性的變化不一,但各濃度脅迫中3個(gè)轉(zhuǎn)基因株系的酶活性均顯著高于野生型露地菊。試驗(yàn)表明,轉(zhuǎn)基因株系比野生型露地菊更耐高濃度鹽脅迫。4.100mmol/L堿溶液脅迫對(duì)露地菊的影響隨著脅迫時(shí)間的延長(zhǎng)呈現(xiàn)出如下規(guī)律：露地菊的形態(tài)呈現(xiàn)出不同的變化,但轉(zhuǎn)基因株系的受害程度輕于野生型露地菊；植物過(guò)氧化產(chǎn)物MDA含量呈現(xiàn)上升的趨勢(shì)；3個(gè)轉(zhuǎn)基因株系的游離脯氨酸含量于脅迫15天后超過(guò)野生型植株,可溶性蛋白含量先小幅度上升后下降；過(guò)氧化保護(hù)酶SOD、POD和CAT活性均呈現(xiàn)先升高后降低的趨勢(shì),但最高值出現(xiàn)的時(shí)間不一致,但3個(gè)轉(zhuǎn)基因株系的酶活性大多顯著高于野生型露地菊。5.堿脅迫15天對(duì)露地菊的影響隨著濃度的增加呈現(xiàn)出如下規(guī)律：相同濃度下轉(zhuǎn)基因株系的形態(tài)受害程度遠(yuǎn)輕于野生型露地菊；隨著濃度的升高M(jìn)DA含量呈現(xiàn)出上升的趨勢(shì)；游離脯氨酸含量在100mmol/L堿脅迫時(shí)迅速升高；轉(zhuǎn)基因株系的可溶性蛋白含量均高于野生型露地菊；露地菊的抗氧化保護(hù)酶系統(tǒng)中SOD、POD和CAT酶活性呈現(xiàn)先升高后降低的趨勢(shì),在各濃度脅迫中3個(gè)轉(zhuǎn)基因株系的酶活性均顯著高于野生型露地菊。
[Abstract]:'Chrysanthemum 脳 grandiflora Niu 9717' (Chrysanthemum 脳 grandiflora Niu 9717') is a new variety selected by satellite. Its plant is low, flowers are many and dense, its color is bright, and its flowering period is longer. NAC transcription factor is a unique transcription factor in higher plants. It is very important to increase the resistance of chrysanthemum to foreign genes in order to expand its garden application. The transcriptome of chrysanthemum lavandulifolium CINAC9 gene and its tolerance to saline-alkali stress were studied in this paper. The main results are as follows: 1. Transcriptome analysis of transgenic open-field chrysanthemum was carried out by high-throughput sequencing. A total of 91182 Unigene sequences. CINAC9-5 strains were significantly differentially expressed in comparison with wild type open chrysanthemum. 148 of them were up-regulated and 62 were down-regulated. Significant differentially expressed genes were annotated to 91 genes compared with each database, 22 of which were related to saline-alkali stress response. In order to study the gene level of the exogenous gene CINAC9 to enhance the saline-alkali tolerance of the chrysanthemum, under NaCl salt solution stress of 2.200 mmol / L, the plant showed the following changes: morphologically, with the extension of the stress time, the plant gradually appeared the symptom of stress. From yellowing leaves to wilting above ground parts, the relative water content of tissues decreased, and the relative electrical conductivity and MDA content showed an increasing trend. The activities of POD and cat increased first and then decreased. But the maximum appears at different times. The results of morphological and physiological analysis showed that the resistance of the three transgenic lines to salt stress was significantly higher than that of wild type open chrysanthemum. 3. After 20 days of stress, the effects of different concentrations of salt solution on the chrysanthemum were as follows: the morphology of the chrysanthemum showed different changes, the greater the concentration, the more serious the persecuted symptoms; The relative water content of chrysanthemum decreased with the increase of concentration, the relative conductivity and MDA content increased, and the activities of SOD, POD and cat in antioxidant protective enzyme system were different. However, the enzyme activity of three transgenic lines was significantly higher than that of wild type open chrysanthemum under different concentration stress. The results showed that transgenic lines were more tolerant to high concentration of salt stress than wild type open chrysanthemum. The effect of alkali solution stress of 4.100 mmol / L on chrysanthemum showed the following regularity: the morphology of open field chrysanthemum showed different changes with the prolongation of stress time. However, the damage of transgenic lines was lighter than that of wild type open chrysanthemum, the MDA content of plant peroxidation products showed an increasing trend, and the free proline content of three transgenic lines was higher than that of wild type plants after 15 days of stress. The content of soluble protein increased at first and then decreased, and the activities of POD and cat increased first and then decreased, but the highest value appeared at different time. However, the enzyme activity of the three transgenic lines was significantly higher than that of wild type chrysanthemum. The effect of alkali stress on open field chrysanthemum in 15 days showed the following regularity with the increase of concentration: the morphological damage degree of transgenic lines at the same concentration was much lighter than that of wild type open chrysanthemum, and the MDA content increased with the increase of concentration. The content of free proline increased rapidly under 100 mmol / L alkali stress, the soluble protein content of transgenic lines was higher than that of wild type open chrysanthemum, and the activities of SOD POD and cat in the antioxidant and protective enzyme system were increased first and then decreased. The enzyme activity of three transgenic lines was significantly higher than that of wild type open field chrysanthemum under different concentration stress.
【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)】：S682.11

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1 蔡英杰;轉(zhuǎn)ClNAC9基因露地菊的耐鹽堿性研究[D];東北林業(yè)大學(xué);2016年

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本文編號(hào)：2059553