转ClNAC9基因露地菊的耐盐碱性研究

发布时间：2018-06-24 02:21

本文选题：ClNAC9基因 + 露地菊　；参考：《东北林业大学》2016年硕士论文

【摘要】：露地菊‘纽9717’(Chrysanthemum×grandiflora 'niu9717')是地被菊‘纽扣粉’经卫星搭载后选育的一个新品种,植株低矮,花多而密,花色鲜艳,花期较长。NAC转录因子是高等植物中特有的转录因子。转入外源基因提高露地菊抗性对扩大其园林应用有重要意义。本文主要对转入甘菊(Chrysanthemum lavandulifolium)CINAC9基因的露地菊‘纽9717’的转录组及其对盐碱胁迫的耐受性进行研究,主要研究成果如下：1.通过高通量测序技术(High-throughput sequencing)对转基因露地菊进行转录组分析,转录组数据混合组装共获得91,182条Unigene序列。CINAC9-5株系与野生型露地菊相比显著差异表达的210个基因中,CINAC9-5株系对比野生型露地菊表达上调的有148个,表达下调的有62个。显著差异表达基因与各数据库进行比对后共注释到91个基因,其中有22个与盐碱胁迫应答相关,为研究转入外源基因CINAC9增强露地菊的耐盐碱性提供基因水平的依据。2.200mmol/L的NaCl盐溶液胁迫下,露地菊呈现出如下变化：形态上随着胁迫时间的延长植株逐渐出现受胁迫的症状,从叶片泛黄到植株地上部分全部萎蔫；组织相对含水量呈现下降趋势,相对电导率和MDA含量呈现不断上升的趋势,SOD、POD和CAT活性均呈现先升高后降低的趋势,但最高值出现的时间并不一致。形态和生理两方面的测定表明,3个转基因株系对盐胁迫的抗性明显高于野生型露地菊。3.胁迫20天时不同浓度盐溶液对露地菊的影响随着浓度的增加呈现出如下规律：露地菊的形态呈现出不同的变化,浓度越大,受迫害症状越严重；露地菊的相对含水量随着浓度的升高不断下降；相对电导率与MDA含量均呈上升趋势；抗氧化保护酶系统中SOD、 POD和CAT活性的变化不一,但各浓度胁迫中3个转基因株系的酶活性均显著高于野生型露地菊。试验表明,转基因株系比野生型露地菊更耐高浓度盐胁迫。4.100mmol/L碱溶液胁迫对露地菊的影响随着胁迫时间的延长呈现出如下规律：露地菊的形态呈现出不同的变化,但转基因株系的受害程度轻于野生型露地菊；植物过氧化产物MDA含量呈现上升的趋势；3个转基因株系的游离脯氨酸含量于胁迫15天后超过野生型植株,可溶性蛋白含量先小幅度上升后下降；过氧化保护酶SOD、POD和CAT活性均呈现先升高后降低的趋势,但最高值出现的时间不一致,但3个转基因株系的酶活性大多显著高于野生型露地菊。5.碱胁迫15天对露地菊的影响随着浓度的增加呈现出如下规律：相同浓度下转基因株系的形态受害程度远轻于野生型露地菊；随着浓度的升高MDA含量呈现出上升的趋势；游离脯氨酸含量在100mmol/L碱胁迫时迅速升高；转基因株系的可溶性蛋白含量均高于野生型露地菊；露地菊的抗氧化保护酶系统中SOD、POD和CAT酶活性呈现先升高后降低的趋势,在各浓度胁迫中3个转基因株系的酶活性均显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.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S682.11

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