擬南芥微絲結(jié)合蛋白ADF1超表達(dá)構(gòu)建及其在鹽脅迫下的作用
發(fā)布時(shí)間：2018-08-31 19:53
【摘要】：土壤鹽堿化是影響農(nóng)作物產(chǎn)量的一個(gè)非常主要的因素。鹽分過高可以使植物體內(nèi)的離子失衡,造成滲透脅迫等一系列非生物脅迫,影響植物的正常生長。因此我們對植物耐鹽機(jī)理的研究不僅能夠豐富植物響應(yīng)逆境過程的理論知識(shí),其研究的結(jié)果還可以用于農(nóng)業(yè)生產(chǎn)實(shí)踐,解決培育高耐鹽性的農(nóng)作物的問題。本實(shí)驗(yàn)室前期研究發(fā)現(xiàn),微絲結(jié)合蛋白基因AtADF1可以被鹽脅迫誘導(dǎo),AtADF1突變體造成了植物對鹽敏感,阻礙了鹽誘導(dǎo)的前期微絲解聚過程。研究結(jié)果表明,微絲結(jié)合蛋白基因AtADF1的缺失影響擬南芥響應(yīng)鹽脅迫的過程。本實(shí)驗(yàn)首先通過克隆、構(gòu)建、鑒定的方法獲得了 3個(gè)擬南芥超表達(dá)ADF1(AtADF1-OEs)純合株系,然后以擬南芥野生型(WT)和2個(gè)純合株系的AtADF1-OEs為實(shí)驗(yàn)材料,觀察擬南芥野生型(WT)和AtADF1-OEs純合植株在鹽脅迫下的萌發(fā)率,生長情況和微絲的動(dòng)態(tài)變化,測定鹽脅迫下擬南芥AtADF1-OEs純合植株中鹽脅迫響應(yīng)基因SOS1、SOS2和SOS3的相對表達(dá)量,從而揭示超表達(dá)擬南芥微絲結(jié)合蛋白基因ADF1在鹽脅迫下的作用機(jī)制。主要結(jié)果如下:(1)通過克隆AtADF1基因,構(gòu)建AtADF1超表達(dá)載體,從而得到了AtADF1的超表達(dá)植株。(2)對AtADF1的超表達(dá)植株進(jìn)行篩選和鑒定,總共獲得了 3個(gè)不同表達(dá)量的純合植株 AtADF1-OEs。(3)在正常條件下,擬南芥野生型(WT)和AtADF1-OEs植株的萌發(fā)率基本相同。在鹽脅迫下(NaCl處理),AtADF1-OEs植株的萌發(fā)率要高于WT。(4)在正常條件下,擬南芥野生型(WT)和AtADF1-OEs純合植株的生長情況基本一致。在鹽脅迫下(NaCl處理),AtADF1-OEs純合植株的生長趨勢要強(qiáng)于WT植株,主要表現(xiàn)為側(cè)根數(shù)、葉面積和根長均相對變大。(5)在正常條件下,擬南芥野生型(WT)和AtADF1-OE植株的微絲都是聚合的,沒有發(fā)生解聚。在鹽脅迫下(NaCl處理),與WT相比,AtADF1-OE植株增強(qiáng)了微絲的解聚。(6)在正常條件下,AtADF1-OEs純合植株中SOS1、SOS2和SOS3基因表達(dá)量與WT植株相比基本相同。在鹽脅迫下(NaCl處理),WT植株中SOS1、SOS2和SOS3基因表達(dá)量都是升高的;AtADF1-OEs純合植株中SOS2和SOS3基因的表達(dá)量都高于WT植株。綜上所述,AtADF1基因超表達(dá)后,有利于鹽脅迫下植物的萌發(fā)和生長過程,增強(qiáng)了鹽誘導(dǎo)的微絲解聚過程,提高了鹽誘導(dǎo)的SOS2和SOS3基因的表達(dá),說明AtADF1基因超表達(dá)提高了擬南芥耐鹽的能力。
[Abstract]:Soil salinization is a very important factor affecting crop yield. Excessive salinity can cause ion imbalance in plants, resulting in a series of abiotic stresses, such as osmotic stress, which affect the normal growth of plants. Therefore, our research on the mechanism of plant salt tolerance can not only enrich the theoretical knowledge of plant response to stress, but also can be used in agricultural production to solve the problem of cultivating crops with high salt tolerance. Our previous study found that the AtADF1 gene of microfilament binding protein could be induced by salt stress to induce AtADF1 mutant to make plants sensitive to salt and hinder the process of microfilament depolymerization induced by salt. The results showed that the deletion of microfilament binding protein (AtADF1) gene affected the response of Arabidopsis thaliana to salt stress. In this experiment, three Arabidopsis superexpressed ADF1 (AtADF1-OEs) homozygous lines were obtained by cloning, construction and identification, and then the wild-type (WT) and AtADF1-OEs of two homozygous strains were used as experimental materials. The germination rate, growth and microfilament dynamics of wild-type Arabidopsis (WT) and AtADF1-OEs homozygous plants under salt stress were observed. The relative expression of SOS1,SOS2 and SOS3 genes in Arabidopsis AtADF1-OEs homozygous plants were measured. The mechanism of overexpression of microfilament binding protein (ADF1) gene in Arabidopsis thaliana under salt stress was revealed. The main results are as follows: (1) by cloning the AtADF1 gene and constructing the AtADF1 overexpression vector, the superexpression plants of AtADF1 were obtained. (2) the superexpression plants of AtADF1 were screened and identified. Three homozygous plants AtADF1-OEs. (3) with different expression levels were obtained. Under normal conditions, the germination rates of wild-type (WT) and AtADF1-OEs plants in Arabidopsis thaliana were almost the same. The germination rate of AtADF1-OEs plants under salt stress (NaCl treatment) was higher than that of WT. (4) under normal conditions, the growth of wild type (WT) and AtADF1-OEs homozygous plants of Arabidopsis thaliana was basically the same. Under salt stress (NaCl treatment), the growth trend of AtADF1-OEs homozygous plants was stronger than that of WT plants, mainly showing that the number of lateral roots, leaf area and root length were relatively larger. (5) under normal conditions, the microfilaments of Arabidopsis wild-type (WT) and AtADF1-OE plants were polymerized. There was no depolymerization. Under salt stress (NaCl treatment), AtADF1-OE enhanced the depolymerization of microfilaments compared with WT. (6) under normal conditions, the expression of SOS1,SOS2 and SOS3 genes in AtADF1-OEs homozygous plants was almost the same as that in WT plants. Under salt stress (NaCl treatment), the expression of SOS1,SOS2 and SOS3 genes in the homozygous plants of AtADF1-OEs was higher than that in WT plants. In conclusion, the overexpression of AtADF1 gene was beneficial to the germination and growth of plants under salt stress, enhanced the process of microfilament depolymerization induced by salt, and enhanced the expression of SOS2 and SOS3 genes induced by salt. The results showed that the overexpression of AtADF1 gene enhanced the salt tolerance of Arabidopsis thaliana.
【學(xué)位授予單位】：沈陽農(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;Q945.78
本文編號(hào)：2216001
[Abstract]:Soil salinization is a very important factor affecting crop yield. Excessive salinity can cause ion imbalance in plants, resulting in a series of abiotic stresses, such as osmotic stress, which affect the normal growth of plants. Therefore, our research on the mechanism of plant salt tolerance can not only enrich the theoretical knowledge of plant response to stress, but also can be used in agricultural production to solve the problem of cultivating crops with high salt tolerance. Our previous study found that the AtADF1 gene of microfilament binding protein could be induced by salt stress to induce AtADF1 mutant to make plants sensitive to salt and hinder the process of microfilament depolymerization induced by salt. The results showed that the deletion of microfilament binding protein (AtADF1) gene affected the response of Arabidopsis thaliana to salt stress. In this experiment, three Arabidopsis superexpressed ADF1 (AtADF1-OEs) homozygous lines were obtained by cloning, construction and identification, and then the wild-type (WT) and AtADF1-OEs of two homozygous strains were used as experimental materials. The germination rate, growth and microfilament dynamics of wild-type Arabidopsis (WT) and AtADF1-OEs homozygous plants under salt stress were observed. The relative expression of SOS1,SOS2 and SOS3 genes in Arabidopsis AtADF1-OEs homozygous plants were measured. The mechanism of overexpression of microfilament binding protein (ADF1) gene in Arabidopsis thaliana under salt stress was revealed. The main results are as follows: (1) by cloning the AtADF1 gene and constructing the AtADF1 overexpression vector, the superexpression plants of AtADF1 were obtained. (2) the superexpression plants of AtADF1 were screened and identified. Three homozygous plants AtADF1-OEs. (3) with different expression levels were obtained. Under normal conditions, the germination rates of wild-type (WT) and AtADF1-OEs plants in Arabidopsis thaliana were almost the same. The germination rate of AtADF1-OEs plants under salt stress (NaCl treatment) was higher than that of WT. (4) under normal conditions, the growth of wild type (WT) and AtADF1-OEs homozygous plants of Arabidopsis thaliana was basically the same. Under salt stress (NaCl treatment), the growth trend of AtADF1-OEs homozygous plants was stronger than that of WT plants, mainly showing that the number of lateral roots, leaf area and root length were relatively larger. (5) under normal conditions, the microfilaments of Arabidopsis wild-type (WT) and AtADF1-OE plants were polymerized. There was no depolymerization. Under salt stress (NaCl treatment), AtADF1-OE enhanced the depolymerization of microfilaments compared with WT. (6) under normal conditions, the expression of SOS1,SOS2 and SOS3 genes in AtADF1-OEs homozygous plants was almost the same as that in WT plants. Under salt stress (NaCl treatment), the expression of SOS1,SOS2 and SOS3 genes in the homozygous plants of AtADF1-OEs was higher than that in WT plants. In conclusion, the overexpression of AtADF1 gene was beneficial to the germination and growth of plants under salt stress, enhanced the process of microfilament depolymerization induced by salt, and enhanced the expression of SOS2 and SOS3 genes induced by salt. The results showed that the overexpression of AtADF1 gene enhanced the salt tolerance of Arabidopsis thaliana.
【學(xué)位授予單位】：沈陽農(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;Q945.78
【相似文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 劉宇昌;擬南芥微絲結(jié)合蛋白ADF1超表達(dá)構(gòu)建及其在鹽脅迫下的作用[D];沈陽農(nóng)業(yè)大學(xué);2017年
，本文編號(hào)：2216001
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