本文選題：檸條 + LEA基因　； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：干旱、寒冷、鹽堿等是內(nèi)蒙地區(qū)常見的逆境因子,給當(dāng)?shù)刈魑锏纳L(zhǎng)、發(fā)育及產(chǎn)量造成嚴(yán)重影響,使農(nóng)業(yè)經(jīng)濟(jì)遭受損失,因此關(guān)于植物抗逆境脅迫的相關(guān)研究便顯得尤為重要。胚胎發(fā)育晚期豐富蛋白LEA作為植物中廣泛存在的一類逆境脅迫響應(yīng)蛋白受到了很多研究者的關(guān)注。本研究從檸條錦雞兒(Caragana korshinskii Kom.)中克隆得到兩個(gè)LEA蛋白基因CkLEA4-1和CkLEA6-1,并圍繞其脅迫相關(guān)的生物學(xué)功能展開研究,所得結(jié)果如下:1.CkLEA4-1基因的開放閱讀框長(zhǎng)為1119bp,編碼373個(gè)氨基酸;CkLEA6-1基因的開放閱讀框長(zhǎng)為255bp,編碼85個(gè)氨基酸。經(jīng)DNAMAN軟件親疏水性分析兩個(gè)LEA蛋白均為親水蛋白。序列比對(duì)與系統(tǒng)進(jìn)化分析表明,CkLEA4-1屬于LEA_4蛋白一組,但無(wú)保守結(jié)構(gòu)域,因此命名為CkLEA4-1;CkLEA6-1具有LEA_6保守結(jié)構(gòu)域因此命名該基因?yàn)镃kLEA6-1。2.實(shí)時(shí)熒光定量PCR檢測(cè)發(fā)現(xiàn)CkLEA4-1的表達(dá)受干旱、脫水、高鹽脅迫的誘導(dǎo),而CkLEA6-1的表達(dá)則主要受冷脅迫誘導(dǎo)。3.甘露醇處理下,CkLEA4-1基因過表達(dá)擬南芥種子萌發(fā)率與野生型無(wú)明顯差異,但子葉綠化率卻高于野生型。甘露醇和NaCl脅迫處理下,CkLEA4-1基因過表達(dá)擬南芥幼苗的根長(zhǎng)明顯比野生型長(zhǎng),且400mM甘露醇處理下過表達(dá)株系幼苗比野生型的略大、鮮重增加。干旱脅迫處理下,生長(zhǎng)4周的CkLEA4-1基因過表達(dá)擬南芥對(duì)干旱的耐受能力增強(qiáng),MDA含量也明顯低于野生型。4.冷脅迫處理下CkLEA6-1基因過表達(dá)擬南芥幼苗的根長(zhǎng)明顯比野生型長(zhǎng),且幼苗鮮重略高于野生型。此外在低溫脅迫下,CkLEA6-1基因過表達(dá)擬南芥質(zhì)膜的相對(duì)電導(dǎo)率在常溫、4℃時(shí)無(wú)明顯差別,在-20℃時(shí)過表達(dá)株系較野生型略低,過表達(dá)株系的MDA含量也低于野生型。
[Abstract]:Drought, cold, salt and alkali are common stress factors in Inner Mongolia, which have a serious impact on the growth, development and yield of local crops and cause losses to agricultural economy. Therefore, it is particularly important to study the resistance of plants to stress.LEA, a rich protein in late embryonic development, has attracted much attention from many researchers as a kind of stress response proteins widely existed in plants.In this study, Caragana korshinskii Kom.from Caragana korshinskiiTwo LEA protein genes CkLEA4-1 and CkLEA6-1 were cloned, and their biological functions related to stress were studied.The results are as follows: 1. The open reading frame length of CkLEA4-1 gene is 1119 BP, and the open reading frame length of 373 amino acid gene CkLEA6-1 gene is 255 BP, encoding 85 amino acids.The hydrophobicity of two LEA proteins was analyzed by DNAMAN software.Sequence alignment and phylogenetic analysis showed that CkLEA4-1 belongs to a group of LEA_4 proteins, but has no conserved domain. Therefore, CkLEA4-1CkLEA6-1 has LEA_6 conserved domain, so it is named as CkLEA6-1.2.Real-time fluorescence quantitative PCR analysis showed that the expression of CkLEA4-1 was induced by drought, dehydration and high salt stress, while the expression of CkLEA6-1 was mainly induced by cold stress.Under mannitol treatment, the germination rate of Arabidopsis thaliana seeds was not significantly different from that of wild type, but the greening rate of cotyledon was higher than that of wild type.Under mannitol and NaCl stress, the root length of overexpressed Arabidopsis thaliana seedlings was significantly longer than that of wild type seedlings, and the overexpressed strain seedlings under 400mM mannitol treatment were slightly larger than that of wild type seedlings, and the fresh weight of overexpressed Arabidopsis thaliana seedlings increased.Under drought stress, the overexpression of CkLEA4-1 gene in Arabidopsis thaliana was significantly lower than that in wild type.Under cold stress, the root length of Arabidopsis thaliana overexpressed by CkLEA6-1 gene was longer than that of wild type, and the fresh weight of seedlings was slightly higher than that of wild type.In addition, under low temperature stress, the relative conductivity of plasma membrane of Arabidopsis thaliana overexpressed CkLEA6-1 gene was not significantly different at room temperature (4 鈩，

本文編號(hào)：1764884