本文關(guān)鍵詞： 巨桉 NAC基因 非生物逆境 基因表達(dá)　出處：《林業(yè)科學(xué)》2017年10期 　論文類型：期刊論文

【摘要】：【目的】NAC(NAM,ATAF和CUC2蛋白)是植物中一類特異轉(zhuǎn)錄因子,廣泛參與植物生長、發(fā)育、激素信號(hào)轉(zhuǎn)導(dǎo)和逆境響應(yīng)過程。本文通過對(duì)巨桉EgrNAC1(Eucgr.I00058)編碼蛋白結(jié)構(gòu)、蛋白亞細(xì)胞定位特點(diǎn),以及低溫、干旱和高鹽等非生物逆境條件下的基因表達(dá)分析,研究EgrNAC1基因與非生物逆境響應(yīng)的關(guān)系,為其功能的深入研究提供基礎(chǔ)�！痉椒ā渴紫葟木掼窕蚪M數(shù)據(jù)庫下載EgrNAC1基因、啟動(dòng)子和編碼蛋白序列,利用SMART、MatInspector和MEGA等軟件對(duì)EgrNAC1蛋白結(jié)構(gòu)特征、進(jìn)化分類及啟動(dòng)子上的順式作用元件進(jìn)行分析;并以pCAMBIA1300為基礎(chǔ)載體,用酶切法構(gòu)建EgrNAC1∷sGFP載體,采用基因槍轟擊洋蔥表皮方法,對(duì)EgrNAC1蛋白表達(dá)的亞細(xì)胞定位特點(diǎn)進(jìn)行研究。然后,以4℃不同時(shí)間處理的數(shù)字表達(dá)譜數(shù)據(jù)為基礎(chǔ),利用WGCNA軟件進(jìn)行基因共表達(dá)分析,了解低溫下與EgrNAC1高度相關(guān)的共表達(dá)基因特征。為進(jìn)一步了解EgrNAC1在不同非生物逆境處理下的表達(dá)特征,同樣利用3個(gè)月巨桉無性系幼苗進(jìn)行不同低溫(-8,-4,0,4,8℃)、4℃不同時(shí)間(2,6,12,24,48 h)、高溫(42℃)、高鹽、干旱、脫落酸(ABA)和茉莉酸甲酯(MeJA)的不同處理,用實(shí)時(shí)熒光定量RT-PCR方法對(duì)這些處理下EgrNAC1的表達(dá)情況進(jìn)行分析�！窘Y(jié)果】EgrNAC1中NAC結(jié)構(gòu)域包含A,B,C,D,E 5個(gè)典型亞結(jié)構(gòu)域,其中含2個(gè)α螺旋5個(gè)β折疊片結(jié)構(gòu),有核定位序列,無跨膜域。進(jìn)化樹構(gòu)建結(jié)果表明,EgrNAC1屬于NAC家族的Ⅰ類亞家族ATAF子組,逆境類NAC分類中屬于SNAC-A亞類。EgrNAC1啟動(dòng)子序列上含有ABRE、MBS、MCS和DREB等大量與逆境脅迫相關(guān)的順式作用元件。亞細(xì)胞定位結(jié)果表明EgrNAC1在核中表達(dá)。4℃不同處理時(shí)間(0,2,6,12,24,48 h)下與EgrNAC1共表達(dá)相關(guān)系數(shù)最高的20個(gè)基因中,大多數(shù)基因參與逆境脅迫響應(yīng)相關(guān)的調(diào)控過程。不同時(shí)間處理下,葉片中EgrNAC1誘導(dǎo)水平隨處理時(shí)間延長不斷提高;不同低溫下,4℃和8℃處理中EgrNAC1的誘導(dǎo)水平相對(duì)較高;干旱處理1天后基因表達(dá)即受到誘導(dǎo),然后又有所下降;鹽(200 mmol·L~(-1)NaCl)脅迫48 h,才能誘導(dǎo)EgrNAC1表達(dá);100μmol·L~(-1)ABA和100μmol·L~(-1)MeJA均能誘導(dǎo)葉片中EgrNAC1基因表達(dá),MeJA誘導(dǎo)速度更快,處理2 h后基因表達(dá)即明顯提高,而ABA的誘導(dǎo)效應(yīng)則需要24 h�！窘Y(jié)論】EgrNAC1是參與巨桉逆境脅迫響應(yīng)的1個(gè)NAC類轉(zhuǎn)錄因子基因,其不僅參與低溫、干旱和高鹽的非生物逆境脅迫響應(yīng),還可能與ABA、MeJA信號(hào)轉(zhuǎn)導(dǎo)有交叉互作效應(yīng)。
[Abstract]:[objective] NACU NAM ATAF and CUC2 proteins are a kind of specific transcription factors in plants, which are widely involved in plant growth and development. Hormone signal transduction and stress response. EgrNAC1Eucgr.I00058 (EgrNAC1Eucgr.I00058) encodes protein structure, subcellular localization, and hypothermia. The relationship between EgrNAC1 gene and abiotic stress response was studied by analyzing the gene expression under abiotic stress conditions such as drought and high salt. [methods] EgrNAC1 gene, promoter and coding protein sequence were downloaded from Eucalyptus grandis genome database and SMART was used. MatInspector and MEGA software were used to analyze the structural characteristics, evolutionary classification and cis-acting elements of EgrNAC1 protein. The EgrNAC1: sGFP vector was constructed by enzyme digestion with pCAMBIA1300 as the basic vector, and the onion epidermis was bombarded with gene gun. The subcellular localization characteristics of EgrNAC1 protein expression were studied. Then, based on the data of digital expression profile at 4 鈩，

本文編號(hào)：1458170