本文選題：小麥 + b�。� 參考：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2016年02期

【摘要】：堿性亮氨酸拉鏈(basic leucine zipper,b ZIP)是植物中廣泛存在的一類轉(zhuǎn)錄因子,參與多種脅迫響應(yīng)與生長發(fā)育過程。本研究從小麥(Triticum aestivum)中克隆到一個(gè)熱脅迫誘導(dǎo)的b ZIP家族轉(zhuǎn)錄因子基因Tab ZIP28(Gen Bank登錄號(hào):KT753298.1),ORF長度為1 713 bp,編碼570個(gè)氨基酸。生物信息學(xué)分析結(jié)果表明,Tab ZIP28與擬南芥b ZIP家族轉(zhuǎn)錄因子中B亞組的3個(gè)基因Atb ZIP17、Atb ZIP28和Atb ZIP49歸為一類。氨基酸序列比對(duì)結(jié)果表明,該蛋白具有b ZIP和跨膜結(jié)構(gòu)域(transmembrane domain,TMD)兩個(gè)保守結(jié)構(gòu)域以及規(guī)范的位點(diǎn)1蛋白酶(site 1 protease,S1P)剪切位點(diǎn)。對(duì)該基因起始密碼子ATG上游1 699bp的序列進(jìn)行順式作用元件分析,發(fā)現(xiàn)該基因的啟動(dòng)子區(qū)域包含眾多激素和逆境脅迫響應(yīng)元件。通過q RT-PCR對(duì)該基因在逆境脅迫下的表達(dá)模式進(jìn)行分析,結(jié)果表明,Tab ZIP28在熱脅迫處理1 h即上調(diào)表達(dá)且達(dá)到最大值;用20%PEG 6000模擬干旱環(huán)境處理小麥幼苗后,Tab ZIP28在處理6 h達(dá)到最大值,并在12 h時(shí)急劇下降;對(duì)5 mmol/L H2O2處理響應(yīng)比較緩慢,在處理12 h才上調(diào)表達(dá);該基因不受到二硫蘇糖醇(dithiothreitol,DTT)處理的誘導(dǎo)表達(dá)。在擬南芥(Arabidopsis thaliana)中過量表達(dá)Tab ZIP28基因,轉(zhuǎn)基因株系在高溫脅迫后的成活率和種子發(fā)芽率較野生型明顯提高,說明該基因可能對(duì)植物的耐熱性有貢獻(xiàn),可以作為耐熱性育種的候選基因。
[Abstract]:Alkaline leucine zipper basic leucine zipperb (ZIP) is a widespread transcription factor in plants, which is involved in many stress response and growth and development processes. In this study, a heat stress induced b ZIP family transcription factor gene, Tab ZIP28(Gen Bank accession number, was cloned from Triticum aestivum. The length of Tab ZIP28(Gen Bank was 1 713 BP, encoding 570 amino acids. Bioinformatics analysis showed that Tab ZIP28 and three genes of B subgroup Atb ZIP17Ab ZIP28 and Atb ZIP49 in Arabidopsis thaliana B ZIP family were classified into one group. The results of amino acid sequence alignment showed that the protein had two conserved domains, b ZIP and transmembrane domain (TMD), as well as the normal site 1 protease S1P. The sequence of 1 699bp upstream of the initiation codon of the gene was analyzed by cis-acting element analysis. It was found that the promoter region of the gene contained many hormones and stress response elements. Q RT-PCR was used to analyze the expression pattern of the gene under stress. The results showed that Tab ZIP28 was up-regulated at 1 h of heat stress and reached its maximum value. 20%PEG 6000 was used to simulate the drought environment of wheat seedlings, and Tab ZIP28 reached its maximum value at 6 h and decreased sharply at 12 h, and the response to 5 mmol/L H2O2 treatment was slow, and the expression of Tab ZIP28 was up-regulated at 12 h after treatment. The gene was not induced by dithiothreitol DTT treatment. The overexpression of Tab ZIP28 gene in Arabidopsis thaliana of Arabidopsis showed that the survival rate and seed germination rate of transgenic lines after high temperature stress were significantly higher than that of wild type, which suggested that the gene might contribute to the heat tolerance of plants. It can be used as a candidate gene for heat resistance breeding.
【作者單位】： 中國農(nóng)業(yè)大學(xué)農(nóng)學(xué)院/植物遺傳育種學(xué)系/農(nóng)業(yè)生物技術(shù)國家重點(diǎn)實(shí)驗(yàn)室/北京市作物遺傳改良重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金(No.31171549) 轉(zhuǎn)基因?qū)ｍ?xiàng)(No.2011ZX08002-002)
【分類號(hào)】：S512.1

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