【摘要】：為揭示組蛋白乙�；揎椩谑只ǹ谱魑锍檗钒l(fā)育表觀遺傳調(diào)控中的功能,以油青49和油青甜菜心80天的菜心為材料,通過(guò)PCR和RACE方法克隆了菜心BrcuHAC1基因c DNA和g DNA全長(zhǎng)序列,并進(jìn)行生物信息學(xué)分析,同時(shí)采用實(shí)時(shí)熒光定量PCR方法檢測(cè)目的基因的時(shí)空表達(dá)特性。結(jié)果表明,克隆獲得了BrcuHAC1基因全長(zhǎng)6 061 bp,包括CDS區(qū)5 067 bp,編碼1 688個(gè)氨基酸。對(duì)應(yīng)的g DNA全長(zhǎng)為7 376 bp,含有15個(gè)外顯子和14個(gè)內(nèi)含子,最長(zhǎng)的外顯子長(zhǎng)度為1 599 bp,內(nèi)含子的長(zhǎng)度范圍為50~210 bp。啟動(dòng)子序列中含有多個(gè)基本轉(zhuǎn)錄元件和順式作用元件,如光反應(yīng)元件、脫落酸響應(yīng)元件、參與厭氧誘導(dǎo)的增強(qiáng)子元件等。多序列比對(duì)結(jié)果表明,BrcuHAC1的氨基酸序列中含有高等植物HAC1基因的6個(gè)保守結(jié)構(gòu)域。系統(tǒng)發(fā)育分析結(jié)果顯示,菜心與油菜、甘藍(lán)、白菜處于同一分支,其親緣關(guān)系最近。時(shí)空特異性分析結(jié)果表明,BrcuHAC1在菜心的根、莖、花、葉中均有表達(dá),其中在花中表達(dá)量最高,葉次之,根最低;BrcuHAC1從苗期至完全抽薹開(kāi)花期的表達(dá)量呈現(xiàn)上升趨勢(shì),說(shuō)明BrcuHAC1在菜薹花發(fā)育過(guò)程中可能起到一定的調(diào)控作用。本研究結(jié)果為揭示組蛋白乙酰轉(zhuǎn)移酶調(diào)控菜心抽薹分子機(jī)制提供了一定的理論依據(jù)。
[Abstract]:In order to reveal the function of histone acetylation modification in epigenetic regulation of bolting and development of cruciferous crops, the full-length sequences of c DNA and g DNA of BrcuHAC1 gene were cloned by PCR and RACE. Bioinformatics analysis and real-time fluorescence quantitative PCR were used to detect the temporal and spatial expression characteristics of the target gene. The results showed that the full length of BrcuHAC1 gene was 6 061 BP, including CDS region 5 067 BP, encoding 1 688 amino acids. The total length of g DNA is 7 376 BP, which contains 15 exons and 14 introns. The longest exon length is 1 599 BP, and the length range of intron is 50 ~ 210bp. The promoter sequence contains a number of basic transcription elements and cis-acting elements, such as photoreactive elements, abscisic acid response elements, anaerobic-induced enhancers, and so on. The results of multiple sequence alignment showed that the amino acid sequence of BrcuHAC1 contained six conserved domains of HAC1 gene of higher plants. Phylogenetic analysis showed that Brassica rapeseed, Brassica napus and Brassica campestris were in the same branch and had the closest phylogenetic relationship. The results of spatio-temporal specificity analysis showed that BrcuHAC1 was expressed in root, stem, flower and leaf of Chinese cabbage, among which the highest expression was in flower, followed by leaf, and the lowest BrcuHAC1 in root showed an increasing trend from seedling stage to full bolting stage. The results suggest that BrcuHAC1 may play a regulating role in the development of cauliflower. The results provide a theoretical basis for revealing the molecular mechanism of histone acetyltransferase regulating bolting of Chinese cabbage.
【作者單位】： 江西農(nóng)業(yè)大學(xué)農(nóng)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(31360484) 教育部高等學(xué)校博士學(xué)科點(diǎn)專項(xiàng)科研基金(20133603120006) 江西省自然科學(xué)基金項(xiàng)目(2013BAB204009)
【分類號(hào)】：Q943.2;S634.5
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本文編號(hào)：2164053