發(fā)布時間：2018-05-04 20:29

  本文選題：甘藍(lán)型油菜 + 角質(zhì)層蠟質(zhì)��； 參考：《作物學(xué)報》2017年05期

【摘要】：在植物蠟質(zhì)合成途徑中,中鏈烷烴羥化酶(mid-chain alkane hydroxylase,MAH)催化烷烴羥基化形成二級醇,進(jìn)一步氧化為酮。本研究以擬南芥P450依賴性酶CYP96A15/MAH1基因為探針,采用電子克隆與RT-PCR技術(shù),獲得2個甘藍(lán)型油菜MAH1的全長編碼區(qū)序列,分別命名為BnMAH1-1和BnMAH1-2(Gen Bank登錄號分別為KT795344和KT795345)。二者ORF長度均為1491 bp,無內(nèi)含子,核苷酸與氨基酸序列分別有92.4%與90.9%的一致性。根據(jù)編碼區(qū)預(yù)測的BnMAH1-1和BnMAH1-2前體蛋白均為包含496個氨基酸殘基的多肽鏈,具有典型的P450蛋白家族保守結(jié)構(gòu)P415x R417x、K螺旋(E359xx R362)、C末端的血紅素結(jié)合域(F436xx Gx Rx Cx G445)以及氧結(jié)合帶保守區(qū)域(A/G)G309x(D/E)T312(T/S)。NCBI Blast N、氨基酸序列多重比對與系統(tǒng)學(xué)分析表明,兩者與擬南芥MAH1/CYP96A15同源性最高。實時熒光定量PCR表明,BnMAH1-1與BnMAH1-2主要在甘藍(lán)型油菜莖、葉、花、及角果中表達(dá),其中在葉片中的表達(dá)量最高,在根系中的表達(dá)量很低,這與角質(zhì)層蠟質(zhì)主要沉積在植株地上部分相一致。BnMAH1-1和BnMAH1-2在無蠟粉材料莖、葉片中幾乎不表達(dá),表明蠟質(zhì)的減少與MAH1的轉(zhuǎn)錄下調(diào)有關(guān)。BnMAH1-1與BnMAH1-2受SA、Me JA、ACC、ABA、Na Cl及干旱脅迫誘導(dǎo)表達(dá),其中BnMAH1-1可能在水分脅迫響應(yīng)中起主要作用。
[Abstract]:In the waxy synthesis pathway of plant, mid-chain alkane hydroxylase- (MAH) catalyzes the hydroxylation of alkanes to form secondary alcohols, which are further oxidized to ketones. In this study, two full-length coding regions of Arabidopsis thaliana P450 dependent enzyme CYP96A15/MAH1 were obtained by means of electronic cloning and RT-PCR techniques. They were named BnMAH1-1 and BnMAH1-2(Gen Bank accession numbers KT795344 and KT79534545, respectively. The length of ORF was 1491 BP, with no intron. The nucleotide and amino acid sequence were 92.4% and 90.9% respectively. The predicted BnMAH1-1 and BnMAH1-2 precursor proteins based on the coding region are polypeptide chains containing 496 amino acid residues. The conserved structure of P450 protein family, P415x R417xK helix E359xx R362C terminal heme binding domain F436xx Gx Rx Cx G445) and the conserved region of oxygen binding band, A / G ~ (309xN) D312x / T / S.NCBI Blast Ns, amino acid sequence multiple weight alignment and phylogenetic analysis showed that, They share the highest homology with Arabidopsis MAH1/CYP96A15. Real-time fluorescence quantitative PCR showed that BnMAH1-1 and BnMAH1-2 were mainly expressed in stems, leaves, flowers, and pods of Brassica napus. The results indicated that the decrease of wax was related to the down-regulation of MAH1 transcription. BnMAH1-1 and BnMAH1-2 were induced by SAMe JACC-ABANa NaCl and drought stress. BnMAH1-1 may play a major role in water stress response.
【作者單位】： 西南大學(xué)農(nóng)學(xué)與生物科技學(xué)院;
【基金】：重慶市基礎(chǔ)與前沿研究計劃項目(cstc2016jcyjA0170) 中央高�；究蒲袠I(yè)務(wù)費專項資金(XDJK2014B037)資助~~
【分類號】：S565.4
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本文編號：1844542