本文選題：芍藥 + PlGAox��； 參考：《植物生理學(xué)報(bào)》2017年04期

【摘要】：以芍藥‘大富貴’芽為試材,采用RT-PCR結(jié)合c DNA末端快速擴(kuò)增(RACE)技術(shù),克隆得到GA20ox基因的c DNA全長,命名為Pl GA20ox(Gen Bank登錄號(hào)為KU886552)。該基因全長1 351 bp,開放閱讀框1 146 bp,共編碼381個(gè)氨基酸,含有高度保守的20G-Fe(I I)-Oxy蛋白結(jié)構(gòu)域、Fe2+結(jié)合位點(diǎn)(His-247、Asp-249和His-303)和2-酮戊二酸結(jié)合位點(diǎn)(Arg-313和Ser-315)。Pl GA20ox蛋白分子量為43 209.1 Da,為穩(wěn)定蛋白,無跨膜結(jié)構(gòu)域,無信號(hào)肽,屬于C19-GAoxs。氨基酸序列同源性及系統(tǒng)進(jìn)化樹分析表明:Pl GA20ox與牡丹Ps GA20ox同源性高達(dá)96%,親緣關(guān)系最近。采用農(nóng)桿菌介導(dǎo)法將Pl GA20ox基因于本生煙葉片中瞬時(shí)表達(dá),觀察顯示:Pl GA20ox蛋白定位于細(xì)胞質(zhì)中。該基因在芍藥各器官中均有表達(dá),其中,在芽中表達(dá)最高,其次花瓣,在根、萼片、葉片和莖中表達(dá)微弱。實(shí)時(shí)熒光定量PCR和高效液相色譜(HPLC)結(jié)果顯示:在低溫解除芍藥芽內(nèi)休眠進(jìn)程中,Pl GA20ox表達(dá)水平呈先上升后總體下降趨勢,且與內(nèi)源GA3含量呈顯著正相關(guān)(r=0.901*);外施赤霉素會(huì)明顯增加內(nèi)源GA3含量,但同時(shí)抑制Pl GA20ox的表達(dá),說明Pl GA20ox調(diào)控GA3的合成,同時(shí)受植物體內(nèi)活性GA3含量的負(fù)反饋調(diào)節(jié)。
[Abstract]:Using Paeonia lactiflora 'Dafugui' bud as test material, the full length of c DNA of GA20ox gene was cloned by RT-PCR and rapid amplification of c DNA terminal. The full length of c DNA was named Pl GA20ox(Gen Bank accession number KU886552. The total length of the gene is 1 351 BP, open reading frame 1 146 BP, encoding 381 amino acids, containing highly conserved 20G-Fe(I I)-Oxy domain His-247 Asp-249 and His-303), 2-ketoglutaric acid binding site Arg-313 and Ser-315).Pl GA20ox protein with molecular weight of 43 209.1 Daa, which is a stable protein. No transmembrane domain, no signal peptide, belonging to C19-GAoxs. The analysis of amino acid sequence homology and phylogenetic tree showed that the homology of the amino acid sequence and the peony Ps GA20ox was 96%, and the phylogenetic relationship was the most recent. Using Agrobacterium tumefaciens mediated expression of Pl GA20ox gene in tobacco leaves, it was observed that the protein of Pl GA20ox was located in the cytoplasm. The gene was expressed in all organs of Paeonia lactiflora, the highest in bud, the second in petals, and weak in roots, sepals, leaves and stems. The results of real-time fluorescence quantitative PCR and high performance liquid chromatography (HPLC) showed that the expression of Pl GA20ox increased first and then decreased during the process of releasing dormancy of Paeonia lactiflora at low temperature. Gibberellin significantly increased the content of endogenous GA3, but inhibited the expression of Pl GA20ox at the same time, indicating that Pl GA20ox regulated the synthesis of GA3 and was negatively regulated by the content of active GA3 in plant.
【作者單位】： 山東農(nóng)業(yè)大學(xué)林學(xué)院;
【基金】：山東省自然科學(xué)基金(ZR2014CM028)~~
【分類號(hào)】：Q943.2;S682.12

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本文編號(hào)：1914938