菊花CmPIN1同源基因的克隆與表達(dá)分析
發(fā)布時(shí)間:2018-06-24 04:52
本文選題:菊花 + CmPIN ; 參考:《中國農(nóng)業(yè)大學(xué)學(xué)報(bào)》2016年03期
【摘要】:以切花菊‘神馬’(Chrysanthemum morifolium‘Jinba’)為材料,利用RACE技術(shù)與同源克隆方法獲得菊花CmPIN1基因全長,通過qRT-PCR技術(shù)比較CmPIN1在不同組織器官的表達(dá),同時(shí)對(duì)CmPIN1響應(yīng)外施NAA和去頂進(jìn)行研究。結(jié)果表明:1)CmPIN1基因ORF全長1 761bp,編碼586個(gè)氨基酸,跨膜域位于蛋白N端與C端;通過蛋白序列比對(duì)分析,菊花CmPIN1蛋白與百日草ZvPIN1蛋白相似性最高;亞細(xì)胞定位結(jié)果顯示CmPIN1位于細(xì)胞膜;2)對(duì)CmPIN1表達(dá)分析表明,該基因在菊花莖部與芽部具有相對(duì)較高的表達(dá)量,同時(shí),在離體莖段中的CmPIN1受到生長素誘導(dǎo);去頂后,CmPIN1表達(dá)量降低,而施加5μmol/L的NAA 6h后CmPIN1表達(dá)恢復(fù);3)對(duì)菊花植株進(jìn)行去頂試驗(yàn)表明,去頂后,近頂端第一個(gè)側(cè)芽內(nèi)CmPIN1基因表達(dá)量優(yōu)先恢復(fù),趨向于代替頂芽成為新的生長素源,以維持主莖中生長素極性運(yùn)輸;當(dāng)主莖中的生長素運(yùn)輸通道再次打開,莖節(jié)中CmPIN1的表達(dá)量均逐漸恢復(fù),其中,在近頂端第一個(gè)節(jié)間中CmPIN1的表達(dá)量恢復(fù)較緩慢?梢奀mPIN1參與菊花主莖中生長素的極性運(yùn)輸,以及頂端優(yōu)勢的維持,間接抑制側(cè)芽伸長。
[Abstract]:The full-length CmPIN1 gene of Chrysanthemum morifolium Jinba' was obtained by race and homologous cloning. The expression of CmPIN1 in different tissues and organs was compared by qRT-PCR, and the response of CmPIN1 to NAA and topping was studied. The results showed that the ORF of the CmPIN1 gene was 1761 BP, encoding 586 amino acids, and the transmembrane domain was located at the N-terminal and C-terminal of the protein, and the similarity between CmPIN1 protein and ZvPIN1 protein of Chrysanthemum chrysanthemum was the highest. The results of subcellular localization showed that CmPIN1 was located on cell membrane 2) and the expression of CmPIN1 was relatively high in stems and buds of chrysanthemum, meanwhile, CmPIN1 was induced by auxin in stem segments in vitro, and the expression of CmPIN1 was decreased after topping. CmPIN1 gene expression in the first lateral bud of the near apical bud recovered preferentially and tended to replace the apical bud as a new auxin source after the ceilings were removed by 5 渭 mol / L NAA for 6 h after the restoration of CmPIN1 expression in the chrysanthemum plants, and the results showed that the expression of CmPIN1 gene in the first lateral bud of the near apical bud recovered preferentially. In order to maintain the polar transport of auxin in the main stem, when the auxin transport channel in the main stem was opened again, the expression of CmPIN1 was gradually restored in the stem nodes, and the expression of CmPIN1 was slowly restored in the first internode near the top of the stem. CmPIN1 was involved in the polar transport of auxin in the main stem of chrysanthemum and the maintenance of apical dominance, which indirectly inhibited the elongation of lateral buds.
【作者單位】: 中國農(nóng)業(yè)大學(xué)農(nóng)學(xué)與生物技術(shù)學(xué)院;
【基金】:國家‘863’計(jì)劃項(xiàng)目(2011AA100208) 農(nóng)業(yè)部“引進(jìn)國際先進(jìn)農(nóng)業(yè)科學(xué)技術(shù)”重點(diǎn)項(xiàng)目(2008-G3)
【分類號(hào)】:S682.11
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本文編號(hào):2060101
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