FaesAP2B基因在甜荞长雌蕊长雄蕊突变体lpls的表达分析

Expression Analysis of the FaesAP2B in Mutant Buckwheat with Long Pistil and Long Stamen

为弄清甜荞(Fagopyrum esculentum Moench.)长雌蕊长雄蕊突变体lpls花和籽粒发育调控的分子机制,从甜荞中克隆出1个长1788 bp的AP2同源基因的cDNA序列,命名为FaesAP2B(GenBank登录号为MK290847.1)。序列结构分析表明:FaesAP2B基因包含1个长1380 bp的完整开放阅读框(Open Reading Frame,ORF),编码1个由459个氨基酸残基组成的AP2/ERF家族转录因子,该转录因子含有2个高度保守的AP2结构域,第1个AP2结构域前还存在1个由10个氨基酸残基组成的核定位信号区。用qPCR检测FaesAP2B基因在甜荞lpls突变体根、茎、幼叶、花被片、雄蕊、雌蕊以及发育4 d的果实共7种器官中表达的组织特异性显示:FaesAP2B在甜荞突变体lpls营养组织和生殖结构中均有表达,但其在花器官和果实等生殖结构中的表达量明显高于营养组织,且在雄蕊中的表达量最高,极显著高于其在其他6种组织中的表达量(LSD,P<0.01),同时,FaesAP2B在花被片、雌蕊和发育4 d的果实中的表达量均极显著高于其在根、茎和叶等营养器官中的表达量(LSD,P<0.01),但该基因在其根、茎、叶间的表达量无显著性差异。推测该基因可能主要参与调控甜荞lpls突变体花和果实的发育。

In order to uncover the molecular mechanism of flower and grain development of buckwheat(Fagopyrum esculentum Moench.)with long pistil and long stamen mutant‘lpls’of a 1788 bp long AP2 homologous cDNA sequence was cloned from F.esculentum Moench.and designated as FaesAP2B(GenBank accession number is MK290847.1).By sequence analysis,FaesAP2B gene contains a full Open Reading Frame with 1380 bp long,and encodes an AP2/ERF family transcription factor consisting of 459 amino acid residues.The FaesAP2B transcription factor contains two highly conserved AP2 domains,with a nuclear localization signal region consisting of ten amino acid residues lying before the first AP2 domain.qPCR was performed to detect FaesAP2B gene expression in root,stem,leaf,perianth,stamen,pistil and juvenile fruit in four-day buckwheat mutant lpls,respectively.The FaesAP2B is expressed in both vegetative tissues and reproductive structures.However,the expression level of FaesAP2B in reproductive organs such as floral organs and fruits is significantly higher than that in vegetative tissues.In addition,the expression level of FaesAP2B in stamens is the highest among seven different organs(LSD,P<0.01).And the expression level of FaesAP2B in the perianth,pistil and four-day fruit was significantly higher than that in the vegetative organs such as roots,stems and leaves(LSD,P<0.01),but there was no significant difference in the expression of the gene between roots,stems and leaves.Our data suggested that FaesAP2B may be mainly involved in regulating flower and fruit development in the buckwheat mutant‘lpls’.