甘蓝BoQRT3基因的克隆和表达分析

Cloning and Expression Analysis of BoQRT3 Gene in Brassica oleracea

QRT3(QUARTET3)基因在植物花粉正常发育过程中发挥着重要作用。为了探究甘蓝QRT3的功能,利用同源克隆的方法得到了BoQRT3基因的DNA和编码序列的全长,并对其进化和表达模式进行了研究。序列分析表明,BoQRT3共有2123个核苷酸,包括3个外显子和2个内含子;编码序列为1428 bp,编码475个氨基酸;蛋白质相对分子量和等电点分别为51.02 kD和5.77,包含一个长度为27个氨基酸残基的信号肽,定位在膜外,属于一种分泌蛋白。实时荧光定量PCR分析表明,BoQRT3在花序中特异高表达,在根中也有相对较高的表达,但在其他组织、器官中几乎不表达。原位杂交结果表明,BoQRT3只在甘蓝花粉发育的四分体时期的绒毡层细胞中特异性表达。试验结果为深入研究BoQRT3在甘蓝花粉发育中的生物学功能及调控网络提供了理论依据。

QRT3(QUARTET 3)gene plays an important role in the development of pollen.In order to explore the function of BoQRT3 in Brassica oleracea,the full length of DNA and coding sequence of BoQRT3 was obtained by homologous cloning,and its evolution and expression patterns were also studied.Sequence analysis showed that BoQRT3 had a total of 2123 nucleotides,including 3 exons and 2 introns;the coding sequence was 1428 bp,encoding 475 amino acids;the relative molecular weight and isoelectric point of the protein were 51.02 kD and 5.77,respectively.BoQRT3 contained a signal peptide with a length of 27 amino acid residues,located outside the membrane,and was a secreted protein.RT-PCR analysis showed that BoQRT3 was specifically highly expressed in the inflorescences and also had relatively high expression in roots,but it was almost not expressed in other tissues and organs of B.oleracea.In situ hybridization analysis further showed that BoQRT3 was specifically expressed in the tapetum cells during the tetrad stage of pollen development in B.oleracea.Our results provided a theoretical basis for in-depth study of the biological function and regulatory network of BoQRT3 in the pollen development of B.oleracea.