海岛棉WUSCHEL基因(GbWUS)的克隆与表达模式分析

Coning and expression pattern analysis of WUSCHEL gene from Gossypium barbadense( GbWUS)

[目的]分离海岛棉的WUSCHEL基因(Gb WUS),检测Gb WUS基因在海岛棉体细胞胚发生过程中的表达模式。[方法]利用同源序列法结合RACE技术的策略克隆Gb WUS基因,通过半定量RT-PCR分析Gb WUS基因的表达模式。[结果]获得了全长1 011 bp的Gb WUS基因c DNA序列,包括870bp的开放阅读框(ORF),编码289个氨基酸。与来源于其他物种WUS同源基因的进化树分析表明,Gb WUS与雷蒙德氏棉WUS基因具有98%的同源性,而与其他物种来源的WUS同源基因有明显的种属特异性。Gb WUS基因在海岛棉的胚性愈伤组织中表达,而在非胚性愈伤、球型胚、子叶胚阶段的细胞中不表达。[结论]Gb WUS基因属于WUSCHEL/WOX家族,其mRNA在胚性愈伤组织中特异表达。

[ Objective ] Isolating WUS gene from G. barbadense, and checking the expression pattern of GbWUS in the G. bar- badense somatic embryogenesis process. [ Methods] GbWUS gene was cloned with homology sequence cloning and RACE tech- nology. And expression pattern of GbWUS was checked through semi -quantitative RT -PCR. [ Results ] The 1 011 bp GbWUS cDNA was got including a 870 bp open reading frame （ ORF）, which encoded 289 amino acid. Phylogenetic tree analysis showed GbWUS have 98% homology with WUS gene in Gossypium raimondii but a distinctive difference with WUS homologs from the other origins. And GbWUS was detected to express in embryonic callus, but not in the non -embryonic callus, globular embryo or leaf- embryo. [ Conclusion] GbWUS belong to a member of WUSCHEL/WOX gene family, and expressed in the embryonic callus.