中国农业科学 ›› 2008, Vol. 41 ›› Issue (3): 636-642 .doi: 10.3864/j.issn.0578-1752.2008.03.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦细胞分裂素氧化/脱氢酶基因(TaCKX5)的克隆及其染色体定位

张 磊,张宝石,周荣华,孔秀英,高丽峰,贾继增   

  1. 沈阳农业大学农学院
  • 收稿日期:2007-01-05 修回日期:1900-01-01 出版日期:2008-03-10 发布日期:2008-03-10
  • 通讯作者: 贾继增

Isolation and chromosomal localization of cytokinin oxidase/dehydrogenase gene (TaCKX5) in wheat

  

  1. 沈阳农业大学农学院
  • Received:2007-01-05 Revised:1900-01-01 Online:2008-03-10 Published:2008-03-10

摘要: 【目的】克隆小麦的TaCKX基因并对其序列进行生物信息学分析,明确TaCKX基因在小麦基因组中的分布情况,以期为今后深入研究小麦CKX基因以及利用该基因进行小麦重要农艺性状的遗传改良奠定基础。【方法】采用同源克隆结合BAC文库筛选的方法克隆基因,通过小麦缺体-四体对其进行染色体定位。【结果】从普通小麦中国春中分离得到TaCKX5基因的gDNA和cDNA序列。分析表明,TaCKX5基因的开放阅读框为1 596 bp,编码531个氨基酸,含有CKX基因家族典型的功能位点FAD-binding domain,预测属于分泌蛋白,并具有糖基化位点。使用中国春缺体-四体将TaCKX5定位于小麦第三同源群。系统发育分析表明,CKX基因在植物中较为保守,禾谷类作物中直向同源的CKX基因很可能具有相似的特性与功能。【结论】分离获得了普通小麦TaCKX5基因全长,TaCKX5分布于小麦染色体3A、3B、3D上,与水稻OsCKX5基因直向同源。

关键词: 小麦, 细胞分裂素氧化/脱氢酶, 基因克隆, 染色体定位

Abstract: 【Objective】Genetic manipulation of CKX activity holds the promise of improving agricultural traits in crops.【Method】and【Result】In this study, a strategy of homologous cloning combined with BAC library screening was used to isolate TaCKX5 gene from wheat. Sequence analysis of TaCKX5 revealed that TaCKX5 contains a FAD-binding domain which is well conserved in the N-terminal halves of all known CKX sequences. Using TargetP program, TaCKX5 was predicted to be a secretion pathway protein, and one N-glycosylation site (NFTE) was also found with NetNGly program, implied that TaCKX5 protein may be modified posttranslationally by glycosylation. Using nulli-tetrasomic stocks, TaCKX5 was located on wheat homoeologous group 3 chromosomes.【Conclusion】TaCKX5 was very closely related to its orthologous CKX gene (OsCKX5) in rice, so they might have similar characters and functions.

Key words: Wheat, Cytokinin Oxidase/dehydrogenase, Cloning, Chromosomal Localization.