生物学杂志

生物学杂志

• 研究报告 •    下一篇

毛果杨转录因子PtrWRKY25的克隆及功能验证

  

  1. 1. 中国科学院高原生物适应与进化重点实验室，中国科学院西北高原生物研究所，西宁 810001； 2.中国科学院大学，北京 100039）
  • 出版日期:2015-12-18 发布日期:2015-12-18
  • 通讯作者: 周党卫,博士,副研究员,主要:事高山植物适应于进化研究,E-mail：dangweizhou@sina.com。
  • 作者简介:尹佳,汉族,硕士研究生,主要从事植物遗传多样性研究
  • 基金资助:
    国家自然基金, 31070208

Molecular Cloning and Characterization of Transcription Factor PtrWRKY25 from Populus trichocarpa

  1. （1. Key laboratory of Adaption and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001; 2. University of Chinese Academy of Sciences, Beijing 100039,China）
  • Online:2015-12-18 Published:2015-12-18

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摘要： 通过RTPCR的方法获得毛果杨（Populus trichocarpa）PtrWRKY25（Potri.007G078200.1）。经氨基酸比对分析，毛果杨PtrWRKY25蛋白与拟南芥（Arabidopsis thaliana）AtWRKY12、葡萄（Vitis vinifera）VvWRKY2以及苜蓿（Medicago truncatula)MtSTP 的WRKY结构域有较高同源性。利用荧光定量及半定量检测PtrWRKY25组织表达量发现该基因在茎中表达量最高，其次为韧皮部和木质部。植物组织切片间苯三酚染色发现PtrWRKY25转基因株系次生壁较野生型也有减少的趋势。检测农杆菌介导在毛白杨中超量表达的PtrWRKY25转基因株系木质素含量发现，相较于与野生型，转基因植株中木质素含量减少约1倍，表明PtrWRKY25可能主要在木质素合成途径中发挥作用。

Abstract: A complete fragment representing the PtrWRKY25 (Potri.007G078200.1), was isolated from Populus trichocarpa. by reverse transcription PCR(RT-PCR). Displaying significant similarity with other WRKY factors in Arabidopsis’s AtWRKY12, Vitis vinifera’s VvWRKY2 and Medicago truncatula’s MtSTP gene. The expression profiles of PtrWRKY25 showed the highest transcript level in stem and relatively low level in phloem and xylem. Plant tissue sections between phloroglucinol staining found PtrWRKY25 transgenic lines compared with wild-type secondary wall is also a decreasing trend. To investigate the in vivo function, PtrWRKY25 was expressed under control of the CaMV35S promoter in Populus trichocarpa. by Agrobacterium-mediated transformation. About 1 time lower accumulation of lignin was detedted in transgenic plants harboring PtrWRKY25, coincide with the toluidine blue staining of stem. Our findings suggested that PtrWRKY25 might play more important roles in biosynthesis of lignin.