水稻OsPIN1d基因克隆与转化及其在水稻根负向光性中的作用
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

浙江省自然科学基金(LY14C130003);


OsPIN1d Gene Clone and Its Function Analysis in Negative Phototropism of Rice Roots
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    生长素极性运输输出载体OsPIN1基因家族可能参与调控水稻根负向光性,其中OsPIN1aOsPIN1b参与水稻根负向光性已得到证实。为了研究OsPIN1d基因与水稻根负向光性形成的关系,依据GenBank数据库中OsPIN1d (Accession number: BR000830) 的核苷酸序列,设计特异性引物,通过RTPCR从水稻根尖的cDNA中扩增出完整的OsPIN1d基因全长。生物信息学分析表明,OsPIN1d 的序列全长为1 497 bp,编码554个氨基酸,GC含量为64.08%;氨基酸序列多重比对及系统发育树构建表明,OsPIN1d与水稻OsPIN1b、玉米OsPIN4以及拟南芥AtPIN2、OsPIN1cOsPIN1aAtPIN1等基因的遗传距离较近。通过构建融合超表达载体pCAMBIA1301OsPIN1d∷GFP,转化并获得其转基因水稻,经RTPCR检测和GUS染色结果显示,外源片段已成功整合到水稻基因组内,并在根部高效表达;受单侧光照射后,转基因水稻种子的根负向光性明显大于野生型,且向光侧OsPIN1dGFP荧光密度明显弱于背光侧。研究表明,OsPIN1d参与了水稻根负向光性的IAA和NAA的运输,从而促进了根负向光性的形成。

    Abstract:

    OsPIN1a and OsPIN1b of auxin efflux carrier OsPIN1 family genes were demonstrated in regulating negative phototropism of rice roots. To study relationship between OsPIN1d and negative phototropism of rice roots, we amplified the complete open reading frame of OsPIN1d through reverse transcriptasepolymerase chain reaction (RTPCR) based on sequence deposited in GenBank (Accession number: BR000830). Bioinformatics analysis showed that OsPIN1d was consisted of 1 497 bp, coding 554 amino acids (AA), and GC content was 64.08%. AA sequence comparison and phylogenic tree indicated that OsPIN1d gene belongs to plant PINs, which is closed to OsPIN1b in rice, OsPIN4 in maize, AtPIN2, OsPIN1c, OsPIN1a and AtPIN1 in Arabidopsis. Fusion expression vector pCAMBIA1301OsPIN1d∷GFP containing OsPIN1d gene was constructed to obtain T2 generation transgenic rice. RTPCR molecular detection and βglucuronidase staining showed that the target construct was integrated into genome of rice, mainly locating in root. Irradiated with unilateral light, the negative phototropic curvatures of transgenic rice seed roots were higher than those of wild type. Moreover, OsPIN1dGFP in shaded side was significantly higher than that in irradiated side in elongation zone of transgenic rice root tip. It was indicated that OsPIN1d is involved in transportation of IAA and NAA, thus promoting formation of negative phototropism in rice roots.

    参考文献
    相似文献
    引证文献
引用本文

梁国斌,胥华伟,方 群,等.水稻OsPIN1d基因克隆与转化及其在水稻根负向光性中的作用[J].西北植物学报,2017,37(10):1896-1903

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2017-12-01
  • 出版日期:

微信公众号二维码

手机版网站二维码