中间锦鸡儿CiPUB22基因克隆与功能分析
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

内蒙古自治区科技创新引导项目(KCBJ2018012)


Cloning and Functional Characterization of CiPUB22 from Caragana intermedia
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:

    中间锦鸡儿(Caragana intermedia)耐旱、耐寒、耐盐碱,是西北干旱地区的重要固沙灌木,筛选其优良抗逆境基因,可以作为林草基因工程的基因源。该研究在中间锦鸡儿干旱胁迫转录组文库中找到1条CiPUB22 (plant Ubox 22)基因的cDNA全长序列,CiPUB22基因包括1 260 bp开放阅读框,编码419个氨基酸。实时荧光定量PCR结果表明,在脱水、盐和ABA处理1 h后CiPUB22基因表达量上升并达到最高水平,分别为对照表达量的12倍、35倍和7倍,干旱处理后12 d达到最高值,为对照的2.5倍,表明CiPUB22的转录水平受非生物胁迫诱导。构建CiPUB22基因的过表达载体并转化野生型拟南芥,对转基因纯合体株系抗逆性分析发现,在150 mmol/L NaCl、1 μmol/L ABA和400 mmol/L甘露醇处理下,过表达株系的萌发率均低于野生型,说明过表达CiPUB22基因降低了拟南芥在种子萌发过程中对盐和渗透胁迫的耐受性。

    Abstract:

    Caragana intermedia is an important sandfixing shrub in northwest arid area with drought, cold and salinealkali resistance. Excellent stress resistant candidate genes screened from it could provide good gene resources for genetic engineering of forest and forage species. In this study, the full length cDNA sequence of CiPUB22 (plant Ubox 22) had been mined from the transcriptome library constructed in our laboratory using drought stressed seedlings of C. intermedia. The CiPUB22 gene contained 1 260 bp open reading frame (ORF) which encoding 419 amino acids. Quantitative real time PCR analysis showed that the expression levels of CiPUB22 were significantly increased and reached the highest level, which was 12, 35 and 7 times of the control at 1 h after dehydration, NaCl and ABA treatments, respectively. After drought treatment, the relative expression of CiPUB22 was significantly increased and reached the highest level at 12 d, which was 2.5 times more than that of the control. The results indicated that the transcription of CiPUB22 was induced by abiotic stress. Furthermore, expression vector pCanGCiPUB22 was constructed and transformed into the wildtype Arabidopsis. Homozygous transgenic Arabidopsis overexpressing CiPUB22 were generated for further analysis. Seed germination assay showed that CiPUB22 transgenic lines germinated slower than that of wildtype on medium containing 150 mmol/L NaCl, 400 mmol/L mannitol or 1 μmol/L ABA. These results indicated that CiPUB22 decreased tolerance of the transgenic Arabidopsis to salt and osmotic treatments during seed germination.

    参考文献
    相似文献
    引证文献
引用本文

岳文冉,于秀敏,岳俊燕,等.中间锦鸡儿CiPUB22基因克隆与功能分析[J].西北植物学报,2018,38(11):1961-1967

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: 2018-12-20
  • 出版日期:

微信公众号二维码

手机版网站二维码