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园艺学报 ›› 2020, Vol. 47 ›› Issue (8): 1521-1529.doi: 10.16420/j.issn.0513-353x.2019-0935

• 研究论文 • 上一篇    下一篇

枸杞Lb_PPR1的克隆及其在不同育性品种中的表达分析

管翠萍,杨亚珺,石 晶*   

  1. 宁夏大学生命科学学院,西部特色生物资源保护与利用教育部重点实验室,银川 750021
  • 出版日期:2020-08-25 发布日期:2020-08-25
  • 基金资助:
    宁夏自然科学基金项目(2019AAC03014)

Cloning and Expression Analysis of Lb_PPR1 Gene in Different Lycium barbarum Fertility Varieties

GUAN Cuiping,YANG Yajun,and SHI Jing*   

  1. College of Life Science,Ningxia University;Western Biological Resources Protection and Utilization Lab of National Education Ministry,Yinchuan 750021,China
  • Online:2020-08-25 Published:2020-08-25

摘要: 三角状五肽重复(Pentatricopeptide repeats,PPR)蛋白定位于多种细胞器中,参与细胞核和细胞器中特异单链RNA的转录后修饰和编辑,在植物生长发育的多个阶段均发挥着重要的作用。分别从枸杞(Lycium barbarum)雄性可育系‘宁杞1号’和不育系‘宁杞5号’中克隆了Lb_PPR1基因,并对其编码蛋白质进行理化性质、亚细胞定位、保守结构域、蛋白结构以及系统进化等方面进行预测分析。结果显示,‘宁杞1号’和‘宁杞5号’中Lb_PPR1基因的开放阅读框均为1 977 bp,编码658个氨基酸,但其中存在20个碱基和14个氨基酸的差异。Lb_PPR1蛋白包含14个串联重复的PPR保守基序,属于PLS家族,该蛋白定位在细胞膜和细胞质。二级、三级结构预测显示该蛋白以α螺旋为主。同源进化分析得出Lb_PPR1蛋白与同属茄科的辣椒和烟草PPR蛋白亲缘关系最近。利用实时荧光定量PCR检测Lb_PPR1在枸杞不同组织器官及不同发育阶段花药中的表达特性。结果显示,Lb_PPR1在枸杞不同组织中均有表达,但在可育系花蕾中表达量最高,不育系‘宁杞5号’各组织中表达量均显著低于可育系‘宁杞1号’。以上结果表明,Lb_PPR1基因可能与枸杞花药发育有关。

关键词: 枸杞, Lb_PPR1, 生物信息, 表达模式

Abstract: The pentatricopeptide repeats(PPR)proteins involve in post-transcriptional modifications and editing of specific single-stranded RNAs in the nucleus and organelles via their localization in various organelles,and they play important roles in series events of plant growth and development. In this study,Lb_PPR1 gene was cloned from the fertile line‘Ningqi 1’and the male-sterile line‘Ningqi 5’. Bioinformatics analysis was used to predict the physico-chemical properties,subcellular localization,conserved domain,protein structure and phylogenetic evolution of Lb_PPR1 encoded protein. The results indicated that Lb_PPR1 gene contained a 1 977 bp open reading frame encoding 658 amino acids in both ‘Ningqi 1’and ‘Ningqi 5’,but there were 20 bp and 14 aa differences between the two cultivars. Lb_PPR1 protein was predicted to have 14 tandem repeats,belonged to PLS family,and its subcellular localization was in cell membrane and cytoplasm. Secondary and tertiary structure predictions showed that Lb_PPR1 protein was dominated by α-helix. Phylogenetic analysis showed that Lb_PPR1 had the closest relationship with PPR in Capsicum annuum and Nicotiana attenuate. Quantitative real-time PCR(qRT-PCR)was performed to determine the expression pattern of Lb_PPR1 gene in different organs and different developmental stages of anther. The results showed that Lb_PPR1 was expressed in different tissues of Lycium barbarum,but highest in the fertile line’s flower buds,its expression in the male-sterile line‘Ningqi 5’were all significantly lower than the fertile line‘Ningqi 1’. The above results indicate that Lb_PPR1 gene maybe related to the development of flower organs in Lycium barbarum.

Key words: Lycium barbarum, Lb_PPR1, bioinformatics analysis, gene expression pattern

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