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热带作物学报 ›› 2020, Vol. 41 ›› Issue (5): 978-984.doi: 10.3969/j.issn.1000-2561.2020.05.018

• 生物技术与组织培养 • 上一篇    下一篇

辣椒胞质雄性不育系CaNAD9线粒体基因的克隆及表达分析

杨婷玉,邵贵芳,张水,王姣,张靖柔,赵凯,邓明华()   

  1. 云南农业大学园林园艺学院,云南昆明 650201
  • 收稿日期:2019-08-01 修回日期:2019-09-02 出版日期:2020-05-25 发布日期:2020-06-15
  • 通讯作者: 邓明华 E-mail:dengminghua2013@163.com
  • 作者简介:杨婷玉(1994—),女,硕士研究生,研究方向:蔬菜遗传育种。
  • 基金资助:
    国家自然科学基金(31760575);云南省科技计划重点项目(2018BB020)

Cloning and Expression Analysis of Mitochondrial Gene of Cytoplasmic Male Sterile Line CaNAD9 in Pepper (Capsicum annuum L.)

YANG Tingyu,SHAO Guifang,ZHANG Shui,WANG Jiao,ZHANG Jingrou,ZHAO Kai,DENG Minghua()   

  1. College of Horticulture and Landscape, Yunnan Agricultural University, Kunming, Yunnan 650201, China
  • Received:2019-08-01 Revised:2019-09-02 Online:2020-05-25 Published:2020-06-15
  • Contact: DENG Minghua E-mail:dengminghua2013@163.com

摘要:

为了深入研究辣椒胞质雄性不育与能量代谢之间的关系,本研究以辣椒胞质雄性不育系9704A和同型保持系9704B为实验材料,克隆了线粒体基因组CaNAD9基因,比较了其一级结构在2种材料间的差异,利用实时荧光定量PCR技术分析该基因在保持系9704B不同组织中的表达模式,以及胞质雄性不育系9704A与同型保持系9704B花蕾不同发育时期的表达差异。结果表明:通过对双向测序进行分析,辣椒CaNAD9基因CDS长度为573 bp;生物信息学分析表明CaNAD9基因编码190个氨基酸残基;CaNAD9基因转录本在不育系和保持系中均未发生RNA编辑;辣椒CaNAD9基因在不同组织中的表达量存在差异,且差异较为明显,其中在种子中的表达量最高,在胎座中的表达量最低;辣椒CaNAD9基因的表达量在胞质雄性不育系与保持系花蕾不同发育阶段中存在一定差异,在花粉母细胞减数分裂期辣椒胞质雄性不育系与同型保持系中的表达差异最为显著。这种差异可能会使雄性不育系的能量代谢出现异常,从而导致辣椒雄性不育。

关键词: 辣椒, CaNAD9, 胞质雄性不育, 能量代谢, 基因表达

Abstract:

In order to further study the relationship between the cytoplasmic male sterility and energy metabolism in pepper, the cytoplasmic male sterile line 9704A and homologous maintainer line 9704B were used as the experimental materials to clone the mitochondrial genome CaNAD9 gene and compare its primary structure. The difference between the two materials was analyzed by real-time quantitative PCR to analyze the expression pattern of the gene in different tissues of the maintainer line 9704B and the expression differences of the buds of the cytoplasmic male sterile line 9704A and the homologous maintainer line 9704B. The CDS length of the CaNAD9 consisted of 573 bp, which encoding 190 amino acids. RNA editing of the CaNAD9 transcript did not happened in both 9704A and 9704B. The expression of CaNAD9 in pepper was different in different tissues with the highest expression in seeds and the lowest in fetuses. There were some differences in different developmental stages of the flower buds, and the difference in the expression of the cytoplasmic male sterile lines and the homologous maintainers in the meiotic stage of pollen mother cells was the most significant. This difference may cause abnormalities in the energy metabolism of the male sterile line, leading to male sterility in the pepper.

Key words: pepper (Capsicum annuum L.), CaNAD9, cytoplasmic male sterility, energy metabolism, gene expression

中图分类号: 

  • S188