生物技术通报 ›› 2020, Vol. 36 ›› Issue (11): 21-29.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0510

• 研究报告 • 上一篇    下一篇

水稻籽粒低镉蛋白LCD互作蛋白的筛选与鉴定

郝小花1,2(), 戴佳利2, 暨文劲2, 黄丹2, 李东屏2, 田连福2()   

  1. 1.湖南文理学院生命与环境科学学院,常德 415000
    2.湖南师范大学作物生命科学学院 作物不育资源创新与利用湖南省重点实验室,长沙 410081
  • 收稿日期:2020-04-30 出版日期:2020-11-26 发布日期:2020-11-20
  • 作者简介:郝小花,女,博士,研究方向:植物生态学;E-mail: 14771417@qq.com
  • 基金资助:
    国家自然科学基金项目(31570316);湖南省教育厅项目(15B160);湖南省教育厅项目(17C0974);湖南省自然科学基金项目(13JJ3116);湖南省研究生创新性项目(CX2018B300);湖南文理学院应用特色学科-生物学,湖南省生物发育工程及新产品研发协同创新中心(20134486)

Screening and Identification of LCD-interacting Proteins in Rice

HAO Xiao-hua1,2(), DAI Jia-li2, JI Wen-jin2, HUANG Dan2, LI Dong-ping2, TIAN Lian-fu2()   

  1. 1. College of Life and Environmental Sciences,Hunan University of Arts and science,Changde 415000
    2. College of Life Science,Hunan Normal University,Hunan Province Key Laboratory of Crop Sterile Germplasm Resource Innovation and Application,Changsha 410081
  • Received:2020-04-30 Published:2020-11-26 Online:2020-11-20

摘要:

水稻低镉基因(Low cadmium,LCD)参与水稻籽粒镉(Cd)的积累,筛选与LCD互作的蛋白有利于揭示LCD介导的籽粒Cd积累调控的分子机制。以粳稻日本晴为试验材料构建酵母双杂交膜系统和核系统cDNA文库,以LCD为诱饵蛋白,分别进行膜系统和核系统筛选,获得29个与LCD互作的候选蛋白,并对这些基因进行gene ontology(GO)注释分析。进一步通过一对一互作验证,获得21个互作蛋白,包括膜系统蛋白16个,核系统蛋白5个,其中,6个蛋白与物质转运相关,5个蛋白与植物耐逆相关,4个蛋白与植物发育响应相关,6个蛋白与细胞生理代谢相关。

关键词: 水稻, LCD, 低镉, 酵母双杂交, 互作蛋白

Abstract:

Low cadmium(LCD)gene is involved in grain Cd accumulation of rice. To screen LCD-interacting proteins is beneficial for revealing the molecular mechanism of LCD-mediated grain Cd accumulation and regulation. Using Oryza sativa ssp. japonica cv. Nipponbare as experimental material,the cDNA libraries of yeast two hybrid membrane system and nuclear system were constructed,and LCD was used as bait protein to screen the membrane system and nuclear system respectively. A total of 29 genes were obtained,and the gene ontology(GO)analyses were conducted to annotate their molecular function. Further,these genes were cloned and one-to-one interaction verification with LCD was performed. Finally,a total of 21 LCD-interacting proteins including 16 from membrane system and 5 from nuclear system were identified. Among them,6 proteins were related to the tolerance and transport of metal ions,5 were related to the stress responses,4 were related to plant development responses,and the rest 6 were related to cellular and physiological metabolisms.

Key words: rice, LCD, low cadmium, yeast two-hybrid(Y2H), interacting protein