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作物学报 ›› 2025, Vol. 51 ›› Issue (2): 301-311.doi: 10.3724/SP.J.1006.2025.44098

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘薯激酶基因IbHT1的克隆及抗旱性功能鉴定

王语新,陈天羽,翟红,张欢,高少培,何绍贞,赵宁,刘庆昌*   

  1. 农业农村部甘薯生物学与生物技术重点实验室 / 中国农业大学农学院, 北京100193

  • 收稿日期:2024-06-17 修回日期:2024-09-18 接受日期:2024-09-18 出版日期:2025-02-12 网络出版日期:2024-10-11
  • 基金资助:
    本研究由财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-10)资助。

Cloning and characterization of drought tolerance function of kinase gene IbHT1 in sweetpotao

WANG Yu-Xin,CHEN Tian-Yu,ZHAI Hong,ZHANG Huan,GAO Shao-Pei,HE Shao-Zhen,ZHAO Ning,LIU Qing-Chang*   

  1. Key Laboratory of Sweetpotato Biology and Biotechnology, Ministry of Agriculture and Rural Affairs / College of Agronomy & Biotechnology, China Agricultural University, Beijing 100193, China
  • Received:2024-06-17 Revised:2024-09-18 Accepted:2024-09-18 Published:2025-02-12 Published online:2024-10-11
  • Supported by:
    This study was supported by the China Agriculture Research System of MOF and MARA (CARS-10).

摘要:

HT1 (HIGH LEAF TEMPERATURE 1)属于蛋白激酶,在模式植物拟南芥中主要参与气孔运动,但其在甘薯(Ipomoea batatas (L.) Lam.)中的作用未见相关报道。本研究从甘薯品系徐薯55-2中克隆得到IbHT1基因,其CDS全长1140 bp,编码379个氨基酸。IbHT1蛋白具有一个保守的STKc_MAP3K_Like蛋白激酶结构域,预测分子量大小43.07 kD,等电点8.83。其基因组全长2796 bp,含有3个外显子和2个内含子。IbHT1蛋白定位于细胞膜上,其蛋白全长无转录激活活性。IbHT1基因受20% PEG-6000诱导下调表达。过表达IbHT1基因减弱了甘薯植株的抗旱性,而RNA干扰该基因显著增强了甘薯植株的抗旱性。通过酵母筛库筛选到与IbHT1蛋白互作的10个蛋白,由此推测,IbHT1蛋白激酶可能通过与这些蛋白相互作用从而共同参与甘薯抗旱性的调控。

关键词: 甘薯, IbHT1, RNA干扰, 抗旱性, 互作蛋白

Abstract:

HT1 (HIGH LEAF TEMPERATURE 1) is a protein kinase known for its role in regulating stomatal movement in Arabidopsis. However, its function in sweetpotato has not been reported. In this study, the IbHT1 gene was cloned from the sweetpotato line Xushu 55-2. The full-length CDS of IbHT1 is 1140 bp, encoding a 379-amino acid protein that contains a conserved STKc_MAP3K_Like domain, with a predicted molecular weight of 43.07 kD and an isoelectric point (pI) of 8.83. The genomic sequence of IbHT1 spans 2796 bp, comprising 3 exons and 2 introns. Subcellular localization analysis revealed that the IbHT1 protein is localized to the cell membrane, and yeast assays confirmed it lacks transactivation activity. Expression of IbHT1 was down-regulated in response to 20% PEG-6000 treatment. Overexpression of IbHT1 significantly reduced drought tolerance in sweetpotato, while RNA interference (RNAi) of IbHT1 markedly enhanced drought tolerance. Additionally, 10 proteins interacting with IbHT1 were identified through yeast library screening. These findings suggest that IbHT1 may regulate drought tolerance in sweetpotato by interacting with other proteins.

Key words: sweetpotato, IbHT1, RNA interference, drought tolerance, interaction protein

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