华北农学报 ›› 2024, Vol. 39 ›› Issue (3): 195-207. doi: 10.7668/hbnxb.20194623

所属专题: 油料作物 植物保护 生物技术 热点论文

• 资源环境·植物保护 • 上一篇    下一篇

大豆抗胞囊线虫锌指蛋白基因GmC2H2-2like克隆与初步功能分析

曲硕, 刘芳, 宋庚晨, 胡世豪, 房瑶瑶, 赵雪, 韩英鹏   

  1. 东北农业大学 农学院,农业农村部东北大豆生物学与遗传育种重点实验室,黑龙江 哈尔滨 150030
  • 收稿日期:2024-01-16 出版日期:2024-06-28
  • 通讯作者:
    韩英鹏(1978—),男,黑龙江哈尔滨人,教授,博士,博士生导师,主要从事大豆基因组学遗传育种研究。
  • 作者简介:

    曲 硕(1994—),男,吉林长春人,在读博士,主要从事大豆作物遗传育种研究。

  • 基金资助:
    黑龙江省重点基金项目(ZD2022C002); 黑龙江省重点研发项目(JD22A015); 国家自然科学基金项目(U22A20473); 国家现代农业岗位体系项目(CARS-04-PS07); 东北农业大学科研项目(NEAU2023QNLJ-003)

Soybean Cyst Nematode Resistant Zinc Finger Protein Gene GmC2H2-2like Cloning and Preliminary Functional Analysis

QU Shuo, LIU Fang, SONG Gengchen, HU Shihao, FANG Yaoyao, ZHAO Xue, HAN Yingpeng   

  1. College of Agriculture,Northeast Agricultural University,Northeast Key Laboratory of Soybean Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs,Harbin 150030,China
  • Received:2024-01-16 Published:2024-06-28

摘要:

为了研究锌指蛋白C2H2在大豆胞囊线虫病(SCN)胁迫应答中起到的重要作用,为进一步研究抗大豆胞囊线虫奠定重要基础。利用东农L-10(抗SCN)、黑农37(感SCN)接种SCN3根系转录组数据,筛选出差异表达基因GmC2H2-2like。倒置荧光显微镜下观察该GmC2H2-2like定位情况,对该基因进行生物信息学分析,克隆并构建其超表达载体转化大豆毛状根,研究其对胞囊线虫病的抗性功能。结果表明,GmC2H2-2like编码410个氨基酸残基,分子质量为45.77 ku,等电点为8.73,含47个磷酸化位点,蛋白二级结构含α-螺旋(25.61%)、无规则卷曲(57.32%)、延伸链(13.66%)和β-转角(3.14%)4种结构;亚细胞定位结果发现,其编码蛋白位于细胞核;分析超表达转GmC2H2-2like大豆毛状根发现,其根部组织的单位面积胞囊线虫数量显著低于空载体对照,表明GmC2H2-2like具有抑制胞囊线虫的功能。

关键词: 大豆, 胞囊线虫, GmC2H2-2like, 生物信息学, 亚细胞定位

Abstract:

In order to study the important role of zinc finger protein C2H2 in the stress response to soybean cyst nematode disease(SCN),lay an important foundation for further research on anti-soybean cyst nematode.The transcriptome data of SCN3 roots inoculated with Dongnong L-10(SCN-resistant)and Heinong 37(SCN-susceptible)were utilized in the present study,and the differentially expressed gene,GmC2H2-2like,was screened out.The localization of this gene was observed under the inverted fluorescence microscope.The gene was bioinformatically analyzed,and its overexpression vector was cloned and constructed to transform soybean hairy roots to study its resistance function against cyst nematode disease.The results showed that GmC2H2-2like encoded 410 amino acid residues,with a molecular weight of 45.77 ku,an isoelectric point of 8.73,and 47 phosphorylation sites,and the protein tertiary structure contained four structures,namely,α-helix(25.61%),random coil(57.32%),extended strand(13.66%),and β-turn(3.14%);the results of the subcellular localization revealed that the encoded protein was located in the nucleus;analysis of the overexpressed trans-GmC2H2-2like soybean hairy roots revealed that the number of cyst nematodes per unit area of the root tissue was significantly lower than that of the empty vector control,suggesting that GmC2H2-2like has the function of inhibiting cyst nematodes.

Key words: Soybean, Cyst nematode, GmC2H2-2like, Bioinformatics, Subcellular localisation

引用本文

曲硕, 刘芳, 宋庚晨, 胡世豪, 房瑶瑶, 赵雪, 韩英鹏. 大豆抗胞囊线虫锌指蛋白基因GmC2H2-2like克隆与初步功能分析[J]. 华北农学报, 2024, 39(3): 195-207. doi: 10.7668/hbnxb.20194623.

QU Shuo, LIU Fang, SONG Gengchen, HU Shihao, FANG Yaoyao, ZHAO Xue, HAN Yingpeng. Soybean Cyst Nematode Resistant Zinc Finger Protein Gene GmC2H2-2like Cloning and Preliminary Functional Analysis[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(3): 195-207. doi: 10.7668/hbnxb.20194623.