[1]朱小雨,谭庆,姚瑶,等.小麦4-羟基苯丙酮酸双氧化酶(TaHPPD)基因克隆与功能鉴定[J].江苏农业学报,2026,42(01):8-20.[doi:doi:10.3969/j.issn.1000-4440.2026.01.002]
　ZHU Xiaoyu,TAN Qing,YAO Yao,et al.Cloning and functional identification of wheat 4-hydroxyphenylpyruvate dioxygenase (TaHPPD) gene[J].,2026,42(01):8-20.[doi:doi:10.3969/j.issn.1000-4440.2026.01.002]
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小麦4-羟基苯丙酮酸双氧化酶(TaHPPD)基因克隆与功能鉴定()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
42
期数:
2026年01期
页码:
8-20
栏目:
遗传育种·生理生化
出版日期:
2026-01-31

文章信息/Info

Title:
Cloning and functional identification of wheat 4-hydroxyphenylpyruvate dioxygenase (TaHPPD) gene
作者:
朱小雨1; 谭庆2; 姚瑶3; 杨郁文3; 张保龙3
(1.扬州大学生物科学与技术学院,江苏扬州225009；2.海南大学热带农林学院,海南海口570228；3.江苏省农业科学院种质资源与生物技术研究所,江苏南京210014)
Author(s):
ZHU Xiaoyu1; TAN Qing2; YAO Yao3; YANG Yuwen3; ZHANG Baolong3
(1.College of Bioscience and Biotechnology, Yangzhou University, Yangzhou 225009, China;2.School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China;3.Institute of Germplasm Resources and Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
小麦; 4-羟基苯丙酮酸双氧化酶; 硝磺草酮; 除草剂抗性
Keywords:
wheat; 4-hydroxyphenylpyruvate dioxygenase; mesotrione; herbicide resistance
分类号:
S512.1
DOI:
doi:10.3969/j.issn.1000-4440.2026.01.002
文献标志码:
A
摘要:
为明确小麦除草剂靶基因HPPD的特征与功能,本研究克隆了小麦HPPD基因并对其进行序列分析、表达特征分析、亚细胞定位、原核表达、蛋白酶活性及功能分析等。结果表明,小麦有3个HPPD等位基因,分别位于6A、6B和6D染色体,其中TaHPPD-A与TaHPPD-B亲缘关系较近。TaHPPD启动子含有大量光响应元件和逆境响应元件。光照处理12 h,TaHPPD-A及TaHPPD-D的表达量分别比暗处理结束时（对照）下降80.9%和63.3%,光照处理24 h,TaHPPD-A、TaHPPD-B及TaHPPD-D表达量分别比CK下降88.1%、40.4%和63.6%。TaHPPD的表达受赤霉病、条锈病及干旱等逆境胁迫诱导。亚细胞定位分析结果表明TaHPPD-A蛋白位于细胞质与叶绿体,而TaHPPD-D蛋白仅定位于叶绿体。TaHPPD蛋白在较高浓度硝磺草酮显色液中仍能显色,但蛋白质表达量显著高于玉米HPPD蛋白和水稻HPPD蛋白,但小麦HPPD酶活性显著低于水稻HPPD酶活性和玉米HPPD酶活性。相对于野生型水稻中花11，3个相对表达量最高的TaHPPD过表达植株(TaHPPD-A-OE-18、TaHPPD-B-OE-7和TaHPPD-D-OE-7)对HPPD抑制剂类除草剂硝磺草酮的抗性指数分别为13.92、15.66、7.63。本研究结果可为抗HPPD抑制剂类除草剂小麦种质的创制提供基础与依据。
Abstract:
In order to clarify the characteristics and functions of the wheat herbicide target gene HPPD, the wheat HPPD genes were cloned, and sequence analysis, expression feature analysis, subcellular localization, prokaryotic expression, enzymatic activity assays and functional analysis of the wheat HPPD genes were conducted in this study. The results showed that there were three HPPD alleles in wheat, which were located on chromosomes 6A, 6B and 6D, respectively, and TaHPPD-A was closely related to TaHPPD-B. The TaHPPD promoter contained a large number of light response elements and stress response elements. After 12 h of light treatment, the expression levels of TaHPPD-A and TaHPPD-D decreased by 80.9% and 63.3%, respectively, compared with the end of dark treatment (control). After 24 h of light treatment, the expression levels of TaHPPD-A, TaHPPD-B and TaHPPD-D decreased by 88.1%, 40.4% and 63.6%, respectively, compared with the control. The expression of TaHPPD was induced by stress conditions such as Fusarium head blight, stripe rust and drought. Subcellular localization analysis showed that TaHPPD-A protein was located in cytoplasm and chloroplast, while TaHPPD-D protein was only located in chloroplast. TaHPPD protein retained chromogenic activity even in a high-concentration mesotrione chromogenic solution. However, its protein expression level was significantly higher than that of corn HPPD protein and rice HPPD protein, while HPPD enzyme activity of wheat was significantly lower than that of rice and corn. Compared with the wild type rice Zhonghua11, the resistance indices of the three TaHPPD overexpression plants (TaHPPD-A-OE-18, TaHPPD-B-OE-7 and TaHPPD-D-OE-7) with the highest relative expression level were 13.92, 15.66 and 7.63, respectively. The results of this study provide a basis for the development of wheat germplasm resistant to HPPD inhibitor herbicides.

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备注/Memo

备注/Memo:
收稿日期：2025-03-13基金项目：江苏省农业科技自主创新基金项目[CX(21)1002]作者简介：朱小雨(1999-),女,安徽蚌埠人,硕士研究生,主要从事作物抗除草剂育种研究。(E-mail)1247239590@qq.com通讯作者：杨郁文,(E-mail)13814001979@139.com
更新日期/Last Update: 2026-02-09