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小麦TaAPC11基因家族鉴定及TaAPC11-5B参与干旱胁迫的生物学功能研究

胡城祯1,2，高维东1,2，孔斌雪1,3，王建飞1,2，车卓1，杨德龙1,2,*，陈涛1,2,*   

  1. Hu Cheng-Zhen1,2,Gao Wei-Dong1,2,Kong Bing-Xue1,3,Wang Jian-Fei1,2,Che Zhuo1,Yang De-Long1,2,*,Chen Tao1,2,*
  • 收稿日期:2025-03-13 修回日期:2025-08-13 接受日期:2025-08-13 网络出版日期:2025-08-22
  • 基金资助:
    本研究由甘肃省农业农村厅种业攻关项目(GYGG-2024-2), 甘肃省科技厅基础研究创新群体项目(24JRRA633), 甘肃省科技重大专项(22ZD6NA009), 国家自然科学基金项目(32360518, 32260520, 32160487), 甘肃省高校科研创新平台重点培育项目(2024CXPT-01), 中央引导地方科技发展资金项目(23ZYQA0322), 甘肃省高等学校产业支撑计划项目(2022CYZC-44), 甘肃省教育厅青年博士支持项目(2024QB-062), 省部共建干旱生境作物学国家重点实验室开放基金项目(GSCS-2023-07), 甘肃省重点研发计划(25YFWA020)和甘肃省教育厅2025年研究生创新之星项目(2025CXZX-853)资助。

Genome-wide identification of the TaAPC11 gene family in wheat and functional characterization of TaAPC11-5B in drought stress responses

Hu Cheng-Zhen1,2,Gao Wei-Dong1,2,Kong Bing-Xue1,3,Wang Jian-Fei1,2,Che Zhuo1,Yang De-Long1,2,*,Chen Tao1,2,*   

  1. 1 State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Lanzhou 730070, Gansu, China; 2 College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, Gansu, China; 3 College of Agricultural, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2025-03-13 Revised:2025-08-13 Accepted:2025-08-13 Published online:2025-08-22
  • Supported by:
    This study was supported by Breakthrough Project in Seed Industry of Gansu Province (GYGG-2024-2), Innovative Research Group Project of Gansu Province (24JRRA633), the Key Science and Technology Special Project of Gansu Province (22ZD6NA009), the National Natural Science Foundation of China (32360518, 32260520, 32160487), Key Cultivation Project of University Research and Innovation Platform of Gansu Province (2024CXPT-01), the Development Fund Project of National Guiding Local Science and Technology (23ZYQA0322), the Industrial Support Plan of Colleges and Universities in Gansu Province (2022CYZC-44), Gansu Provincial Education Department Young Doctor Support Project (2024QB-062), Open Fund for the State Key Laboratory of Crop Science in Arid Habitat Jointly Established by the Ministry of Provinces and China (GSCS-2023-07), Key Research and Development Program of Gansu Province (25YFWA020), and the Graduate Innovation Star Project 2025 by Gansu Provincial Education Department, China (2025CXZX-853).

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摘要：

后期促进复合物(The anaphase-promoting complex/cyclosome, APC/C)是一种由多亚基组成的cullin-RING型E3泛素连接酶，其中复合物亚基APC11含有RING结构域，可能通过泛素化特定的靶蛋白调控细胞周期，进而参与植物的非生物胁迫。本研究采用生物信息学方法对小麦TaAPC11家族成员进行全基因组鉴定，并重点研究了TaAPC11-5B基因参与干旱胁迫调控的生物学功能。结果表明，小麦基因组中共鉴定到23个TaAPC11家族成员，可分为3个亚族，同一亚族成员具有相似的基因结构和Motif。共线性分析表明，片段复制促进TaAPC11基因家族的扩张，且在进化过程中受到纯化选择。顺式作用元件分析表明，TaAPC11基因家族成员的启动子区存在大量非生物胁迫响应元件。实时荧光定量(qRT-PCR)分析表明，TaAPC11-5B/6A2/4D1/3B2基因的转录表达均受到PEG-6000和ABA的显著诱导，其中TaAPC11-5B在PEG-6000处理后上调倍数最高。亚细胞定位表明，TaAPC11-5B定位于细胞核与细胞质中。对野生型(WT)和TaAPC11转基因水稻株系(TaAPC11-5B-OE)进行干旱胁迫处理，分析其抗旱表型发现，TaAPC11-5B-OE的存活率显著高于WT，株高显著增加，叶片卷曲程度减轻；生理指标测定发现，TaAPC11-5B-OE的相对电导率和丙二醛含量显著低于WT，脯氨酸含量显著高于WT。进一步通过DAB (3,3'-二氨基联苯胺)、NBT (氮蓝四唑)染色及抗氧化酶活性分析发现，PEG-6000处理后TaAPC11-5B-OE叶片的过氧化氢(H2O2)和超氧阴离子(O2?)含量低于WT，过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)的活性均显著高于WT。本研究为深入解析小麦TaAPC11-5B基因响应干旱胁迫的生物学功能提供了理论依据和材料基础。

关键词: 小麦, 干旱胁迫, 基因家族, TaAPC11-5B, 表达模式, 耐旱功能研究

Abstract:

The anaphase-promoting complex/cyclosome (APC/C), a multi-subunit cullin-RING-type E3 ubiquitin ligase, regulates cell cycle progression by ubiquitinating specific target proteins via its RING domain-containing subunit, APC11, thereby contributing to plant responses to abiotic stress. In this study, we performed a genome-wide identification of the TaAPC11 gene family in wheat using bioinformatics approaches, with a particular focus on elucidating the biological function of TaAPC11-5B in drought stress regulation. A total of 23 TaAPC11 members were identified in the wheat genome and classified into three subfamilies, all exhibiting conserved gene structures and motifs. Synteny analysis revealed that segmental duplications, driven by purifying selection, contributed to the expansion of the TaAPC11 family. Cis-acting element analysis indicated an abundance of abiotic stress-responsive elements in the promoters of TaAPC11 genes. Quantitative reverse transcription PCR (qRT-PCR) analysis showed that transcripts of TaAPC11-5B, 6A2, 4D1, and 3B2 were significantly upregulated by PEG-6000 and ABA treatments, with TaAPC11-5B exhibiting the strongest response under PEG-6000-induced drought stress. Comparative analysis between wild-type (WT) and TaAPC11-5B-overexpressing rice lines (TaAPC11-5B-OE) under PEG-6000 treatment demonstrated that the overexpression lines had significantly higher survival rates, increased plant height, and a reduced leaf rolling index compared to WT plants. Physiological assays further revealed that TaAPC11-5B-OE plants exhibited lower relative electrolyte leakage and malondialdehyde (MDA) content, but higher proline accumulation under drought conditions. Moreover, DAB (3,3'-diaminobenzidine) and NBT (nitroblue tetrazolium) staining, along with antioxidant enzyme activity assays, showed that TaAPC11-5B-OE plants accumulated less hydrogen peroxide (H2O2) and superoxide anion (O2?) while displaying enhanced activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD). Collectively, these results suggest that TaAPC11-5B enhances drought tolerance by modulating reactive oxygen species (ROS) scavenging. This study provides a theoretical basis and genetic resource for further understanding the drought-responsive mechanisms mediated by TaAPC11-5B in wheat.

Key words: wheat, drought stress, gene family, TaAPC11-5B, expression pattern, drought-tolerance validation

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