摘要
后期促进复合物(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(H_(2)O_(2))和超氧阴离子(O_(2)^(-))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。
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 up-regulated 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(H_(2)O_(2))and superoxide anion(O_(2)^(-))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.
作者
胡城祯
高维东
孔斌雪
王建飞
车卓
杨德龙
陈涛
Hu Cheng-Zhen;Gao Wei-Dong;Kong Bin-Xue;Wang Jian-Fei;Che Zhuo;Yang De-Long;Chen Tao(State Key Laboratory of Aridland Crop Science,Gansu Agricultural University,Lanzhou 730070,Gansu,China;College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,Gansu,China;Agronomy College,Gansu Agricultural University,Lanzhou 730070,Gansu,China)
出处
《作物学报》
北大核心
2026年第1期148-164,共17页
Acta Agronomica Sinica
基金
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)
甘肃省教育厅2025年研究生创新之星项目(2025CXZX-853)资助。
