小麦转录因子单克隆酵母单/双杂文库构建与验证

doi:10.1016/j.jia.2026.07.037

Journal of Integrative Agriculture ›› 2026, Vol. 25 ›› Issue (10): 4378-4382.DOI: 10.1016/j.jia.2026.07.037

小麦转录因子单克隆酵母单/双杂文库构建与验证

收稿日期:2026-04-20 修回日期:2026-07-30 接受日期:2026-07-03 出版日期:2026-10-20 发布日期:2026-09-21

Construction and validation of a monoclonal yeast transcription factor library for high-throughput interaction screening in wheat

Kangzhen Ren^1*, Yalin Wang^1*, Yutao Cui^1*, Jinlan Ni¹, Chuangyi Yang¹, Weihang Sun¹, Chenfei Jia¹, Tingzhi Yang¹, Xinru Lü¹, Huimin Qin¹, Xiaohan Xie¹, Xianghua Meng¹, Wenjiao Zhang¹, Fulin Yang¹, Xuehuan Dai¹, Jianbin Zeng¹, Wenxing Liu¹, Faji Li^{3, 5}, Yuanfeng Hao^2#, Wujun Ma^{1, 4#}, Dengan Xu^{1, 3#}

¹ College of Agronomy, Qingdao Agricultural University, Qingdao 266109, China
² Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
³ Open Research Projects of the State Key Laboratory of Wheat Breeding, Jinan 250000, China
⁴ Australia-China Joint Centre for Wheat Improvement, State Agriculture Biotechnology Centre, Murdoch University, Perth, WA, 6150, Australia
⁵ Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250000, China

Received:2026-04-20 Revised:2026-07-30 Accepted:2026-07-03 Online:2026-10-20 Published:2026-09-21
About author:#Correspondence Dengan Xu, E-mail: xudengan@qau.edu.cn; Wujun Ma, E-mail: wujun.ma@qau.edu.cn; Yuanfeng Hao, E-mail: haoyuanfeng@caas.cn * These authors contributed equally to this study.
Supported by:
This work was funded by the National Natural Science Foundation of China Regional Innovation and Development Joint Fund Project (U22A20457), the Open Research Projects of the State Key Laboratory of Wheat Improvement, China, the National Natural Science Foundation of China (32101733), the Shandong Provincial Natural Science Foundation, China (ZR2021QC164), and the Shandong Agricultural Seeds Engineering Project, China (2023LZGC009).

摘要/Abstract

摘要：

转录因子（TFs）能够结合顺式作用元件、形成蛋白复合物，进而调控基因表达。小麦为多倍体物种，基因组庞大，且转录因子表达水平存在显著差异，基于总mRNA构建的传统酵母双杂交（Y2H）、酵母单杂交（Y1H）文库筛选效率有限。本研究整合PlantTFDB数据库与中国春参考基因组（IWGSC v1.1）注释信息，共鉴定到6230个小麦候选转录因子，并筛选其中5763个转录因子（TPM≥0.1）用于文库构建。针对各转录因子设计逐一特异性引物，从26种不同组织的cDNA中成功克隆得到3142个转录因子，成功构建靶向单克隆酵母转录因子文库，覆盖度约55%。以春化基因VRN1、VRN2和VRN3作为诱饵开展文库筛选，分别获得97、237、173个互作转录因子；其中27个转录因子可同时与上述3个春化开花基因发生互作。利用荧光素酶互补实验对代表性互作关系进行验证，并结合已发表RNA-Seq数据解析了3个春化开花基因在表达模式与调控网络上的差异。该富集转录因子文库可同时适用于酵母单杂交与酵母双杂交体系，为系统研究小麦转录因子调控网络提供了有效工具。

Kangzhen Ren, Yalin Wang, Yutao Cui, Jinlan Ni, Chuangyi Yang, Weihang Sun, Chenfei Jia, Tingzhi Yang, Xinru Lü, Huimin Qin, Xiaohan Xie, Xianghua Meng, Wenjiao Zhang, Fulin Yang, Xuehuan Dai, Jianbin Zeng, Wenxing Liu, Faji Li, Yuanfeng Hao, Wujun Ma, Dengan Xu. 小麦转录因子单克隆酵母单/双杂文库构建与验证[J]. Journal of Integrative Agriculture, 2026, 25(10): 4378-4382.

Kangzhen Ren, Yalin Wang, Yutao Cui, Jinlan Ni, Chuangyi Yang, Weihang Sun, Chenfei Jia, Tingzhi Yang, Xinru Lü, Huimin Qin, Xiaohan Xie, Xianghua Meng, Wenjiao Zhang, Fulin Yang, Xuehuan Dai, Jianbin Zeng, Wenxing Liu, Faji Li, Yuanfeng Hao, Wujun Ma, Dengan Xu. Construction and validation of a monoclonal yeast transcription factor library for high-throughput interaction screening in wheat[J]. Journal of Integrative Agriculture, 2026, 25(10): 4378-4382.

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小麦转录因子单克隆酵母单/双杂文库构建与验证

Construction and validation of a monoclonal yeast transcription factor library for high-throughput interaction screening in wheat

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