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Creation and physiological analysis of an e1-as gene mutant in soybean

HE Hong-Li1,ZHANG Yu-Han1,YANG Jing2,CHENG Yun-Qing1,ZHAO Yang1,LI Xing-Nuo1,SI Hong-Liang3,ZHANG Xing-Zheng1,*,YANG Xiang-Dong2,*   

  1. 1 Jilin Normal University, Siping 136000, Jilin, China; 2 Jilin Academy of Agricultural Sciences, Changchun 130000, Jilin, China; 3 Boda College, Jilin Normal University, Siping 136000, Jilin, China
  • Received:2024-09-24 Revised:2025-04-25 Accepted:2025-04-25 Published:2025-05-13
  • Supported by:
    This study was supported by the National Natural Science Foundation of China (32101649) and the Outstanding Talents Team Project of Department of Science and Technology of Jilin Province (20240601063RC).

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Abstract: Glycine max is a photoperiod-sensitive plant, and E1 is a key gene regulating flowering time in soybean. As a core component of the photoperiodic flowering pathway, E1 plays a critical role in controlling the timing of floral transition. In this study, we constructed a CRISPR/Cas9-based genome editing vector targeting the E1 gene and introduced it into the soybean cultivar Williams 82. Gene editing was successfully detected in the E1 gene of T1 generation plants, and T2 generation plants were subsequently developed. Sequencing analysis revealed a single base insertion at target site 1 in the E1 gene, resulting in a premature stop codon and truncated protein. Phenotypic analysis showed that the flowering time of the T3 generation was approximately (10±2) days earlier than that of the wild type. In addition, chlorophyll content in the newly emerged trifoliate leaves of the mutant was (6±2) μg g?1 lower than in the wild type, while in fully expanded leaves, it was (10±4) μg g?1 lower. The pollen germination rate in the mutant was 8%±1% lower compared to the wild type. Furthermore, mutant pollen tube lengths measured at 0.25, 0.50, 1.00, and 2.00 h were (33.96±5.00), (74.14±5.00), (142.86±5.00), (183.50±5.00) μm, respectively, whereas the corresponding values in the wild type were (46.08±5.00), (118.89±5.00), (228.35±5.00), and (307.72±5.00) μm. Transcriptome analysis of the e1-as-79 mutant and the wild type identified 3,615 differentially expressed genes, including key flowering regulators FT2a and FT5a, which likely contribute to the early flowering phenotype observed in the mutant.

Key words: soybean, E1 gene, gene editing, mutant, transcriptome

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