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Background <p><i>Arctium lappa</i> L<i>.</i> is a traditional Chinese medicine, and arctiin is the main active ingredient of <i>A. lappa</i>, which is a lignan. In the biosynthetic pathway of arctiin, dirigent proteins (<i>DIRs</i>) play a critical role in mediating the stereoisomerization of coniferyl alcohol in its phenoxy radical coupling reaction to produce optically active ( +) or ( −)-pinoresinol. However, the <i>DIRs</i> in <i>A. lappa</i> (<i>AlDIR</i>s) have not been studied in detail. To identify <i>AlDIR</i> genes and clarify their roles in arctiin biosynthesis, this study analyzed <i>AlDIR</i>s, verified the catalytic activity of the <i>AlDIR</i>1, and examined the stress-resistant effects of <i>AlDIR</i>s.</p> Results <p>For the first time, 17 <i>AlDIR</i>s were identified in <i>A. lappa.</i> The database showed that <i>AlDIR</i>s were located on chromosomes 0, 2, 3, 6, 8, 9, and 11. Gene duplication analysis revealed five pairs of fragment duplication events in the <i>AlDIR</i> family. Since <i>AlDIR</i>1 belongs to the DIR-a subfamily, which is known to catalyze the formation of ( ±)-pinoresinol, its activity was further investigated. By LC–MS, it was confirmed that recombinant AlDIR1 proteins convert coniferyl alcohol into ( +)-pinoresinol. Significant changes in <i>AlDIR</i> were observed in both roots and leaves of <i>A. lappa</i> after treatment by abscisic acid (ABA), salicylic acid (SA), NaCl, polyethylene glycol (PEG) hormonal stresses, and the role of <i>AlDIR</i>s in abiotic stress resistance has been demonstrated.</p> Conclusion <p>The results of this study reveal the molecular and stress resistance properties of the DIR family in <i>A. lappa</i>, providing a basis for further research on the DIR gene family and the promotion of arctiin biosynthesis.</p>

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Identification of the Dirigent Genes in Arctium lappa and their Relevance to the Stress Response

  • 期刊论文

  • 发表日期：

    2025年7月30日

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  • Mingze Gao,
  • Ying Yu,
  • Jianing Xue,
  • Yilin Han,
  • Rong Zhao

摘要

Background

Arctium lappa L. is a traditional Chinese medicine, and arctiin is the main active ingredient of A. lappa, which is a lignan. In the biosynthetic pathway of arctiin, dirigent proteins (DIRs) play a critical role in mediating the stereoisomerization of coniferyl alcohol in its phenoxy radical coupling reaction to produce optically active ( +) or ( −)-pinoresinol. However, the DIRs in A. lappa (AlDIRs) have not been studied in detail. To identify AlDIR genes and clarify their roles in arctiin biosynthesis, this study analyzed AlDIRs, verified the catalytic activity of the AlDIR1, and examined the stress-resistant effects of AlDIRs.

Results

For the first time, 17 AlDIRs were identified in A. lappa. The database showed that AlDIRs were located on chromosomes 0, 2, 3, 6, 8, 9, and 11. Gene duplication analysis revealed five pairs of fragment duplication events in the AlDIR family. Since AlDIR1 belongs to the DIR-a subfamily, which is known to catalyze the formation of ( ±)-pinoresinol, its activity was further investigated. By LC–MS, it was confirmed that recombinant AlDIR1 proteins convert coniferyl alcohol into ( +)-pinoresinol. Significant changes in AlDIR were observed in both roots and leaves of A. lappa after treatment by abscisic acid (ABA), salicylic acid (SA), NaCl, polyethylene glycol (PEG) hormonal stresses, and the role of AlDIRs in abiotic stress resistance has been demonstrated.

Conclusion

The results of this study reveal the molecular and stress resistance properties of the DIR family in A. lappa, providing a basis for further research on the DIR gene family and the promotion of arctiin biosynthesis.