Xu Jiao, Zheng Wei, Ou Xiaohong, He Hua, Yang Changgui, Guo Lanping, Xiao Chenghong, Jiang Weike, Shu Guoping, Zhou Tao
Institute of Traditional Chinese & Ethnic Medicine Resources, Guizhou University of Traditional Chinese Medicine, Guiyang, China.
Guizhou Key Laboratory for Germplasm Innovation and Resource-Efficient Utilization of Dao-di Herbs, Guizhou, Guiyang, China.
BMC Plant Biol. 2025 Aug 20;25(1):1103. doi: 10.1186/s12870-025-06972-2.
, a member of the Caryophyllaceae family, is widely used in traditional Chinese medicine due to its bioactive cyclic peptides (CPs) with immunomodulatory functions. Caryophyllaceae- like CPs, one of the largest types plant-derived CPs, typically consist of 5–12 amino acids and are derived from ribosomally synthesized peptide precursors. The diversity of CPs arises from variations in their core peptide sequences. However, the precursor genes responsible for Caryophyllaceae-like CPs biosynthesis in remain largely uncharacterized.
In this study, barcoding PCR combined with high-throughput sequencing was used to efficiently genotype precursor genes encoding CPs in . This approach enabled the identification of known and novel precursor genes, including , , and . The core peptide regions showed high variability, while the leader and follower regions were relatively conserved, with a few nucleotide mutations. Tissue-specific expression analysis revealed that was predominantly expressed in the phloem and fibrous roots, while was specifically expressed in the xylem. exhibited low expression level and was mainly detected in the phloem and stem. Moreover, the expression of these precursor genes was responsive to abscisic acid and nitrogen stress. RNA in situ hybridization revealed that transcripts were primarily localized in the xylem and phellem of the roots. Transient co-expression in indicated that is involved in the biosynthesis of Pseudostellarin E (PE).
Barcoding PCR combined with high-throughput sequencing provides an effective strategy for investigating CP precursor genes, including those with low expression. The results reveal conserved features in CP precursor genes and highlight a previously unrecognized mechanism contributing to CP diversity. The gene was identified as the precursor gene of PE, which accumulates mainly in the xylem of roots. may be a precursor gene for a novel CP.
The online version contains supplementary material available at 10.1186/s12870-025-06972-2.
石竹科植物，因其具有免疫调节功能的生物活性环肽（CPs）而在传统中药中广泛应用。石竹科样CPs是植物来源的最大类型CPs之一，通常由5 - 12个氨基酸组成，来源于核糖体合成的肽前体。CPs的多样性源于其核心肽序列的变异。然而，负责石竹科样CPs生物合成的前体基因在很大程度上仍未被表征。
在本研究中，条形码PCR结合高通量测序被用于高效地对编码石竹科植物CPs的前体基因进行基因分型。该方法能够鉴定已知和新的前体基因，包括、、和。核心肽区域显示出高度变异性，而前导和尾随区域相对保守，有一些核苷酸突变。组织特异性表达分析表明，主要在韧皮部和须根中表达，而特异性地在木质部中表达。表达水平较低，主要在韧皮部和茎中检测到。此外，这些前体基因的表达对脱落酸和氮胁迫有响应。RNA原位杂交显示，转录本主要定位于根的木质部和木栓。在中的瞬时共表达表明，参与了假繁缕素E（PE）的生物合成。
条形码PCR结合高通量测序为研究CP前体基因提供了一种有效的策略，包括那些低表达的基因。结果揭示了CP前体基因的保守特征，并突出了一种以前未被认识的导致CP多样性的机制。基因被鉴定为PE的前体基因，PE主要积累在石竹科植物根的木质部中。可能是一种新型CP的前体基因。
在线版本包含可在10.1186/s12870 - 025 - 06972 - 2获取的补充材料。
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