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Metabolites
Volume 16
Issue 6
10.3390/metabo16060374
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Transcriptomic and Metabolomic Insights into the Enhanced Quality of Anoectochilus roxburghii Seedlings in Sugar-Free Versus Conventional Tissue Culture Systems

by
Xiangtao Chen
1,†,
Fangfang Chen
2,†,
Chuanzhi Kang
3,†,
Tongwei Lin
1,
Hongyang Wang
3,
Yiheng Wang
3,
Dehua Wu
3,
Wanying Duan
3,
Zekun Zhang
3,* and
Chengcai Zhang
3,*
1
Lishui Jianlan Biotechnology Co., Ltd., Lishui 323500, China
2
Hangzhou Mumu Biotechnology Co., Ltd., Hangzhou 311300, China
3
State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Metabolites 2026, 16(6), 374; https://doi.org/10.3390/metabo16060374
Submission received: 29 April 2026 / Revised: 20 May 2026 / Accepted: 25 May 2026 / Published: 29 May 2026
(This article belongs to the Section Plant Metabolism)
Review Reports Versions Notes

Abstract

Background/Objective: Anoectochilus roxburghii, a high-value medicinal orchid, faces significant challenges in quality standardization during large-scale tissue culture due to a lack of understanding of the underlying molecular mechanisms. This study aimed to compare “Jianlan No.2” plantlets cultured under a conventional tissue culture system (CK) and a sugar-free tissue culture system (TD), to elucidate the phenotypic and molecular basis for quality improvement. Methods: A systematic comparison was conducted. Phenotypic traits of plantlets from both systems were measured. Integrated transcriptomic (RNA sequencing) and untargeted metabolomic analyses were employed to identify the molecular differences at the gene expression and metabolite accumulation levels. Results: TD-grown seedlings exhibited significantly superior growth characteristics, including greater plant height, higher rooting rate, and improved transplant survival. Transcriptomic analysis identified 416 differentially expressed genes (DEGs) (44 upregulated, 372 downregulated in TD), which were significantly enriched in pathways related to cell wall organization, apoplast, and photosynthesis. Sixteen key genes were pinpointed as closely associated with seedling growth and metabolic regulation. Metabolomic profiling revealed 502 differentially accumulated metabolites (DAMs), with significant perturbations primarily in phenylpropanoid biosynthesis and terpenoid metabolism. Conclusions: The sugar-free tissue culture system enhances A. roxburghii seedling quality by coordinately modulating photosynthetic capacity, carbon metabolism, and the biosynthesis of key secondary metabolites. These findings provide a crucial molecular foundation for optimizing tissue culture protocols and advancing the standardized, high-quality cultivation of this valuable medicinal plant.
Keywords: Anoectochilus roxburghii; cultivation methods; metabolome; sugar-free tissue culture; transcriptome Anoectochilus roxburghii; cultivation methods; metabolome; sugar-free tissue culture; transcriptome

Share and Cite

MDPI and ACS Style

Chen, X.; Chen, F.; Kang, C.; Lin, T.; Wang, H.; Wang, Y.; Wu, D.; Duan, W.; Zhang, Z.; Zhang, C. Transcriptomic and Metabolomic Insights into the Enhanced Quality of Anoectochilus roxburghii Seedlings in Sugar-Free Versus Conventional Tissue Culture Systems. Metabolites 2026, 16, 374. https://doi.org/10.3390/metabo16060374

AMA Style

Chen X, Chen F, Kang C, Lin T, Wang H, Wang Y, Wu D, Duan W, Zhang Z, Zhang C. Transcriptomic and Metabolomic Insights into the Enhanced Quality of Anoectochilus roxburghii Seedlings in Sugar-Free Versus Conventional Tissue Culture Systems. Metabolites. 2026; 16(6):374. https://doi.org/10.3390/metabo16060374

Chicago/Turabian Style

Chen, Xiangtao, Fangfang Chen, Chuanzhi Kang, Tongwei Lin, Hongyang Wang, Yiheng Wang, Dehua Wu, Wanying Duan, Zekun Zhang, and Chengcai Zhang. 2026. "Transcriptomic and Metabolomic Insights into the Enhanced Quality of Anoectochilus roxburghii Seedlings in Sugar-Free Versus Conventional Tissue Culture Systems" Metabolites 16, no. 6: 374. https://doi.org/10.3390/metabo16060374

APA Style

Chen, X., Chen, F., Kang, C., Lin, T., Wang, H., Wang, Y., Wu, D., Duan, W., Zhang, Z., & Zhang, C. (2026). Transcriptomic and Metabolomic Insights into the Enhanced Quality of Anoectochilus roxburghii Seedlings in Sugar-Free Versus Conventional Tissue Culture Systems. Metabolites, 16(6), 374. https://doi.org/10.3390/metabo16060374

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