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Volume 15
Issue 1
10.3390/plants15010018
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Enhancing Cryopreservation Efficiency in Populus davidiana × P. tremuloides Shoot Tips: Optimization of Vitrification Protocols and Mechanistic Insights into Flavonoid-Mediated Stress Adaptation

by
Panke Yang
1,†,
Zelin Li
1,†,
Yu Qi
1,
Yuandong Ma
1,
Chunming Li
1,
Maolan Liu
1,
Wenjun Ma
1,
Hui Bai
2,* and
Huanzhen Liu
1,*
1
College of Forestry, Northeast Forestry University, Harbin 150040, China
2
Heilongjiang Provincial Forestry Research Institute, Harbin 150040, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work and share first authorship.
Plants 2026, 15(1), 18; https://doi.org/10.3390/plants15010018 (registering DOI)
Submission received: 20 November 2025 / Revised: 15 December 2025 / Accepted: 18 December 2025 / Published: 20 December 2025
(This article belongs to the Section Plant Physiology and Metabolism)
Versions Notes

Abstract

Cryopreservation is vital for conserving the elite germplasm of the hybrid poplar Populus davidiana × P. tremuloides, which is difficult to propagate conventionally. This study established optimized vitrification and encapsulation–vitrification protocols, achieving high regeneration rates of 85.91% and 79.70%, respectively, with confirmed genetic stability. The process induced oxidative stress, altering markers (MDA, H2O2) and antioxidant enzyme activities (SOD, POD, CAT). Integrated transcriptomic and metabolomic analysis of key steps—preculture/loading (DLA) and osmotic dehydration (DLB)—revealed extensive stress-responsive reprogramming. A central finding was the robust activation of the flavonoid biosynthesis pathway during DLB, marked by upregulation of key genes (PAL, CHS) and accumulation of flavonols (e.g., quercetin). This response was linked to hormone signaling and antioxidant systems, forming a coordinated defense network. Our multi-omics findings demonstrate that successful cryopreservation relies on an adaptive response where flavonoid biosynthesis plays a critical role in conferring oxidative stress tolerance, providing a theoretical basis for improving woody plant cryopreservation.
Keywords: Populus davidiana × P. tremuloides; cryopreservation; vitrification; flavonoid biosynthesis; oxidative stress; multi-omics analysis Populus davidiana × P. tremuloides; cryopreservation; vitrification; flavonoid biosynthesis; oxidative stress; multi-omics analysis

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MDPI and ACS Style

Yang, P.; Li, Z.; Qi, Y.; Ma, Y.; Li, C.; Liu, M.; Ma, W.; Bai, H.; Liu, H. Enhancing Cryopreservation Efficiency in Populus davidiana × P. tremuloides Shoot Tips: Optimization of Vitrification Protocols and Mechanistic Insights into Flavonoid-Mediated Stress Adaptation. Plants 2026, 15, 18. https://doi.org/10.3390/plants15010018

AMA Style

Yang P, Li Z, Qi Y, Ma Y, Li C, Liu M, Ma W, Bai H, Liu H. Enhancing Cryopreservation Efficiency in Populus davidiana × P. tremuloides Shoot Tips: Optimization of Vitrification Protocols and Mechanistic Insights into Flavonoid-Mediated Stress Adaptation. Plants. 2026; 15(1):18. https://doi.org/10.3390/plants15010018

Chicago/Turabian Style

Yang, Panke, Zelin Li, Yu Qi, Yuandong Ma, Chunming Li, Maolan Liu, Wenjun Ma, Hui Bai, and Huanzhen Liu. 2026. "Enhancing Cryopreservation Efficiency in Populus davidiana × P. tremuloides Shoot Tips: Optimization of Vitrification Protocols and Mechanistic Insights into Flavonoid-Mediated Stress Adaptation" Plants 15, no. 1: 18. https://doi.org/10.3390/plants15010018

APA Style

Yang, P., Li, Z., Qi, Y., Ma, Y., Li, C., Liu, M., Ma, W., Bai, H., & Liu, H. (2026). Enhancing Cryopreservation Efficiency in Populus davidiana × P. tremuloides Shoot Tips: Optimization of Vitrification Protocols and Mechanistic Insights into Flavonoid-Mediated Stress Adaptation. Plants, 15(1), 18. https://doi.org/10.3390/plants15010018

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