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5-AzaC Facilitates Somatic Embryogenesis and Germination Across Two Embryogenic Lines in Larix olgensis
 
 
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Volume 14
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10.3390/plants14203127
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

A Cryopreservation and Regeneration Protocol for Embryogenic Callus of Larix olgensis

by
Chen Wang
,
Wenna Zhao
,
Yu Liu
,
Hao Dong
,
Yajing Ning
,
Chengpeng Cui
,
Hanguo Zhang
,
Meng Li
* and
Shujuan Li
*
State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China
*
Authors to whom correspondence should be addressed.
Plants 2025, 14(20), 3127; https://doi.org/10.3390/plants14203127
Submission received: 17 September 2025 / Revised: 5 October 2025 / Accepted: 9 October 2025 / Published: 10 October 2025
(This article belongs to the Special Issue Sexual and Asexual Reproduction in Forest Plants—2nd Edition)
Versions Notes

Abstract

Larix olgensis is a valuable timber species in northern China, typically propagated through somatic embryogenesis (SE). However, long-term subculture can lead to a loss of embryogenic potential. This study aimed to establish a simple and stable protocol for the cryopreservation and regeneration of L. olgensis embryogenic callus (EC) that preserves its SE potential and regenerative capacity. The slow-freezing method was employed for cryopreservation. A cryopreservation protocol for L. olgensis EC was developed by optimizing the preculture duration and conditions, cryoprotectant composition and thawing temperature. The results showed that optimal outcomes were achieved using a 24 h stepwise preculture on medium containing 0.2 and 0.4 mol∙L−1 sucrose, followed by cryoprotectant treatment with 0.4 mol∙L−1 sucrose, 2.5% (v/v) dimethyl sulfoxide (DMSO) and 10% polyethylene glycol 6000 (PEG6000), and thawing at 37 °C. EC cryopreserved using this protocol achieved a 100% recovery rate. Moreover, the cryopreserved recoverable EC successfully underwent SE, progressing through germination and rooting. Cryopreservation duration (storage duration in liquid nitrogen) did not affect cell viability and proliferation rate, confirming the protocol’s suitability for long-term cryopreservation of L. olgensis EC. This study provides a valuable reference for the cryopreservation and regeneration of L. olgensis EC, with potential applications for other coniferous species. It establishes a robust foundation for the large-scale propagation of conifers.
Keywords: preculture; cryoprotectant; thawing temperature; cell viability; proliferation rate; germplasm preservation preculture; cryoprotectant; thawing temperature; cell viability; proliferation rate; germplasm preservation

Share and Cite

MDPI and ACS Style

Wang, C.; Zhao, W.; Liu, Y.; Dong, H.; Ning, Y.; Cui, C.; Zhang, H.; Li, M.; Li, S. A Cryopreservation and Regeneration Protocol for Embryogenic Callus of Larix olgensis. Plants 2025, 14, 3127. https://doi.org/10.3390/plants14203127

AMA Style

Wang C, Zhao W, Liu Y, Dong H, Ning Y, Cui C, Zhang H, Li M, Li S. A Cryopreservation and Regeneration Protocol for Embryogenic Callus of Larix olgensis. Plants. 2025; 14(20):3127. https://doi.org/10.3390/plants14203127

Chicago/Turabian Style

Wang, Chen, Wenna Zhao, Yu Liu, Hao Dong, Yajing Ning, Chengpeng Cui, Hanguo Zhang, Meng Li, and Shujuan Li. 2025. "A Cryopreservation and Regeneration Protocol for Embryogenic Callus of Larix olgensis" Plants 14, no. 20: 3127. https://doi.org/10.3390/plants14203127

APA Style

Wang, C., Zhao, W., Liu, Y., Dong, H., Ning, Y., Cui, C., Zhang, H., Li, M., & Li, S. (2025). A Cryopreservation and Regeneration Protocol for Embryogenic Callus of Larix olgensis. Plants, 14(20), 3127. https://doi.org/10.3390/plants14203127

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