Potential role of ice-binding protein in mitochondria-lipid and ATP mechanisms during freezing of plant callus

• Published in: Plant physiology and biochemistry 2024-09, Vol.214, p.108866, Article 108866
• Main Authors: Kim, Euihyun, Kwon, Gi-Sok, Choi, Sunmee, Kim, Soo-Yun, Heo, Kyeong Yeon, Kim, Young Soon, Kim, Cha Young, Kim, Soyoung, Jeong, Jae Cheol, Hwang, Jisub, Lee, Jun Hyuck, Lee, Jeong Hun, Moh, Sang Hyun
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - metabolism; Basidiomycota - metabolism; callus; cell culture; Cryopreservation; Cryopreservation - methods; Extracellular matrix; Freezing; Fungal Proteins - genetics; Fungal Proteins - metabolism; Ice-binding protein; Leucosporidium; Liliopsida; Lipid Metabolism; lipophilicity; Magnoliopsida; metabolism; mitochondria; Mitochondria - metabolism; Mitochondrial function; Plant callus; plant physiology; reactive oxygen species; Reactive Oxygen Species - metabolism; species; tissue culture; yeasts
• ISSN: 0981-9428; 1873-2690; 1873-2690
• DOI: 10.1016/j.plaphy.2024.108866

Plant calli, a perpetually undifferentiated cell culture, have defects in maintaining their genetic fidelity during prolonged tissue culture. Cryopreservation using ice-binding proteins (IBP) is a potential solution. Despite a few studies on cryopreservation using IBPs in plant calli, detailed insights into the intracellular metabolism during freezing, thawing, and re-induction remain sparse. This study investigated and employed IBP from polar yeast Leucosporidium sp. (LeIBP) in the cryopreservation process across diverse taxa, including gymnosperms, monocots, dicots, and woody plants. Molecular-level analyses encompassing reactive oxygen species levels, mitochondrial function, and ATP and lipophilic compounds content were conducted. The results across nine plant species revealed the effects of LeIBP on callus competency post-thawing, along with enhanced survival rates, reactive oxygen species reduction, and restored metabolic activities to the level of those of fresh calli. Moreover, species-specific survival optimization with LeIBP treatments and morphological assessments revealed intriguing extracellular matrix structural changes post-cryopreservation, suggesting a morphological strategy for maintaining the original cellular states and paracrine signaling. This study pioneered the comprehensive application of LeIBP in plant callus cryopreservation, alleviating cellular stress and enhancing competence. Therefore, our findings provide new insights into the identification of optimal LeIBP concentrations, confirmation of genetic conformity post-thawing, and the intracellular metabolic mechanisms of cryopreservation advancements in plant research, thereby addressing the challenges associated with long-term preservation and reducing labor-intensive cultivation processes. This study urges a shift towards molecular-level assessments in cryopreservation protocols for plant calli, advocating a deeper understanding of callus re-induction mechanisms and genetic fidelity post-thawing. [Display omitted] •LeIBP ensures genetic stability, preserving the integrity of diverse plant calluses.•Optimal LeIBP treatment enhances callus robustness during cryopreservation steps.•Reduced oxidative stress with LeIBP maintains intact cell competency in regeneration.•Post-thawing, LeIBP sustains mitochondrial activity, ATP, and FA for callus integrity.•LeIBP reinforces ECM integrity, ensuring successful callus recovery and cryopreservation.