Comprehensive molecular genetic characterization of the ICE1 gene in Arum korolkowii and functional insights into its encoded protein

Document Type : Research Paper

Authors

1 Zhangir Khan West Kazakhstan University, Kazakhstan

2 Department of Biotechnology, M. Auezov South Kazakhstan University, Shymkent 160000, Kazakhstan

3 Karakalpak State University named after Berdakh, Faculty of Biology, Nukus, 230112, Uzbekistan

4 3South Kazakhstan Medical Academy, Shymkent 160000, Kazakhstan

5 South Kazakhstan Medical Academy, Shymkent 160000, Kazakhstan

10.22124/cjes.2025.8647

Abstract

A novel gene, AkICE1, and the protein it encodes were molecularly and genetically characterized from Arum korolkowii, a rare endemic species listed in Kazakhstan's Red Book. This plant faces increasing threats from environmental stress, particularly cold stress, exacerbated by climate change. Understanding the genetic mechanisms that underlie cold stress tolerance in such endemic species is critical for conservation and biodiversity preservation. The complete sequence of the AkICE1 gene (2847 bp) was obtained and deposited in GenBank (accession number OR736143). The gene comprises three exons and two introns, encoding a protein of 545 amino acids. Bioinformatic analysis revealed that AkICE1 shares conserved domains with other ICE1 proteins, such as the MYC-like basic helix-loop-helix (bHLH) domain, a serine-rich region (S-rich), and a zipper region (ZIP), all of which are typical of cold stress-related transcription factors. The protein also contains a nuclear localization signal (NLS), suggesting its role in the nucleus as a transcription factor. Additionally, the presence of a sumoylation site, which is crucial for the activation and stability of ICE1 proteins, was identified in AkICE1, indicating that its activity may be regulated by SUMO E3 ligase. The structural analysis predicted 8 alpha helices, 3 beta turns, and 11 coils in the protein, with phosphorylation sites for serine, threonine, and tyrosine residues, further supporting its involvement in cold stress regulation. Phylogenetic analysis revealed that AkICE1 shares the highest sequence similarity with the ICE1 gene of Colocasia esculenta, highlighting its functional conservation within the Aroid family. These findings establish AkICE1 as a novel ICE1-like transcription factor that likely plays a crucial role in cold stress adaptation in A. korolkowii. The study contributes to understanding the molecular mechanisms of cold stress regulation in endemic species, which can aid in developing conservation strategies and breeding programs to improve the resilience of A. korolkowii and other endemic plants to environmental stresses.

Keywords


Badawi, M & Reddy, YM 2008, Structure and functional analysis of wheat ICE (inducer of CBF expression) genes. Plant Cell Physiology, 49: 1237-1249.
Bremer, A & Kent, B 2017, Intrinsically disordered stress protein COR15A resides at the membrane surface during dehydration. Biophysical Journal, 113: 572-579, https://doi.org/10.1016/j.bpj.2017. 06.027.
Chinnusamy, V, Ohta, M, Kanrar, S, Lee, B H, Hong, X, Agarwal, M & Zhu, JK 2003, ICE1: A regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes and Development, 17(8): 1043-1054.
Chinnusamy, V, Zhu, JK, & Sunkar, R 2010, Gene regulation during cold stress acclimation in plants. In JMDLSF Guy (Ed.), Methods in Molecular Biology (Vol. 639, pp. 39-55). Humana Press.
Chinnusamy, V, Zhu, J & Zhu, JK 2007, Cold stress regulation of gene expression in plants. Trends in Plant Science, 12: 444-451.
Ding, Y, Lv, J, Shi, Y, Gao, J, Hua, J, Song, C, Gong, Z & Yang, S 2018, EGR2 phosphatase regulates OST1 kinase activity and freezing tolerance in Arabidopsis. EMBO Journal, 38: e99819.
Guo, J, Ren, Y & Tang, Z 2019, Characterization and expression profiling of the ICE-CBF-COR genes in wheat. PeerJ, 7: e8190.
Hao, X & Wang, L 2018, Stress physiology of tea in the face of climate change. In: Response and adaptation mechanisms of tea plant to low-temperature stress, Springer, pp. 39-61.
Kanaoka, MM, Pillitteri, LJ, Fujii, H, Yoshida, Y, Bogenschutz, NL, Takabayashi, J, Zhu, JK & Torii, KU 2008, SCREAM/ICE1 and SCREAM2 specify three cell-state transitional steps leading to Arabidopsis stomatal differentiation. The Plant Cell, 20: 1775-1785.
Kiegle, E, Moore, CA, Haseloff, J, Tester, MA & Knight, MR 2000, Cell-type-specific calcium responses to drought, salt and cold in the Arabidopsis root. Plant Journal, 23: 267-278.
Kurbidaeva, AS 2015, Study of the role of the ICE2 gene of Arabidopsis thaliana in the control of plant resistance to cold. Doctoral Dissertation, Moscow State University).
Kurbidaeva, A, Novokreshchenova, M & Ezhova, T 2015, ICE genes in Arabidopsis thaliana: Clinal variation in DNA polymorphism and sequence diversification. Biologia Plantarum, 59: 245-252.
Miura, K, Jin, JB, Lee, J, Yoo, CY, Stirm, V, Miura, T, Ashworth, EN, Bressan, RA, Yun, D & Hasegawa, PM 2007, SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis. The Plant Cell, 19: 1403-1414.
Monroy, AF & Dhindsa, RS 1995, Low-temperature signal transduction: Induction of cold acclimation-specific genes of alfalfa by calcium at 25°C. The Plant Cell, 7(3): 321-331.
Nilova, IA 2019, Resistance of wheat plants to high-temperature influences of different intensity: Biophysiochemical and molecular-genetic aspects. Doctoral Dissertation, Petrozavodsk State University, Russia.
Oshunsanya, SO, Nwosu, NJ & Li, Y 2019, Abiotic stress in agricultural crops under climatic conditions. In M. K. Jhariya (Ed.), Sustainable agriculture, forest and environmental management (Vol. 9, pp. 71–100). Springer.
Qing, Y, Wang, S, Yang, Z & Gentine, P 2023, Soil moisture−atmosphere feedbacks have triggered the shifts from drought to pluvial conditions since 1980. Communications Earth & Environment, 4(1): 1-10.
Ritonga, FN & Chen, S 2020, Physiological and molecular mechanism involved in cold stress tolerance in plants. Plants (Basel), 9: 560. https://doi.org/10.3390/plants9050560.
Shu, Y, Li, W, Zhao, J, Zhang, S, Xu, H, Liu, Y, Guo, C 2017, Transcriptome sequencing analysis of alfalfa reveals CBF genes potentially playing important roles in response to freezing stress. Genetics and Molecular Biology, 40: 824-833.
Thomashow, MF 1999, Plant cold acclimation: Freezing tolerance genes and regulatory mechanisms. Annual Review of Plant Physiology and Plant Molecular Biology, 50: 571-599.
Tuteja, N 2007, Abscisic acid and abiotic stress signaling. Plant Signal & Behavior, 2: 135-138.
Yang, C, Yang, H & Xu, Q 2020, Comparative metabolomics analysis of the response to cold stress of resistant and susceptible Tibetan hulless barley (Hordeum distichon). Phytochemistry, 174: 112.
Yeginbay, A, Alpamyssova, G, Yelemanova, Z, Narymbayeva, Z, Daulbay, A, Pernebekova, R, Abduvaliev, BA, Burabaev, A, Burabaev, A & Narimanov, A 2023, Molecular genetic analysis of the ICE1 gene in Arum korolkowii Regel. Caspian Journal of Environmental Sciences, 21: 1143-1149.
Yeginbay, A, Aripova, S, Abubakirova, A, Mutalova, M, Aitkulova, R, Akhmadaliev, BJ, Burabaev, A, Burabaev, A & Narimanov, A 2024, Biology of the medicinal plant Arum korolkowii Regel (Arum). Plant Science Today, 11: 602-605.
Yeginbay, A, Burabaev, A, Burabaev, A, Ibraimova, Z, Yelemanova, Z & Almira, S 2023, Study of the resistance of Arum korolkowii to high-temperature impacts of different intensity at physiological, biochemical, molecular and genetic levels. Indian Journal of Genetics and Plant Breeding, 83: 127-134.
Yeginbay, A, Burabaev, A, Mаmykоvа, R, Burabaev, A, Azhibayeva-Kupenova, D, Zhanar, Y, Anar, E & Zaure, N 2022, Ways and solutions to preserve the gene pool of rare and endangered plants. Caspian Journal of Environmental Sciences, 20: 1135-1142.