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Identification of EgrZFP-EARs of the C2H2 Zinc Finger Protein Gene Family in Eucalyptus grandis and Analysis of Their Expression Pattern Under Abiotic Stresses |
YU Jian-Feng, HONG Jia-Du, ZHAO Shuang, LI Xiao-Mei, WEN Dan-Dan, CHENG Long-Jun* |
College of Forestry and Biotechnology/State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China |
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Abstract Eucalyptus spp. are widely planted in south of China as a fast-growing timber species. However, the resistance to abiotic stresses such as low temperature, drought, and salinity of them is very poor. Zinc finger protein (ZPF) transcription factor members containing the ethylene-responsive element binding factor-associated amphiphilic repression (EAR) motif have been shown to be extensively involved in plant responses to abiotic stresses such as cold, high temperature, drought and salinity. Based on the genome-wide identification of Eucalyptus grandis C2H2-type ZFP, the EgrZFP-EAR genes with EAR motif was screened. The evolutionary relationships of EgrZFP-EAR, the types, numbers and distributions of EAR motifs in their proteins, chromosome localization and gene replication of them, as well as gene and protein sequence structure and the cis-elements on their promoters were analyzed. In addition, transcriptome data and qRT-PCR were used to analyze the expression patterns of EgrZFP-EAR in different tissues and 4 ℃, high temperature (42 ℃), drought, NaCl (300 mmol/L), ABA (100 μmol/L) and MeJA (100 μmol/L) treatments of E. grandis seedlings. The results showed that there were 118 C2H2-type ZFP in E. grandis, 57 of which were EgrZFP-EAR members, and 54 of these members encoded ZFP proteins containing Q-type sequence (QALGGH). They contained 3 types of EAR motifs: LxLxL, DLNx (x) p, and their overlap. The LxLxL type had the largest number of them, with a total of 98. Evolutionarily, genome-wide replication, tandem duplication and segment duplication all played roles in the expansion of these genes. Stress responsive elements such as DRE, MBS, LTR and STRE, and hormone responsive elements including ABRE and CGTCA-motif were distributed on the EgrZFP-EAR promoters. Tissue expression patterns were divided into 4 types: high expression in shoot tips and young leaves; high expression in phloem; high expression in mature xylem; and high expression in phloem, immature and mature xylem. The expression patterns of EgrZFP-EAR gene in leaves of E. grandis seedlings treated with 4 ℃ for different time (2, 6, 12, 24, 48 h) were classified into 3 types. Genes in the first type were induced persistently; the expression of genes in the second type ascended firstly and then descended; and in the third type, genes' expression was mainly inhibited. The selected 12 genes from different EgrZFP-EAR subgroup were also responsive differently to high temperature, drought, NaCl, ABA and MeJA treatments, implicating the complex function of EgrZFP-EAR gene under abiotic stresses. The results provide an foundation for further revealing the molecular mechanism of stress resistance, the exploitation of stress-resistant gene resources and molecular-assisted stress resistance breeding of Eucalyptus.
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Received: 20 August 2023
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Corresponding Authors:
* ljcheng@zju.edu.cn
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