| Cloning of MpRZ1 gene in Malus prunifolia and expression analysis of its response to drought stress |
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| DOI：10.7606/j.issn.1000-7601.2024.04.06 |
| Key Words: M. prunifolia RZ gene gene cloning drought stress M. hupehensis gene expression |
| Author Name | Affiliation | | WANG Shuncai | College of Bioengineering and Technology, Tianshui Normal University, Tianshui, Gansu 741000, China
Gansu Key Laboratory for Utilization of Agricultural Solid Waste Resources, Tianshui, Gansu 741000, China | | YANG Zhenling | College of Bioengineering and Technology, Tianshui Normal University, Tianshui, Gansu 741000, China | | TAN Kexin | College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China | | WANG Hongming | College of Bioengineering and Technology, Tianshui Normal University, Tianshui, Gansu 741000, China
Gansu Key Laboratory for Utilization of Agricultural Solid Waste Resources, Tianshui, Gansu 741000, China |
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| Abstract: |
| Based on four RZs transcripts from the transcriptome library of Malus prunifolia under drought stress, a new gene MpRZ1 was cloned by electren extension and RT-PCR methods. The structural and functional characteristics of the deduced amino acid sequence of MpRZ1 gene was analyzed using bioinformatics methods, and a comparative analysis of MpRZ1 protein with predicted apple (Malus domestica) RZ homologous members was conducted. The expression pattern of MpRZ1 gene under drought stress was conducted by qRT-PCR technology. The results indicated that, cDNA sequence of MpRZ1 gene was 930 bp in length contained an intact open reading frame of 843 bp, encoding a polypeptide of 280 amino acid residues with a predicted molecular mass of 30.97 kDa and pI of 9.37. N-terminal signal peptide and transmembrane topology were not found in MpRZ1 protein, which was probably located in the nucleus and an unstable alkaline hydrophilic protein containing 38 potential phosphorylation sites, 13 O-GlcNAc and 5 N-Glyc potential glycosylation sites. The secondary element of MpRZ1 protein was mainly characterized by random curling and α-helice, accounting for up to 87.1%. The predicted MpRZ1 protein contained one conserved RNA-recognition motif (RRM) with five antiparallel β-strands and 2 α-helices arranged in β1α1β2β3α2β4β5 topology at the N-terminus, and a CCHC type zinc finger domain as well as a series of (GGX)n and (DRY/F)n repeated at the C-terminal glycine\|rich region. Sequence alignment and phylogenetic analysis indicated that the predicted MpRZ1 protein shared higher sequence similarity and homology with Arabidopsis and rice RZ proteins, suggesting this MpRZ1 gene belonged to a homologous member of RZ genes family. Nine different MdRZs transcripts found in the apple genome uniformly encoded alkaline hydrophilic proteins, and this MpRZ1 protein had more than 96% homology with two MdRZs proteins encoded by MdP0000088428 and MdP0000272138 transcripts. The expression level of RZ genes in leaves of M. prunifolia and M. hupehensis at 9 days of drought stress and showed significant differences compared to the control (P<0.05), indicating that the MpRZ1 gene was involved in the drought stress response process. |
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