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Molecular cloning and characterization of ClZE, a zeaxanthin epoxidase gene in watermelon (Citrullus lanatus)

Authors :
Liu, Yunting
Yao, Daxuan
Hu, Wenjing
Duan, Huijun
Source :
Biotechnology & Biotechnological Equipment; March 2017, Vol. 31 Issue: 2 p259-269, 11p
Publication Year :
2017

Abstract

ABSTRACTIn this study, a watermelon (Citrullus lanatus) zeaxanthin epoxidase gene, ClZE, was isolated by reverse transcription-polymerase chain reaction (PCR) together with RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE). The full cDNA sequence of ClZEis 2535 bp in length containing a 1998 bp open reading frame (ORF) that encodes 665 amino acids. ClZEwas shown to share high homology with the putative ZEgenes in other plant species. Prediction analysis revealed that ClZEbears two large conservative domains, including Pyr_redox and FHA (forkhead-associated domain). Phylogenetic analysis suggested that ClZEshares high similarity (94.8%) with CsZEfrom Cucumis sativus, but is far from the ZEgenes of other species. Prokaryotic expression indicated that ClZE possessed the apparent molecular mass consistent with its calculated molecular mass of 73.1 KDa. Based on quantitative real-time PCR (qRT-PCR), ClZEwas shown to be down-regulated under chilling–low irradiance stress in watermelon leaves. Transgenic Arabidopsislines harbouring ClZEwere generated to test the gene function in mediating plant response to chilling stress. The results indicated that the transgenic lines exhibited decreased non-photochemical quenching (NPQ) and maximum quantum efficiency of photosystem II (PSII) photochemistry (Fv/Fm), increased activities of peroxidase (POD), superoxide dismutase (SOD) and elevated content of malondialdehyde (MDA) relative to the wild type (WT) under chilling–low irradiance stress. In addition, overexpression of ClZEresulted in impaired xanthophyll cycle and aggravated PSII photoinhibition under chilling–low irradiance. All the results together suggested that ClZEplays major roles in regulating the photoinhibition behaviour.

Details

Language :
English
ISSN :
13102818 and 13143530
Volume :
31
Issue :
2
Database :
Supplemental Index
Journal :
Biotechnology & Biotechnological Equipment
Publication Type :
Periodical
Accession number :
ejs41372605
Full Text :
https://doi.org/10.1080/13102818.2016.1275803