Cloning and Functional Analysis of Low Temperature Response Gene,GhZAT10, in Upland Cotton at Seedling Stage |
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Authors: | Yang Yongfei Ge Changwei Shen Qian Zhang Siping Liu Shaodong Ma Huijuan Chen Jing Liu Ruihua Li Shicong Zhao Xinhua Li Zhikun Pang Chaoyou |
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Affiliation: | 1.Hebei Agricultural University/Hebei Base of State Key Laboratory of Cotton Biology, Baoding, Hebei 071001, China; 2.Institute of Cotton Research of the Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology, Anyang, Henan 455000, China |
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Abstract: | [Objective] The aim of this study is to identify chilling tolerant genes which can provide a basis for breeding upland cotton cultivars with tolerance to chilling. [Method] The GhZAT10 (Zinc finger of Arabidopsis thaliana 10) gene was cloned, and its expression in roots, stems, and leaves was analyzed with cold treatment by quantitative real-time polymerase chain reaction (qRT-PCR). Then we analyzed the structural characteristics of GhZAT10, protein properties and constructed the phylogenetic tree by bioinformatics methods. Next, we constructed subcellular localization vector 35S::GhZAT10-GFP by Gateway technology. Finally, we studied the effect of GhZAT10 in chilling response by virus induced gene silencing of cotton. [Result] The Open reading frame(ORF) length of GhZAT10 gene is 813 bp (base pair), encoding 270 amino acid residues. The expression of GhZAT10 in roots was higher than that in stems and leaves, and up-regulated after chilling stress in these tissues. GhZAT10 does not possess signal peptides or transmembrane helices; its activity is closely related to the phosphorylation regulation. GhZAT10 has homology with the ZAT10 of Theobroma cacao, Arabidopsis thaliana and Citrus sinensis. GhZAT10 protein is located in the nucleus. The GhZAT10-silenced cotton plants were more sensitive to low temperature than wild type. [Conclusion] GhZAT10 belongs to C2H2 zinc finger protein, and plays a positive role to response the chilling stress in upland cotton. |
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