毛竹4个B-box锌指蛋白序列和基因表达特征及PeBZF4的功能

doi:10.11707/j.1001-7488.20150305

Abstract

Abstract:

【Objective】 Zinc finger proteins containing B-box (BZF) play important roles in regulating growth and development, and response to stresses in plants. Light stress has a significant impact on the growth and development of bamboo. Studies on protein sequence of BZF and expression profiles of BZF genes in Phyllostachys edulis and fluorescence parameters of transgenic plants with over expressed PeBZF 4 were conducted in order to reveal the response and regulatory mechanisms of BZF genes in bamboo under high intensity of light.【Method】Homologous gene alignment method was used to obtain bamboo BZF sequences from BambooGDB directly. The analysis and comparison of gene and protein, the prediction of acting elements, protein functional domains, hydrophilicity and hydrophobicity of proteins were performed using special software and online public platform. Real-time quantitative PCR was used for the analysis of gene expression in different tissues, and the changes of gene expression in leaf after the treatments of high intensity of light (1 200 μmol·m^-2s^-1) and darkness. The over expression vector of PeBZF 4 was constructed and transferred into Arabidopsis thaliana mediated by Agrobacterium tumefanciens. The phenotype of transgenic plants was observed directly and the fluorescence parameters were measured using IMAGING-PAM.【Result】Four homologue BZF genes were obtained from BambooGDB, and designed as PeBZF 1, PeBZF2, PeBZF3 and PeBZF4 respectively. Cis-acting element analysis showed that four PeBZFs all had light-responsive cis-acting regulatory elements and light responsive elements, but the light-responsive elements were different, including MNF1 and Sp1 in PeBZF 1, I-box, Sp1 and boxⅡin PeBZF 2, GT1-motif, MNF1 and Sp1 in PeBZF 3 and PeBZF 4. The analysis of protein structure showed that the proteins encoded by these genes all contained two B-box domains and CHC3H2 binding motif, indicating that they were B-box zinc finger proteins. Tissue specific expression indicated that the levels of these PeBZFs were obviously different in leaf, sheath, stem and root, among which the highest level of PeBZF 1 and PeBZF3 was found in leaf, and that of PeBZF 2 and PeBZF 4 was in sheath. All the expression of these PeBZFs were inhibited by high intensity of light (1 200 μmol·m^-2s^-1). The expression of PeBZF 2 and PeBZF3 showed a gradual downward trend with the extension of irradiation time, while that of PeBZF 1 and PeBZF4 raised slightly after 1 h treatment compared with 0.5 h treatment, followed by a rapid decline. The expression of PeBZF 1, PeBZF2 and PeBZF3 were significantly inhibited by darkness, while that of PeBZF 4 was significantly upregulated about 3.2 times of that in control. The actual photosynthetic efficiency (Y(Ⅱ)) and non-photochemical quenching (NPQ) of transgenic plants with over expressed PeBZF 4 were higher than those of wild type.【Conclusion】 There were four different B-box-type zinc finger protein genes expressed constitutively in Ph. edulis, the expression changes of 4 genes under high intensity of light and dark treatments suggested that they were involved in regulation of light response to stress. The values of Y (Ⅱ) and NPQ increased in transgenic plants with over expressed PeBZF 4, indicating that PeBZF 4 could improve the thermal dissipation capability and photochemical efficiency of transgenic plants. This result indicated that BZF might be associated with bamboo resistance to light stress.

Key words: Phyllostachys edulis, B-box zinc finger protein gene, light stress, gene expression

CLC Number:

S718.46

Lou Yongfeng, Yang Li, Peng Zhenhua, Zhao Hansheng, Gao Zhimin. Protein Sequences and Expression Profiles of 4 B-box Zinc Finger Genes and Functions of PeBZF 4 from Phyllostachys edulis[J]. Scientia Silvae Sinicae, 2015, 51(3): 34-40.

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Protein Sequences and Expression Profiles of 4 B-box Zinc Finger Genes and Functions of PeBZF 4 from Phyllostachys edulis

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