北美鹅掌楸LtCHS基因的克隆及生物信息学与组织表达特征分析

doi:10.11707/j.1001-7488.20150505

Abstract

Abstract:

【Objective】 With obvious inter-specific variation, Liriodendron trees are ideal woody plants for studying gene regulation on flower color. Chalcone synthase genes (CHS) play key roles in the pathway of flavonoid metabolism, and are closely related with plant flower. CHS genes are also ideal for phylogeny study within family due to its large differences between species. Therefore, CHS genes can be used to study the underlying causes for the differences in flower color. In the study, CHS genes were cloned from L. tulipifera, and its bioinformatics and tissue expression were analyzed to provide understanding of the mechanisms of formation and regulation of flower color in Liriodendron. 【Method】 CHS genes (LtCHS) from the flower buds of L. tulipifera were cloned using RACE (rapid amplification cDNA ends) method, and their corresponding genome DNA (gDNA) were also amplified. Based on bioinformatics analysis, semi-quantitative RT-PCR was used to analyze the expression levels of LtCHS genes among different species and different tissues.【Result】The full-length cDNA of LtCHS 1 and LtCHS 2 genes are 875 bp and 1 457 bp, with the open reading frame(ORF)of 702 bp and 1 185 bp respectively. LtCHS 1 gene contains only one exon and encodes 233 amino acids. Its encoding protein has a molecular weight of 71.88 kDa, with theoretical isoelectric point of 5.09. While LtCHS 2 gene consists of two exons and one intron, encoding 394 amino acids. The protein has a molecular weight of 120.0 kDa, with theoretical isoelectric point of 4.97. Analysis of tissue expression of the two genes showed that LtCHS genes had differences not only among tissues of the same species, but also among species. The LtCHS 1 gene was expressed only in L. tulipifera and LtCHS 2 gene was expressed only in L. chinense, while in L. chinense × tulipifera, both genes were expressed.【Conclusion】 The two LtCHS genes obtained in this paper belong to the same family but perform different functions. The expression differences of the LtCHS genes may be associated with color formation of Liriodendron. Considering the characteristics of flower color between species, LtCHS 1 and LtCHS 2 genes may regulate the synthesis of different types of anthocyanins.

Key words: Liriodendron tulipifera, chalcone synthase, gene cloning, semi-quantitative RT-PCR, tissue expression

CLC Number:

S718.46

Luo Qunfeng, Xu Meng, Feng Yuanheng, Xu Jiajuan, Li Huogen. Cloning and Bioinformatics of Chalcone Synthase Gene (CHS) in Liriodendron tulipifera and Characterization of Its Tissue Expression[J]. Scientia Silvae Sinicae, 2015, 51(5): 37-45.

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Cloning and Bioinformatics of Chalcone Synthase Gene (CHS) in Liriodendron tulipifera and Characterization of Its Tissue Expression

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