团花树α扩展蛋白基因的克隆及表达分析

摘要/Abstract

摘要：

采用扩增保守区域、基因组步移及3'RACE技术,在团花树中克隆到15个α-expansin基因的cDNA序列,命名为AcEXPA2-16(GenBank注册号JF922686-JF922700),其相对应的基因组序列GenBank注册号为JF922701-JF922715。包括已知的AcEXPA1,这16个EXPA蛋白的大小及序列高度保守,包括N端的信号肽、3个外显子和2个内含子。通过氨基酸序列比对和系统进化树分析,AcEXPA1-16分为4个亚族,分别为A、B、C和D亚族。定量PCR分析显示AcEXPA1-16基因在同一组织中的表达存在冗余现象,而单个基因在不同的组织中又存在特异表达,尤其AcEXPA8在形成层中的表达水平极高。这表明AcEXPA8在木质部发生的过程中可能起到重要的作用。研究结果为在团花树中研究α-expansin基因与木质部的生长和材质的关系提供信息,并最终为林木分子育种提供潜在的候选基因。

关键词: 团花树, &alpha, 扩展蛋白, 基因克隆, 定量PCR

Abstract:

Fifteen cDNAs, designated as AcEXPA 2 to AcEXPA16 (GenBank accession No. JF922686-JF922700) with the corresponding genomic DNA sequences (GenBank accession No. JF922701-JF922715), were isolated by amplifying conserved domain binding with genomic walking and 3'RACE techniques from four differential tissues in Anthocephalus chinensis. The size and sequence of these α-expansin homologues including known AcEXPA1 were highly conserved, and had the same sequence structures as an N-terminal signal peptide, three exons and two introns. Their amino acid alignment and phylogenetic analysis showed that A. chinensis α-expansin genes were divided into four subgroups: A, B, C and D. A real-time quantitative PCR analysis showed that the transcript level of AcEXPA 1-16 was redundant in the same tissue and tissue-specific with each gene in different tissue, especially, the transcript level of AcEXPA 8 extremely high in the cambium region. These results suggest that AcEXPA 8 may play an important role in xylogenesis. This study on α-expansin genes from A. chinensis will lay the foundation for studying the relationship between α-expansin genes and growth rate and wood quality of the xylem in this fast-growing tree, and finally provide potential candidate genes for tree molecular breeding.

Key words: Anthocephalus chinensis, α-expansin, gene cloning, RT-qPCR

中图分类号:

S718.46

欧阳昆唏, 李俊成, 黄浩, 刘明骞, 陈晓阳. 团花树α扩展蛋白基因的克隆及表达分析[J]. 林业科学, 2013, 49(9): 62-71.

Ouyang Kunxi, Li Juncheng, Huang Hao, Liu Mingqian, Chen Xiaoyang. Molecular Cloning and Expression Analysis of α-Expansin Genes in Anthocephalus chinensis[J]. Scientia Silvae Sinicae, 2013, 49(9): 62-71.

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