Molecular cloning and characterization of a NAC-like gene in "navel" orange fruit response to postharvest stresses
Molecular cloning and characterization of a NAC-like gene in "navel" orange fruit response to postharvest stresses
复制标题
脐橙果实对采后应激反应中的 NAC 类基因的分子克隆和特征描述
DOI：
10.1007/s11105-007-0016-1
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发表时间：
2007-12-01
影响因子：
2.1
通讯作者：
Li, Zheng-Guo
中科院分区：
文献类型：
--
作者：
Fan, Jing;Gao, Xue;Li, Zheng-Guo
构建了柑橘果皮凹陷差异表达基因的消减文库。根据与NAC基因家族同源的一个基因片段的序列，采用快速扩增的方法，从脐橙中克隆了1,203个核苷酸的全长cDNA。它被命名为CsNAC，编码305个氨基酸的蛋白质。计算得到CsNAC蛋白的相对分子质量为35.2 kDa，理论等电点为6.72。序列比较表明，CsNAC蛋白在N端有一个明显的保守区，这被认为是NAC蛋白家族的特征。CsNAC蛋白与拟南芥ATAF1同源。系统发育分析证实，CsNAC属于ATAF亚家族，在应激反应中发挥重要作用。RNA凝胶印迹分析表明，CsNAC基因在创伤和无氧等胁迫下迅速而强烈地诱导表达。低温(4℃)和乙烯暴露也增加了CsNAC基因的表达水平。然而，它的表达被高温(40℃)抑制，但不受低氧(3%)影响。我们的研究结果为进一步研究类NAC基因在逆境诱导柑橘果皮凹陷中的作用奠定了基础。
A cDNA subtraction library had been constructed to identify differentially expressed genes in peel pitting of citrus fruit. Based on the sequence of a cDNA fragment homologous to NAC gene family, the full-length cDNA of 1,203 nucleotides was cloned from "navel" orange by rapid amplification of cDNA ends. It was designated as CsNAC, encoding a protein of 305 amino acids. The calculated molecular weight of the CsNAC protein was 35.2 kDa, and theoretical isoelectric point was 6.72. Sequence comparison showed that the CsNAC protein had a strikingly conserved region at the N terminus, which is considered as the characteristic of the NAC protein family. CsNAC protein was orthologous to Arabidopsis thaliana ATAF1. Phylogenetic analysis confirmed CsNAC belonged to the ATAF subfamily, which plays an important role in response to stress stimuli. RNA gel blot analysis showed that the expression of CsNAC gene was rapidly and strongly induced by stresses such as wounding and no oxygen. Low temperature (4 degrees C) and exposure to ethylene also increased the expression level of CsNAC gene. However, its expression was suppressed by high temperature (40 degrees C) but not affected by low oxygen (3%). Our results may provide the basis for future research of NAC-like gene's role in stress-induced citrus peel pitting.