Identification of genes preferentially expressed in cotton fibers: A possible role of calcium signaling in cotton fiber elongation
Identification of genes preferentially expressed in cotton fibers: A possible role of calcium signaling in cotton fiber elongation
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DOI：
10.1016/j.plantsci.2007.04.008
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发表时间：
2007-07-01
期刊：
影响因子：
5.2
通讯作者：
Xia, Gui-Xian
中科院分区：
文献类型：
--
作者：
Gao, Peng;Zhao, Pi-Ming;Xia, Gui-Xian
棉纤维是一种极细长的单细胞，是研究植物细胞伸长机制的理想系统。本研究利用棉花叶片RNA和纤维RNA进行抑制性消减杂交（SSH），筛选纤维细胞特异表达或优先表达的基因。通过反向北方分析筛选得到的SSH文库，鉴定出总共180个差异表达的cDNA片段。序列测定和数据库分析显示，64个非冗余cDNA克隆，其中25个编码未知功能的蛋白质。在这些cDNA中，发现四个编码与钙信号传导组分显示出高度同源性的蛋白质，包括钙调蛋白（CaM）、谷氨酸脱羧酶（GAD）和钙调神经磷酸酶B样（CBL）蛋白相互作用蛋白激酶（CIPKs）。RT-PCR分析表明，这些基因的转录本主要积累在伸长纤维细胞。此外，与野生型对照相比，这些基因的表达水平在li（无配体）突变体纤维中显著降低。两种CIPK中的一种，命名为GhCIPK 1，在本研究中进一步表征。结构分析表明，GhCIPK 1基因含有CIPK蛋白的特征结构域，在纤维发育的延伸期高表达，体外分析表明该基因产物是一种功能性蛋白激酶。我们的研究结果表明，钙介导的信号转导可能在棉纤维伸长中起重要作用。(c)2007爱思唯尔爱尔兰有限公司保留所有权利。
Cotton fiber is an extremely elongated single cell that i considered as an ideal system for studying the mechanisms controlling plant cell elongation. In this study, suppression subtractive hybridization (SSH) between RNA from leaves and fibers of cotton plant was conducted to identify genes that are specifically or preferentially expressed in fiber cells. Screening the resulting SSH library by reverse Northern analysis identified a total of 180 differentially expressed cDNA fragments. Sequencing determination and database analysis revealed 64 non-redundant cDNA clones, of which, 25 code for unknown-function proteins. Among these cDNAs, four were found to encode for proteins that showed high homology to calcium signaling components including calmodulin (CaM), glutamate decarboxylase (GAD) and calcineurin B-like (CBL) protein-interacting protein kinases (CIPKs). RT-PCR analysis indicated that the transcripts of these genes were accumulated predominantly in elongating fiber cells. Moreover, the expression level of these genes was significantly reduced in the li (ligon-lintless) mutant fibers as compared to the wildtype control. One of the two CIPKs, designated as GhCIPK1, was further characterized in this study. Structural analysis showed that GhCIPK1 contained the characteristic domains of CIPK proteins and was highly expressed in the elongating phase in developing fiber, and in vitro assay demonstrated that the gene product was a functional protein kinase. Our results suggest that calcium-mediated signal transduction may play an important role in cotton fiber elongation. (c) 2007 Elsevier Ireland Ltd. All rights reserved.