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利用基于PCR的cDNA扣除杂交技术鉴定小麦杂种与其亲本自交系叶片和根系中的差异表达基因。

Identification of differentially expressed genes in leaf and root between wheat hybrid and its parental inbreds using PCR-based cDNA subtraction.

作者信息

Yao Yingyin, Ni Zhongfu, Zhang Yinhong, Chen Yan, Ding Yuhua, Han Zongfu, Liu Zhiyong, Sun Qixin

机构信息

Department of Plant Genetics and Breeding, Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture/Beijing, China Agricultural University, 100094 Beijing, China.

出版信息

Plant Mol Biol. 2005 Jun;58(3):367-84. doi: 10.1007/s11103-005-5102-x.

Abstract

Heterosis was defined as the advantage of hybrid performance over its parents in terms of growth and productivity. Previous studies showed that differential gene expression between hybrids and their parents is responsible for the heterosis; however, information on systematic identification and characterization of the differentially expressed genes are limited. In this study, an interspecific hybrid between common wheat (Triticum aestivum. L., 2n = 6x = 42, AABBDD) line 3338 and spelt (Triticum spelta L. 2n = 6x = 42, AABBDD) line 2463 was found to be highly heterotic in both aerial growth and root related traits, and was then used for expression assay. A modified suppression subtractive hybridization (SSH) was used to generate four subtracted cDNA libraries, and 748 nonreduandant cDNAs were obtained, among which 465 had high sequence similarity to the GenBank entries and represent diverse of functional categories, such as metabolism, cell growth and maintenance, signal transduction, photosynthesis, response to stress, transcription regulation and others. The expression patterns of 68.2% SSH-derived cDNAs were confirmed by reverse Northern blot, and semi-quantitative RT-PCR exhibited the similar results (72.2%). And it was concluded that the genes differentially expressed between hybrids and their parents involved in diverse physiological process pathway, which might be responsible for the observed heterosis.

摘要

杂种优势被定义为杂种在生长和生产力方面相对于其亲本的杂交表现优势。先前的研究表明,杂种与其亲本之间的基因差异表达是杂种优势的原因;然而,关于差异表达基因的系统鉴定和表征的信息有限。在本研究中,发现普通小麦(Triticum aestivum. L.,2n = 6x = 42,AABBDD)品系3338与斯佩尔特小麦(Triticum spelta L. 2n = 6x = 42,AABBDD)品系2463之间的种间杂种在地上部生长和根系相关性状方面均具有高度杂种优势,随后将其用于表达分析。使用改良的抑制消减杂交(SSH)方法构建了四个消减cDNA文库,共获得748个非冗余cDNA,其中465个与GenBank中的序列具有高度相似性,代表了多种功能类别,如代谢、细胞生长与维持、信号转导、光合作用、应激反应、转录调控等。通过反向Northern杂交验证了68.2%的SSH衍生cDNA的表达模式,半定量RT-PCR也显示出相似的结果(72.2%)。得出的结论是,杂种与其亲本之间差异表达的基因涉及多种生理过程途径,这可能是观察到的杂种优势的原因。

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