Tang Wanhu, Zhang Zuxin, Zou Xiling, Zheng Yonglian
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China.
Sci China C Life Sci. 2005 Aug;48(4):337-45. doi: 10.1360/062004-27.
In this study, SSH (Suppression Subtractive Hybridization) and cDNA microarray were used to identify genes associated with waterlogging response of maize roots. Mo17 and Hz32 are two maize inbred lines with differential tolerance to hypoxia. Seedlings of the inbred lines with two leaves were submerged in hypoxia buffer. SSH libraries were constructed with cDNA samples from roots. Both forward and reverse subtractions were performed for each inbred line, and 105 positive clones induced by hypoxia were selected by differential screening. The treated and control message RNA were hybridized with the cDNA microarray of Mo17, sequentially, 57 of 3-fold differentially expressed clones were obtained. A total of 162 positive clones were all sequenced. Bioinformatics analysis showed these positive clones represent 85 TUGs, including genes involved in several biochemistry pathways, such as glycolysis, protection, signal transduction, cell construction and energy metabolism and 41 EST with unknown function. Comparison between Mo17 and Hz32 indicates that genes related to hypoxia tolerance have different expression patterns in submerged roots. Several positive clones' expression patterns were revealed by Northern or RT-PCR, and a new gene (Sicyp51), which may contribute to hypoxia tolerance, was identified.
在本研究中,利用抑制性消减杂交(SSH)和cDNA微阵列技术来鉴定与玉米根耐涝性相关的基因。Mo17和Hz32是两个对缺氧耐受性不同的玉米自交系。将具有两片叶子的自交系幼苗淹没在缺氧缓冲液中。用根的cDNA样本构建SSH文库。对每个自交系进行正向和反向消减,并通过差异筛选选择105个由缺氧诱导的阳性克隆。将处理后的和对照的信使RNA依次与Mo17的cDNA微阵列杂交,获得57个3倍差异表达的克隆。总共对162个阳性克隆进行了测序。生物信息学分析表明,这些阳性克隆代表85个转录单位组(TUGs),包括参与糖酵解、保护、信号转导、细胞构建和能量代谢等多种生化途径的基因以及41个功能未知的EST。Mo17和Hz32之间的比较表明,与耐缺氧相关的基因在淹水根中有不同的表达模式。通过Northern杂交或RT-PCR揭示了几个阳性克隆的表达模式,并鉴定出一个可能有助于耐缺氧的新基因(Sicyp51)。