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【目的】研究甘蔗应答水分胁迫差异基因表达,进一步探究甘蔗应答水分胁迫的分子机制,为开展甘蔗抗旱综合性遗传改良研究提供科学依据。【方法】选用GT11为材料,分别构建对照、干旱、复水3个处理总RNA混合池,使用cDNA-SCoT差异显示构建GT11伸长期应答水分胁迫的基因表达谱,筛选、分离TDFs进行测序,在NCBI数据库进行BLAST同源性检索,根据同源基因推测基因功能。【结果】通过42条SCoT引物筛选得到180个TDFs,有100个TDFs与NCBI数据库中已录入的基因具有较高的相似性,根据同源基因功能可分为14类:新陈代谢相关基因(11个)、能量代谢相关基因(12个)、运输途径相关基因(5个)、通信及信号转导相关基因(6个)、细胞循环及DNA加工相关基因(3个)、转录调控因子相关基因(3个)、蛋白质合成相关基因(17个)、蛋白质加工相关基因(1个)、结合功能蛋白相关基因(7个)、环境互作相关基因(3个)、转座子、毒性及质粒蛋白相关基因(5个)、细胞成分生物合成相关基因(1个)、防御相关基因(4个)和未知功能蛋白基因(22个)。【结论】甘蔗应答水分胁迫调节途径具有多样性、复杂性、协调性和网络性;应用cDNA-SCoT差异显示筛选获得水分胁迫相关TDFs,可为进一步研究甘蔗应答水分调控途径的分子机理提供重要信息。
【Objective】 The objective of this study was to investigate the differential gene expression of sugarcane in response to water stress and to further explore the molecular mechanism of sugarcane response to water stress and to provide a scientific basis for the study of comprehensive genetic improvement of sugarcane drought resistance. 【Method】 GT11 was used as material to construct three pools of total RNA mixed with control, drought and rehydration respectively. CDNA-SCoT was used to construct the gene expression profile of response to water stress in GT11 elongation stage. The TDFs were screened and sequenced. NCBI database for BLAST homology search, infer the function of genes according to homologous genes. 【Result】 Totally 180 TDFs were screened by 42 SCoT primers, and 100 TDFs were highly similar to those already recorded in the NCBI database. According to the function of homologous genes, they could be divided into 14 categories: 11 genes ), 12 genes related to energy metabolism, 5 genes related to transport pathway, 6 genes related to communication and signal transduction, 3 genes related to cell cycle and DNA processing, 3 genes related to transcriptional regulatory factors 3), 17 genes involved in protein synthesis, 1 gene related to protein processing, 7 genes related to functional protein (3), 3 genes related to environmental interaction, transposons, virulence and plasmid proteins Related genes (5), cell biosynthesis related genes (1), defense-related genes (4) and unknown function protein genes (22). 【Conclusion】 The diversity, complexity, coordination and network of sugarcane response water stress regulation pathways are diverse. The application of cDNA-SCoT differential display to screen for water stress-related TDFs may provide important information for further studying the molecular mechanism of sugarcane response to water regulation .