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以生根的“滨梅5号”幼苗为试材,采用cDNA-AFLP差异显示方法研究200mmol/L NaCl处理对基因差异表达的影响,并采用半定量RT-PCR方法验证候选基因的盐胁迫和组织表达模式。结果表明:256对选择性引物组合可扩增出4 263条可分辨的条带,且47条具有明显差异;其中24条经测序后分析表明,14条与已知片段具有较高同源性,主要涉及代谢、转录因子、胁迫相关蛋白等;获取了2条耐逆相关差异基因片段,分别与蝴蝶草天冬酰胺合成酶同源性达92%、与野草莓硫转运蛋白基因同源性达81%;与硫转运蛋白基因同源的差异基因在盐处理3h时表达量最高,随处理时间增加而表达量逐渐下降,在各组织中以根中的表达量最高,推测其主要在根中行使功能。
The effects of 200 mmol / L NaCl treatment on the differential expression of genes were studied by cDNA-AFLP differential display method. The salt stress of candidate gene was verified by semi-quantitative RT-PCR And organizational expression patterns. The results showed that 256 pairs of selectable primer combinations amplified 4 263 distinguishable bands and 47 of them had significant differences. Among them, 24 were sequenced and analyzed, and 14 showed high homology with known fragments, mainly Related genes, such as metabolism, transcription factors and stress-related proteins; obtained two resistance-related differential gene fragments, respectively, homology with butterflies asparagine synthase 92%, wild strawberry sulfur transporter gene homology of 81% ; Genes with homology to the sulfur transporter gene expressed the highest at 3h after salt treatment and gradually decreased with the increase of treatment time, and the highest expression level in roots was observed in each tissue, suggesting that it mainly functions in roots .