低磷胁迫柱花草差异表达基因分析研究

Analysis of Differentially Expressed Genes in Low Phosphorus Stress of Stylosanthes Guianensis

研究柱花草在低磷条件下生长的分子机理和反应机制。采用cDNA-SRAP分子标记技术分离柱花草茎和叶在低磷胁迫下相关基因的差异表达片段,并对其进行生物信息学分析。结果表明,8对引物组合共扩增出差异片段195条,其中抑制型表达片段56条,诱导型表达片段92条,上调表达型差异片段36条,下调表达型片段11条,大小均在50~1000 bp之间。二次扩增后将特异性条带回收后进行测序,通过BLAST比对,共比对出14条差异片段的同源序列,其中8个差异表达的核苷酸序列与已知功能基因(JCVI-FLLj-1K4未知mRNA、NBS-LRR型抗病蛋白、ATP合成酶、醛固酮还原酶、叶绿体RF2)有较高同源性,5个序列与假定基因或预测基因(UPF0051蛋白、蛋白酶启动子、SCEI结合酶、吲哚-3-丙酮酸盐单氧酶)同源性较高。本研究为筛选柱花草响应低磷胁迫的差异基因提供参考。

The cDNA-SRAP molecular marker technique was used to isolate the differentially expressed genes of the stems and leaves of Stylosanthes under low phosphorus stress, and bioinformatics analysis was carried out to study the molecular mechanism and reaction mechanism of Stylosanthes growing under low phosphorus condition. A total of 195 differential fragments were amplified from 8 pairs of primer combinations. Among them, there were 56 inhibitory expression fragments, 92 inducible expression fragments, 36 upregulated expression fragments and 11 down-regulated expression fragments, and the sizes ranged from 50 to 1000 bp. After the second amplification, the specific bands were recovered and sequenced. Through blast alignment, the homologous sequences of 14 different fragments were compared, and the 8 differentially expressed nucleotide sequences had high homology with known functional genes(JCVI-FLLj-1 K4 unknown mRNA, NBS-LRR type disease resistance protein, ATP synthase, aldosterone reductase, chloroplast RF2), five sequences were highly homologous to the putative or predicted genes(UPF0051 protein, protease promoter, SCEI binding enzyme, indole-3-pyruvate monooxygenase). This study has laid a foundation for screening differential genes in response to low phosphorus stress.