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目的探索热胁迫影响丹参生长和次生代谢的分子机制。方法采用cDNA-AFLP技术对丹参热胁迫下叶片基因表达进行转录组分析。采用EcoR I和Mse I对叶片cDNA进行双酶切,256对引物组合进行选择性扩增,对差异片段进行测序、功能分析和定量RT-PCR。结果共分离出196个差异表达转录衍生片段(transcription derived fragments,TDFs),其中增强与诱导表达的TDFs 122条,抑制表达的74条;从NCBI中鉴定出147个同源TDFs,另有49个TDFs为新发现序列。结论热胁迫影响转录组的信号转导、转录调控、逆境响应蛋白、物质代谢等多个方面,为进一步研究丹参有效成分的代谢调控奠定了基础。
Objective To explore the molecular mechanism of heat stress on the growth and secondary metabolism of Salvia miltiorrhiza. Methods cDNA-AFLP was used to analyze the gene expression of leaves under heat stress of Salvia miltiorrhiza. Double digestion of leaf cDNA with EcoR I and Mse I, selective amplification of 256 primer combinations, sequencing of differential fragments, functional analysis and quantitative RT-PCR. Results A total of 196 differentially expressed transcription derived fragments (TDFs) were isolated, of which 122 were enhanced and induced by TDFs and 74 were inhibited. 147 homologous TDFs were identified in NCBI and 49 TDFs are newly discovered sequences. Conclusion Heat stress can affect signal transduction, transcriptional regulation, stress response protein and substance metabolism in transcriptome, which lays the foundation for the further study on the metabolic regulation of the active ingredients of Radix Salviae Miltiorrhizae.