热胁迫下丹参转录组cDNA-AFLP分析

cDNA-AFLP analysis on transcriptome of Salvia miltiorrhiza under heat stress

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 research the molecular mechanism of growth and secondary metabolism of Salvia miltiorrhiza during heat stress.Methods The cDNA-AFLP approach was applied to identifying the transcriptome of differential genes expression in leaves.cDNA from leaves was digested by EcoR I and Mse I,and 256 primer combinations were used for selective amplification.Different transcription derived fragments（TDFs） were cloned,sequenced,and analyzed by quantitative RT-PCR.Results TDFs（196） were identified for their differential expression under heat stress,among which the expression of 122 TDFs was enhanced and induced and expression of 74 TDFs was suppressed.Compared the sequences of all TDFs with the NCBI databases,147 TDFs presented homologous,while 49 TDFs with low identity or no match to known genes in NCBI might represent novel genes in S.miltiorrhiza.Conclusion The signal transduction,transcriptional regulation,stress response protein,and metabolism,etc.could be influenced by heat stress,which provides a basis for further study on the regulation of active constituent metabolism of S.miltiorrhiza.