Transcriptome analysis of Saposhnikovia divaricata and mining of bolting and flowering genes
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摘要:
目的：早期抽薹严重影响了防风药用价值和资源的可持续开发。防风抽薹和开花的分子机制尚不清楚，值得进一步研究。在我们的研究中，我们探索了与防风抽薹和开花相关基因的转录组学。 方法：采用高通量测序方法，从现蕾开花期的防风未抽薹和抽薹叶片中构建转录组文库，并对其进行测序、组装和注释。专注于与植物抽薹和开花有关的途径，并探索基因。通过实时荧光定量PCR (qRT-PCR)检测7个候选基因的表达。 结果：转录组学结果显示，获得了249,889,422个高质量的clean reads。共组装67866个unigenes，平均长度948.1 bp。Trinity de Novo组装共产生67,866个unigenes，平均长度为948.1 bp。在SdM组993个差异表达基因中，有484个基因表达显著上调，509个基因表达显著下调。共有79个GO条目在差异表达基因中显著富集。KEGG结果显示，11,154个unigenes富集在89条通路中。同时，挖掘了21个与防风抽薹和开花相关的候选基因。qRT-PCR结果显示，HDA9、PHYB、AP2、TIR1、Hsp90、CaM、IAA7的表达趋势与转录组测序结果一致。此外，RNA-seq还鉴定出10,740个转录因子，并根据其保守结构域将其分为58个家族。进一步研究表明，调控防风开花的转录因子主要分布在NAC、MYB_related、HB-other、ARF和AP2家族。 结论：本研究结果表明，植物激素信号转导通路是控制抽薹和开花的决定性因素之一。其中，生长素相关基因IAA和TIR1是影响防风抽薹和开花过程的关键基因。
Abstract:
Objective: Early bolting of Saposhnikovia divaricata has seriously hindered its medicinal value and sustainable development of resources. The molecular mechanism of bolting and flowering of S. divaricata is still unclear and worth of research. In our study, we explored the transcriptome of the genes related to the bolting and flowering of S. divaricata. Methods: The transcriptome library was constructed, sequenced, assembled and annotated from the bolting and unbolting leaves of S. divaricata by high-throughput sequencing at the bud and flowering stage. Focus on the pathways related to bolting and flowering in plants, and explore genes. The expression of seven candidate genes was verified by real-time fluorescence quantitative PCR (qRT-PCR). Results: Transcriptome results showed that 249 889 422 high-quality clean reads were obtained. A total of 67 866 unigenes were assembled with an average length of 948.1 bp. Trinity de Novo assembly produced 67 866 unigenes with an average length of 948.1 bp. Among 993 differentially expressed genes, 484 genes were significantly up-regulated and 509 genes were down-regulated in the SdM group. A total of 79 GO terms were significantly enriched for differentially expressed genes. KEGG results showed that 11 154 unigenes were enriched in 89 pathways. And 21 candidate genes related to bolting and flowering of S. divaricata were excavated. The qRT-PCR results showed that expression trends of HDA9, PHYB, AP2, TIR1, Hsp90, CaM, and IAA7 were consistent with transcriptomic sequencing results. In addition, RNA-seq had identified 10 740 transcription factors and classified them into 58 families by their conserved domains. Further studies showed that the transcription factors regulating the flowering of S. divaricata were mainly distributed in the NAC, MYB_related, HB-other, ARF, and AP2 families. Conclusion: Based on the results of this study, it was found that the plant hormone signal transduction pathway was one of the decisive factors to control bolting and flowering. Among them, auxin related genes IAA and TIR1 are the key genes in the bolting and flowering process of S. divaricata.
关键词:
抽薹与开花;比较分析;开花基因;防风;转录组学
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This work was the supported by China Agriculture Research System of MOF and MARA (No. CARS-21), which are gratefully acknowledged.
Min Zhang a, b,, Wenle Wang a, c,, Qian Liu b, Erhuan Zang b, Lijun Wu d, Guofa Hu d, Minhui Li a, b,c,*. Transcriptome analysis of Saposhnikovia divaricata and mining of bolting and flowering genes[J]. Chinese Herbal Medicines (CHM),2023,15(4):574-587
