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Archive > Volume 15 Issue 4 > 2023,15(4):574-587. DOI:https://doi.org/10.1016/j.chmed.2022.08.008 Prev Next

防風(fēng)轉(zhuǎn)錄組分析及抽薹開花基因的挖掘

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Transcriptome analysis of Saposhnikovia divaricata and mining of bolting and flowering genes

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    摘要:
    目的：早期抽薹嚴(yán)重影響了防風(fēng)藥用價(jià)值和資源的可持續(xù)開發(fā)。防風(fēng)抽薹和開花的分子機(jī)制尚不清楚，值得進(jìn)一步研究。在我們的研究中，我們探索了與防風(fēng)抽薹和開花相關(guān)基因的轉(zhuǎn)錄組學(xué)。 方法：采用高通量測序方法，從現(xiàn)蕾開花期的防風(fēng)未抽薹和抽薹葉片中構(gòu)建轉(zhuǎn)錄組文庫，并對(duì)其進(jìn)行測序、組裝和注釋。專注于與植物抽薹和開花有關(guān)的途徑，并探索基因。通過實(shí)時(shí)熒光定量PCR (qRT-PCR)檢測7個(gè)候選基因的表達(dá)。 結(jié)果：轉(zhuǎn)錄組學(xué)結(jié)果顯示，獲得了249,889,422個(gè)高質(zhì)量的clean reads。共組裝67866個(gè)unigenes，平均長度948.1 bp。Trinity de Novo組裝共產(chǎn)生67,866個(gè)unigenes，平均長度為948.1 bp。在SdM組993個(gè)差異表達(dá)基因中，有484個(gè)基因表達(dá)顯著上調(diào)，509個(gè)基因表達(dá)顯著下調(diào)。共有79個(gè)GO條目在差異表達(dá)基因中顯著富集。KEGG結(jié)果顯示，11,154個(gè)unigenes富集在89條通路中。同時(shí)，挖掘了21個(gè)與防風(fēng)抽薹和開花相關(guān)的候選基因。qRT-PCR結(jié)果顯示，HDA9、PHYB、AP2、TIR1、Hsp90、CaM、IAA7的表達(dá)趨勢與轉(zhuǎn)錄組測序結(jié)果一致。此外，RNA-seq還鑒定出10,740個(gè)轉(zhuǎn)錄因子，并根據(jù)其保守結(jié)構(gòu)域?qū)⑵浞譃?8個(gè)家族。進(jìn)一步研究表明，調(diào)控防風(fēng)開花的轉(zhuǎn)錄因子主要分布在NAC、MYB_related、HB-other、ARF和AP2家族。 結(jié)論：本研究結(jié)果表明，植物激素信號(hào)轉(zhuǎn)導(dǎo)通路是控制抽薹和開花的決定性因素之一。其中，生長素相關(guān)基因IAA和TIR1是影響防風(fēng)抽薹和開花過程的關(guān)鍵基因。

    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.

    關(guān)鍵詞:
    抽薹與開花;比較分析;開花基因;防風(fēng);轉(zhuǎn)錄組學(xué)

    Keywords:


    Project Supported:
    This work was the supported by China Agriculture Research System of MOF and MARA (No. CARS-21), which are gratefully acknowledged.

    Clc Number:

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