[關(guān)鍵詞]
[摘要]
目的 對(duì)甲基磺酸乙酯(ethyl methanesulfonate,EMS)誘變下蒲公英Taraxacum mongolicum胚性細(xì)胞進(jìn)行轉(zhuǎn)錄組測(cè)序,探究EMS誘變對(duì)蒲公英基因表達(dá)的影響。方法 利用Illumina HiSeq 2500高通量測(cè)序技術(shù)對(duì)對(duì)照(CK)、0.15% EMS處理8 h后的蒲公英胚性細(xì)胞進(jìn)行測(cè)序并建立cDNA數(shù)據(jù)庫(kù),經(jīng)拼接后得到Unigene,并進(jìn)一步開(kāi)展生物信息學(xué)分析。結(jié)果 共獲得Unigene 45 551條,分別有32 954、12 923、21 752、28 115、9 764條被非冗余數(shù)據(jù)庫(kù)(non-redundant protein sequence database,NR)、基因本體(gene ontology,GO)、Swiss-Prot、eggNOG、京都基因與基因組數(shù)據(jù)庫(kù)(Kyoto encyclopedia of genes and genomes,KEGG)等數(shù)據(jù)庫(kù)注釋?zhuān)珿O富集分析發(fā)現(xiàn),蒲公英胚性細(xì)胞的DEGs在GO功能注釋中主要集中于細(xì)胞過(guò)程、代謝過(guò)程、結(jié)合蛋白、氧化還原、催化活性等功能。KEGG途徑富集結(jié)果表明,EMS誘變下DEGs顯著富集在植物激素信號(hào)轉(zhuǎn)導(dǎo)、光合作用、光合生物的固碳作用及光合作用-天線(xiàn)蛋白、胡蘿卜素合成、不飽和脂肪酸代謝以及淀粉和糖代謝途徑。EMS誘變能引起蒲公英胚性細(xì)胞植物激素信號(hào)轉(zhuǎn)導(dǎo)途徑關(guān)鍵酶基因的表達(dá)上調(diào),響應(yīng)脅迫。結(jié)論 通過(guò)轉(zhuǎn)錄組測(cè)序,初步表明EMS誘變下蒲公英胚性細(xì)胞差異基因的表達(dá),并調(diào)控蒲公英胚性細(xì)胞中植物激素信號(hào)轉(zhuǎn)導(dǎo)途徑的相關(guān)酶基因,以期為從分子基因方面培育優(yōu)質(zhì)蒲公英提供參考和依據(jù)。
[Key word]
[Abstract]
Objective Transcriptome sequencing was performed on Taraxacum mongolicum embryogenic cells induced by ethyl methanesulfonate to explore the effect of ethyl methanesulfonate mutagenesis on gene expression in T. mongolicum. Methods Illumina HiSeq 2500 high-throughput sequencing technology was used to sequence embryogenic cells of T. mongolicum in 0% EMS (control) and 0.15% EMS treatments for 8 h and establish cDNA database. Unigene was obtained by splicing, and further biochemistry analysis was carried out. Results A total of 32 954, 12 923, 21 752, 28 115 and 9 764 Unigene were annotated by non-redundant protein sequence database (NR), gene ontology (GO), Swiss-Prot, eggNOG and Kyoto Encyclopedia of genes and genomes (KEGG) databases, respectively. A total of 45 551 DEGs were screened. GO enrichment analysis revealed that the DEGs of dandelion embryonic cells were mainly concentrated in cellular processes, metabolic processes, binding proteins, redox and catalytic activity in GO functional annotation. The KEGG pathway enrichment results showed that DEGs were significantly enriched in plant hormone signal transduction, photosynthesis, carbon sequestration in photosynthetic organisms, photosynthesis antenna protein synthesis, carotenoid synthesis, unsaturated fatty acid metabolism, and starch and sugar metabolism pathways under EMS mutagenesis. EMS mutagenesis can induce upregulation of key enzyme genes in the plant hormone signaling pathway of T. mongolicum embryonic cells in response to stress. Conclusion Through transcriptome sequencing, it has been preliminarily demonstrated that EMS induces differential gene expression in T. mongolicum embryogenic cells and regulates the related enzyme genes of plant hormone signaling pathways in T. mongolicum embryogenic cells, in order to provide reference and basis for cultivating high-quality T. mongolicum from the molecular gene perspective.
[中圖分類(lèi)號(hào)]
R286.12
[基金項(xiàng)目]
河北省農(nóng)林科學(xué)院基本科研業(yè)務(wù)費(fèi)項(xiàng)目(2023010101);河北省重點(diǎn)研發(fā)項(xiàng)目(21326312D-8);河北省農(nóng)林科學(xué)院科技創(chuàng)新專(zhuān)項(xiàng)課題資助(2022KJCXZX-BHS-4)