银杏褐化与非褐化愈伤组织的转录组分析
Transcriptome Analysis of Browning and Non-browning Callus of Ginkgo biloba
[目的]本研究利用环境扫描电子显微镜和高通量测序技术对银杏愈伤组织进行细胞结构观察与转录组测序,以探索愈伤组织的褐化机理。[方法]利用环境扫描电子显微镜对银杏褐化和非褐化的愈伤组织进行细胞结构观察,并对银杏褐化和非褐化的愈伤组织进行转录组测序,利用DESeq2筛选差异表达基因,借助GO、KEGG等生物信息学分析手段探索基因功能。[结果]扫描电镜显示非褐变的愈伤组织细胞排列均匀,褐变愈伤组织的细胞表现出紊乱,排列松散无序。转录组测序共发现有 275和 153个基因上调和下调表达;GO分析结果显示这些基因在细胞过程、代谢过程、定位、薄膜部分、细胞部分、细胞器、催化活性、结合和转运蛋白活性等分类中显著富集;KEGG分析结果显示这些基因在苯丙烷生物合成、角质、软木质和蜡的生物合成、类黄酮生物合成以及植物-病原菌相互作用通路中显著富集,苯丙烷代谢途径中PAL和CCR基因上调表达;转录因子分析发现相较于非褐化愈伤组织,褐化愈伤组织中MYB、AP2和NAC等转录因子表达量上调,H D-ZIP下调。[结论]褐变愈伤组织的细胞排列松散甚至破裂,不能继续生长。差异表达基因显著富集在苯丙烷代谢途径,P AL 和 CCR 基因上调表达增加了酚酸的合成与代谢,推测苯丙烷生物代谢途径部分基因表达量上调导致酚酸类化合物的积累,可能是银杏愈伤组织褐化的主要原因。本研究通过扫描电镜观察和转录组测序,为解析银杏愈伤组织褐化的分子机制奠定基础。
[Objective]In this study,environmental scanning electron microscopy and high-throughput sequencing techn ology were used to observe the cell structure of Ginkgo biloba callus and sequence the transcriptome to explore the brow ning mechanism of callus.[Method]Using environmental scanning electron microscope to observe the cell structure of the browned and non-browning callus of Ginkgo biloba,and sequence the transcriptome of the browned and non-browning ca llus of Ginkgo biloba,and use DESeq2 to screen the differentially expressed genes.Use GO,KEGG and other bioinforma tics analysis methods to explore gene functions.[Result]Scanning electron microscopy showed that the cells of the non-br owning callus were evenly arranged,and the cells of the browning callus were disordered and arranged loosely and disord erly.Transcriptome sequencing found that there were 275 and 153 genes up-regulated and down-regulated;GO analysis res ults showed that these genes are classified in cellular processes,metabolic processes,localization,membrane parts,cell part s,organelles,catalytic activities,binding and transport protein activities,etc.Significant enrichment;KEGG analysis results show that these genes are significantly enriched in phenylpropane biosynthesis,keratin,soft wood and wax biosynthesis,fl avonoid biosynthesis,and plant-pathogen interaction pathways.PAL and CCR in the phenylpropane metabolism pathway Ge ne up-regulated expression;transcription factor analysis found that compared with non-browning callus,the expression of MYB,AP2 and NAC and other transcription factors in browned callus was up-regulated,and HD-ZIP was down-regulated.[Conclusion]The cell arrangement of the browned callus is loose or even broken,and cannot continue to grow.Differen tially expressed genes are significantly enriched in the phenylpropane metabolic pathway.The up-regulation of PAL and C CR genes increases the synthesis and metabolism of phenolic acid.It is speculated that the up-regulation of some genes in the phenylpropane biological metabolic pathway leads to the accumulation of phenolic acids,which may be due to the ac cumulation of phenolic acids.The main cause of browning of wounded tissue.This study lays the foundation for analyzin g the molecular mechanism of Ginkgo callus browning through scanning electron microscopy observation and transcriptome sequencing.
徐奇;杨晓明;汪贵斌;曹福亮;
Ginkgo biloba Callus Browning and non-browning Transcriptome Differentially expressed gene
2081-2090 / 10
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