基于转录组测序的红掌营养生长期相关基因筛选

Screening of genes related to vegetative growth stage of Anthurium andraeanum based on transcriptome sequencing

  • 摘要: 【目的】 通过转录组测序挖掘调控红掌营养生长期长短的相关基因,为了解红掌从营养生长向生殖生长转变过程中的基因调控机制及短营养生长期的红掌分子育种提供理论依据。【方法】 以进入生殖生长阶段的广花小天娇红掌和仍处于营养生长阶段的2016-035-5红掌为研究对象,采集2个营养生长期长短不同的红掌种质的根、茎、叶、芽进行转录组测序分析,筛选差异表达基因(DEGs),对DEGs进行GO功能注释和KEGG信号通路富集分析,筛选出参与调控红掌营养生长期长短的关键基因和信号通路,并通过实时荧光定量PCR验证转录组数据的可靠性。【结果】 转录组测序共获得203.18 Gb的有效数据(Clean data),从2个红掌种质中筛选出6151个DEGs,根、茎、叶、芽4个比较组共有550个DEGs。GO功能注释结果显示,DEGs被注释到的GO功能条目主要为生物过程中的代谢过程、细胞过程、单生物过程、应激响应、生物调控、生物过程调控和定位,细胞组分中的细胞部分和细胞,以及分子功能中的结合和催化活性。KEGG信号通路富集分析结果显示,DEGs富集最多的KEGG通路主要为代谢途径、次生代谢物的生物合成、植物—病原体互作、苯丙素生物合成、碳代谢和植物激素信号转导等。共筛选出42个调控红掌营养生长期长短的DEGs,其中光周期途径24个、赤霉素途径3个、温敏途径6个、年龄途径3个、自主途径2个及开花整合因子途径4个。转录组测序与实时荧光定量PCR验证结果的相关系数(R2)为0.7293,表明转录组测序数据可靠。【结论】 红掌营养生长期长短受光周期、赤霉素、温敏、年龄、自主和开花整合因子等途径调控,红掌PRR5PHLCOL5BBX24VIL2PIF4SPL10FTFD是造成红掌营养生长期及开花时间发生改变的关键基因。

     

    Abstract: 【Objective】 This study aimed to explore the genes regulating duration of vegetative growth stage of Anthuri-um andraeanum through transcriptome sequencing, so as to provide theoretical references for elucidating gene regulation mechanism during the transition from vegetative growth to reproductive growth and molecular breeding of vegetative growth stage of Anthurium andraeanum. 【Method】 Taking Anthurium andraeanum cultivar Guanghua Xiaotianjiao ente-ring the reproductive growth and Anthurium andraeanum cultivar 2016-035-5 in the vegetative growth stage as research objects, transcriptome sequencing analysis was performed on roots, stems, leaves, and buds of the two germplasms with vegetative growth stage of different durations to screen for differentially expressed genes (DEGs). The GO functional annotation and KEGG signaling pathway enrichment analysis of DEGs were performed to identify key genes and signaling pathways involved in regulating the duration of vegetative growth stage of Anthurium andraeanum. The reliability of transcriptome data was verified by real-time fluorescence quantitative PCR. 【Result】 A total of 203.18 Gb of clean data was obtained through transcriptome sequencing, and 6151 DEGs were screened from the two Anthurium andraeanum germplams. A total of 550 DEGs were identified in the four comparison groups of roots, stems, leaves, and buds. GO functional analysis showed that DEGs were mainly annotated to: biological process terms such as metabolic process, cellular process, single-organism process, response to stimulus, biological regulation, regulation of biological process process, and localization; cellular components terms including cell part and cell; molecular function terms such as binding and catalytic activity. KEGG signaling pathway enrichment analysis showed that the KEGG pathways of DEGs mainly annotated to metabolic pathway, biosynthesis of secondary metabolites, plant-pathogen interaction, phenylpropanoid biosynthesis, carbon metabolism, plant hormone signal transduction, etc. A total of 42 genes regulating duration of vegetative growth stage of Anthurium andraeanum were identified: 24 in the photoperiod pathway, 3 in the gibberellin pathway, 6 in the thermosensitive pathway, 3 in the age pathway, 2 in the autonomous pathway, and 4 in the flowering integration factor pathway. The correlation coefficient (R²) between the transcriptome sequencing and real-time fluorescence quantitative PCR results was 0.7293, indicating that the transcriptome sequencing data were reliable. 【Conclusion】 The duration of vegetative growth stage of Anthurium andraeanum is regulated by multiple pathways, including pathways of photoperiod, gibberellin, thermosensitivity, age, autonomy, and flowering integration factor. The PRR5PHLCOL5BBX24VIL2PIF4SPL10FT, and FD genes are the key genes that change the duration of vegetative growth and flowering time.

     

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