| 摘要: |
| 【目的】筛选在不同光照下甘蓝型油菜幼苗的差异表达基因和差异代谢物，为花青素合成机理的研究奠定基础。【方法】以甘蓝型油菜自交系GLH4为研究材料，选取优质种子种植于花盆，在人工气候箱中以300 μmol/(m2· s）光照培养至2叶1心期后，进行强光700 μmol/(m2·s)和弱光300 μmol/(m2· s）处理，取0,24,48,72和96 h油菜幼苗进行表型观察；分别取不同光照处理96 h油菜的第1片真叶，利用高通量测序技术获得GLH4在不同光照下的转录组数据，随后利用生物信息学方法进行差异基因KEGG分析，筛选不同光照处理油菜幼苗差异表达基因，并用RT-qPCR技术进行验证；用超高效液相色谱和串联质谱和Analyst 1.6.3软件进行油菜幼苗代谢组数据的处理，对代谢物进行定性定量分析，筛选不同光照处理的差异代谢物。【结果】对0,24,48,72和96 h油菜幼苗的表型观察发现，强光处理油菜幼苗茎、叶颜色变紫，且随诱导时间延长，颜色逐渐变深。转录组KEGG分析共找到9 840个差异表达基因，且参与光照调控的差异基因显著富集到了20条通路中，其中代谢途径的差异表达基因最多（1 751个），其次为次生代谢途径的差异表达基因（986个）。代谢组分析共检测出248个差异代谢物，其中类黄酮差异代谢物最多（81个）。类黄酮代谢物中包括7种花青素苷，其中花翠素、花翠素 3-O-葡萄糖苷、矢车菊素和牵牛花色素3-O-葡萄糖的表达量达到极显著水平。类黄酮代谢途径的差异表达基因和差异代谢物的表达量均达到了极显著水平；表达量居前的8个差异表达基因的RT-qPCR结果与转录组分析结果一致。【结论】强光诱导油菜器官和组织颜色的变化与花青素积累及相关基因的表达变化有关。 |
| 关键词: 甘蓝型油菜 花青素合成 油菜育种 |
| DOI： |
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| 基金项目:青海省科技厅项目“彩色油菜种质资源创新及利用”(2020-ZJ-745) |
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| Analysis of transcriptome and metabolome of light-induced anthocyanin synthesis in Brassica napus L. seedling |
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TENG Shoulian,LI Pan,QI Lijuan,et al
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| Abstract: |
| 【Objective】This study screened and analyzed the differentially expressed genes and metabolites in Brassica napus L.seedlings under different light conditions to provide basis for understanding anthocyanin synthesis mechanism.【Method】High quality seeds of inbred line GLH4 of Brassica napus L. were selected and planted in flowerpots.The seeds were cultured under 300 μmol/(m2·s) light in an artificial climate chamber until the stage of 2 leaves and 1 heart.Then,two different light treatments of high light 700 μmol/(m2·s) and low light 300 μmol/(m2·s) were performed.The phenotypes of rape seedlings were observed at the ages of 0,24,48,72 and 96 h.The first true leaf of rapeseed treated with different light conditions for 96 h was collected,and the transcriptome data of GLH4 was obtained by high throughput sequencing technology.Then,the KEGG differential gene analysis was performed by bioinformatics method,and two differentially expressed genes were screened and verified by RT-qPCR technology.Ultra performance liquid chromatography (UPLC-MS),tandem mass spectrometry (MS) and software Analyst 1.6.3 were used to process the metabolome data of rapeseed seedlings.The metabolites were then qualitatively and quantitatively analyzed and used for statistical analysis to screen different metabolites under the two treatments.【Result】Phenotypic observation of rapeseed seedlings at 0,24,48,72 and 96 h showed that the color of stem and leaf turned purple under strong light treatment,and it gradually darkened with the extension of induction time.A total of 9 840 differentially expressed genes were found by KEGG analysis of transcriptome,and the differentially expressed genes involved in different light treatments were significantly enriched in 20 pathways.The most differentially expressed genes were metabolic pathways (1 751),followed by secondary metabolic pathways (986).A total of 248 differential metabolites were detected by metabolome analysis with the most for flavonoids (81).Flavonoid metabolites included 7 anthocyanins,and expression levels of anthocyanin,anthocyanin 3-O-glucoside,cornulin and petunia 3-O-glucose reached extremely significant levels.Only differentially expressed genes and differentially expressed metabolites in the flavonoid metabolic pathway reached extremely significant levels.The validation results of top 8 differentially expressed genes were consistent with the results of transcriptome analysis.【Conclusion】The changes of color in organs and tissues induced by light were related with the changes of anthocyanin accumulation and related gene expression. |
| Key words: Brassica napus L. anthocyanins synthesis Brassica napus L.cultivation |
