| 摘要: |
| 【目的】挖掘芜菁叶绿素和叶黄素代谢关键基因，为解析芜菁果皮颜色性状的形成机制奠定基础。【方法】以白芜菁和绿芜菁肉质根果皮为材料，测定其叶绿素含量，提取芜菁肉质根果皮总RNA，构建cDNA文库，进行转录组Illumina测序，筛选差异表达基因（DEGs），进行GO和KEGG分析。提取芜菁基因组DNA，构建DNA文库，利用Hiseq X10 PE150测序平台进行测序，分析DEGs的序列变异。随机选取6个DEGs，采用RT qPCR法对其表达水平进行验证。【结果】与白芜菁相比，绿芜菁叶绿素a和叶绿素b含量较高。转录组测序共得到2.40×108条高质量片段(reads)；与白芜菁相比，绿芜菁有2 475个上调DEGs，有4 496个下调DEGs。DEGs显著富集到4 264个生物过程，712个细胞组成，1 987个分子功能的GO条目数；KEGG通路富集结果显示，差异基因显著富集到28个代谢通路中，包含代谢途径、次生代谢物生物合成、光合作用、光合作用-天线蛋白和植物激素信号转导等。与白芜菁相比，绿芜菁叶绿素代谢途径共有16个DEGs，其中尿卟啉原Ⅲ合酶（hemD）、镁螯合酶亚基H（CHLH）、联乙烯还原酶（DVR）、原叶绿素酸脂还原酶（POR）、叶绿素酶（CLH）和叶绿素a合成酶（clhG）相关基因表达上调，而共卟啉原Ⅲ氧化酶(CPX)、镁脱螯酶(SGR)、嗜铬酶(PPD)相关基因表达下调；叶黄素合成途径共鉴定到4个DEGs，其中LCYE(BraA01g027340E、BraA10g046272E)、CYP97C1(BraA07g035488E)表达量上调，而BETA-OHASE(BraA03g012-433E)表达量下调。2种芜菁在叶绿素代谢通路中，有5个DEGs(NOL、chlH、POR、chlG和CPX)有序列差异；在叶黄素合成途径中，有3个DEGs（LCYE、BETA-OHASE和CYP97C1）存在序列差异。6个 DEGs（SGR（BraA01g-025078E）、CPX（BraA09g007096E）、POR（BraA01g025517E）、chlG （BraA09g004947E）、chlH（BraA03g011972E）和POR（BraA10g044962E））表达水平的RT-qPCR结果与转录组测序结果相同。【结论】白芜菁和绿芜菁之间有16个叶绿素代谢途径相关DEGs，其中5个基因有序列差异；有3个叶黄素合成途径相关DEGs。 |
| 关键词: 芜菁 肉质根 转录组测序 叶绿素代谢 叶黄素代谢 |
| DOI：10.13207/j.jnwafu.2026.02.016 |
| 分类号: |
| 基金项目:西藏自治区重点研发项目“高海拔庭院蔬菜立体无土栽培设施研发与应用示范”（XZ202401ZY0088）；中央支持地方建设项目（533323003）；温州市基础性公益科研项目（N2023014） |
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| Investigation of genes related to chlorophyll and lutein metabolism in the fleshy root of turnip |
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WANG Yancai1,2, WANG Jiangyan1,2, XU Huanwen2, WANG Zhonghong1, FENG Xibo1
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1.College of Plant Science,Xizang Agricultural and Animal Husbandry University,Linzhi,Xizang 860000,China;2.Southern Zhejiang Key Laboratory of Crop Breeding,Wenzhou Vocational College of Science and Technology(Wenzhou Academy of Agricultural Sciences),Wenzhou,Zhejiang 325006,China
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| Abstract: |
| 【Objective】This research aimed to investigate the key genes of chlorophyll and lutein metabolism pathway,to lay a foundation for the analysis of the formation mechanism of turnip pericarp colour traits.【Method】With white turnip and green turnip fleshy root pericarps as materials,the chlorophyll content was determined,total RNA was extracted,and a cDNA library was constructed.Illumina sequencing of the transcriptome was carried out to screen differentially expressed genes (DEGs),and GO and KEGG analyses were performed.Turnip genomic DNA was extracted,and a DNA library was constructed.The Hiseq X10 PE150 sequencing platform was used for DNA sequencing to analyze the sequence variation of DEGs.Six DEGs were randomly selected and their expression levels were verified by RT-qPCR.【Result】Green turnips had higher chlorophyll a and chlorophyll b content compared to white turnips.A total of 2.40×108 high-quality reads were obtained by transcriptome sequencing.Compared with white turnip,green turnip had 2 475 up-regulated DEGs and 4 496 down-regulated DEGs.DEGs were significantly enriched to GO terms of 4 264 biological processes,712 cellular components,and 1 987 molecular functions.KEGG pathway enrichment results showed that the DEGs were significantly enriched to 28 metabolic pathways containing metabolic pathways,secondary metabolite biosynthesis,photosynthesis,photosynthesis-antenna proteins and phytohormone signal transduction.Compared with white turnip,16 DEGs in the chlorophyll metabolic pathway were screened in green turnip,including up-regulated expression genes (uroporphyrinogen Ⅲ synthase(hemD),magnesium chelatase subunit H(CHLH),diphenylethylene reductase(DVR),proto-chlorophyllinic acid lipids reductase(POR),chlorophyllotoxin lyase(CLH),and chlorophyll a synthase(clhG)) and down-regulated expression genes(coproporphyrinogen Ⅲ oxidase (CPX),magnesium dechelatase,magnesium deglutaminase (SGR),and chromophore enzyme(PPD)-related gene expression).A total of four DEGs were identified in the lutein synthesis pathway,of which LCYE (BraA01g027340E,BraA10g-046272E),and CYP97C1(BraA07g035488E) expression were up-regulated,and BETA-OHASE (BraA-03g012433E) expression was down-regulated.The two turnip species showed sequence differences in five DEGs (NOL,chlH,POR,chlG,and CPX) in the chlorophyll metabolic pathway,and three DEGs (LCYE,BETA-OHASE,and CYP97C1) in the lutein synthesis pathway.The RT-qPCR results of the expression level of six DEGs (SGR (BraA01g025078E),CPX(BraA09g007096E),POR (BraA01g0255-17E),chlG (BraA09g004947E),chlH (BraA03g011972E),and POR (BraA10g044962E)) were consistent with the transcriptome sequencing results.【Conclusion】There are 16 DEGs related to chlorophyll metabolism pathway between white turnip and green turnip,among which 5 genes show genome sequence differences.There are 3 DEGs related to lutein synthesis pathway. |
| Key words: turnips fleshy roots transcriptome sequencing chlorophyll metabolism lutein metabolism |
