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园艺学报 ›› 2020, Vol. 47 ›› Issue (8): 1490-1504.doi: 10.16420/j.issn.0513-353x.2019-0963

• 研究论文 • 上一篇    下一篇

‘红元宝’紫玉兰两次花芽分化差异代谢通路及关键调控基因筛选

程少禹,宣铃娟,董 彬,顾翠花,申亚梅*,张明如,戴梦怡,王卓为,章颖佳,陆丹迎   

  1. 浙江省园林植物种质创新与利用重点实验室,浙江农林大学风景园林与建筑学院,杭州 311300
  • 出版日期:2020-08-25 发布日期:2020-08-25
  • 基金资助:
    浙江省“十三五”林木育种专项(2016C02056-1);浙江省重点研发项目(2019C02023)

Identification of Differential Metabolic Pathways and Key Regulatory Genes in the Two Flower Bud Differentiation Processes of Magnolia liliiflora

CHENG Shaoyu,Xuan Lingjuan,DONG Bin,GU Cuihua,SHEN Yamei*,ZHANG Mingru,DAI Mengyi,WANG Zhuowei,ZHANG Yingjia,and LU Danying   

  1. Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants,School of Landscape Architecture,Zhejiang A & F University,Hangzhou 311300,China
  • Online:2020-08-25 Published:2020-08-25

摘要: 为了探究调控‘红元宝’紫玉兰一年两次花芽分化的成花关键基因和代谢通路,对其两次花芽分化过程的前、中、后期花芽样本进行转录组和代谢组测序分析。转录组拼接后共得到43 257条Unigene,其中鉴定出35个差异表达的成花关键基因;代谢组共检测到569个代谢物。结合转录组和代谢组分析,两次花芽分化中期产生差异基因最多,共4 074个。第二次花芽分化产生的差异代谢物蔗糖(Sucrose)和3–氰基丙氨酸(3-Cyanoalanine)大幅上调,甲基丙二酸(Methylmalonic acid)大幅下调,它们在4条KEGG通路上与相应时期的差异基因的相关性值|PCC| > 0.80且P < 0.05。蔗糖与淀粉代谢通路中,差异代谢物海藻糖(Trehalose)与该通路中7个差异基因相关性值|PCC| > 0.80。通过CCA(Canonical correspondence analysis)分析:两次花芽分化的中期,筛选出存在差异转录调控机制的KEGG通路共3条,分别为ko01200碳代谢、ko00630乙醛酸和二羧酸代谢和ko02010ABC转运蛋白。从35个成花基因中随机挑选6个(MlCOL9、MlGA9、MlGAI、MlSPL4、MlSPY、MlSVP)进行qPCR验证,其表达模式与转录组基本一致,说明转录组数据可靠。研究表明:‘红元宝’紫玉兰二次花芽分化是受到光周期途径、春化途径、年龄途径和赤霉素途径的共同影响以及8条代谢通路的协同作用而产生,且代谢物蔗糖和海藻糖在其中发挥重要的作用。

关键词: 紫玉兰, 二次成花, 差异基因, 差异代谢物, 实时荧光定量PCR

Abstract: Magnolia liliiflora‘Hongyuanbao’blooms in spring and summer,transcriptome sequencing and metabolome sequencing were analyzed to explore the differential metabolic pathways and key regulatory genes during second flower bud differentiation processes of‘Hongyuanbao’. 43 257 unigenes and 569 metabolites were identified. Thirty-five floral development genes were involved in the twice flower bud differentiation. Combine transcriptomic with metabolomics analysis,4 074 genes were involved in the middle stage of second flower bud differentiation. Meanwhile,sucrose and 3-cyanoalanine were significantly up-regulated in the second time,and methylmalonic acid was significantly down-regulated(|log2FC| ≥ 2). The correlation value of the corresponding differential genes was |PCC| > 0.80 and P < 0.05 in four KEEG pathways. The correlation between trehalose and seven differential genes were |PCC| > 0.80 in the sucrose and starch metabolic pathway. CCA analysis showed that comparison between the middle stage of twice flower bud differential process identified three KEGG pathways:ko01200 carbon metabolism,ko00630 glyoxylic acid and dicarboxylic acid metabolism and ko02010ABC transport protein. The expression of genes related to floral development,6 genes(MlCOL9,MlGA9,MlGAI,MlSPL4,MlSPY,MlSVP)were selected from 35 different genes for qRT-PCR verification,which verified the accuracy of transcriptome sequencing results. In summary,this study showed that the secondary flower bud differentiation of‘Hongyuanbao’was produced by the combined influence of photoperiod pathway,vernalization pathway,age pathway and gibberellin pathway and 8 metabolic pathways,and the sucrose and trehalose played important role in it.

Key words: Magnolia liliiflora, secondary flowering, differential gene, differential metabolite, qRT-PCR

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