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园艺学报 ›› 2023, Vol. 50 ›› Issue (10): 2117-2127.doi: 10.16420/j.issn.0513-353x.2022-0766

• 遗传育种·种质资源·分子生物学 • 上一篇    下一篇

卷丹侧生器官边界域基因LlLBD18的克隆和功能分析

殷小雨1,2, 何国仁3, 毕蒙蒙2, 唐玉超2, 郝春莲1,2, 渠雨潇2, 郝泽慧4, 徐雷锋2, 胡凤荣1,*(), 杨盼盼2,*(), 明军2,*()   

  1. 1 南京林业大学风景园林学院,南京 210037
    2 中国农业科学院蔬菜花卉研究所,蔬菜生物育种全国重点实验室,北京 100081
    3 上海师范大学生命科学院,上海 200233
    4 北京农学院园林学院,北京 102206
  • 收稿日期:2023-05-19 修回日期:2023-10-13 出版日期:2023-10-25 发布日期:2023-10-30
  • 通讯作者: *(E-mail:mingjun@caas.cn,yangpanpan@caas.cn,hufengrong2003@sina.com)
  • 基金资助:
    国家自然科学基金项目(31902043); 国家自然科学基金项目(32172612); 国家重点研发计划项目(2019YFD1001002); 中央级公益性科研院所基本科研业务费专项(IVF-BRF2022015)

Cloning and Functional Analysis of LlLBD18 in Lilium lancifolium

YIN Xiaoyu1,2, HE Guoren3, BI Mengmeng2, TANG Yuchao2, HAO Chunlian1,2, QU Yuxiao2, HAO Zehui4, XU Leifeng2, HU Fengrong1,*(), YANG Panpan2,*(), MING Jun2,*()   

  1. 1 College of Landscape Architecture,Nanjing Forestry University,Nanjing 210037,China
    2 State Key Laboratory of Vegetable Biobreeding,Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China
    3 College of Life Sciences,Shanghai Normal University,Shanghai 200233,China
    4 College of Landscape Architecture,Beijing University of Agriculture,Beijing 102206,China
  • Received:2023-05-19 Revised:2023-10-13 Published:2023-10-25 Online:2023-10-30
  • Contact: *(E-mail:mingjun@caas.cn,yangpanpan@caas.cn,hufengrong2003@sina.com)

摘要:

以卷丹(Lilium lancifolium)为试材,克隆了1个LBD转录因子基因,命名为LlLBD18(GenBank序列号ON455210)。其开放阅读框为684 bp,编码227个氨基酸,含有1个典型的LOB结构域。进化树分析发现LlLBD18与姜(Zingiber officinale)ZoLBD18的亲缘关系最近。亚细胞定位结果显示LlLBD18定位于细胞质和细胞核。荧光定量PCR分析表明,LlLBD18在卷丹初生珠芽中表达量最高,其次是根中,在叶片中最低。在珠芽形成过程中,LlLBD18的整体表达情况为先上升后下降再上升,且在珠芽原基启动前期(离体诱导4 d)最高。功能研究发现,在卷丹中过表达LlLBD18可以促进珠芽形成,而沉默LlLBD18后的珠芽形成受到显著抑制,表明LlLBD18在卷丹珠芽形成中起正向调节作用。

关键词: 卷丹, 侧生器官, 珠芽形成, 基因克隆, 功能分析

Abstract:

In this study,a LBD transcription factor gene,named LlLBD18(GenBank accession number ON455210)was cloned from Lilium lancifolium. Its open reading frame was 684 bp,which encoded 227 amino acids,and contained a typical LOB domain. The phylogenetic tree analysis showed that LlLBD18 had the highest homology with Zingiber officinale ZoLBD18. Subcellular localization results showed that LlLBD18 was localized in the cytoplasm and nucleus. Quantitative real-time PCR analysis showed that the expression of LlLBD18 was the highest in primary bulbils,followed by roots,and the lowest in leaves. In the process of bulbil formation,the overall expression of LlLBD18 was first increased,then decreased,and then increased,and the highest expression was at the early stage of bulbil primordium initiation(the 4th day). Functional studies found that overexpression of LlLBD18 in L. lancifolium can promote bulbil formation,while the formation of bulbils after VIGS silencing of LlLBD18 was significantly inhibited. This research suggested that LlLBD18 plays a positive regulatory role in bulbil formation of L. lancifolium.

Key words: Lilium lancifolium, lateral organ, bulbil formation, gene cloning, function analysis