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园艺学报 ›› 2021, Vol. 48 ›› Issue (10): 1859-1872.doi: 10.16420/j.issn.0513-353x.2021-0476

• 研究论文 • 上一篇    下一篇

红掌R2R3-MYB转录因子基因AaMYB6调控花青素苷合成

李崇晖1,3, 杨光穗1,3, 张志群1,3, 尹俊梅2,3,*()   

  1. 1中国热带农业科学院热带作物品种资源研究所 农业部华南作物基因资源与种质创制重点实验室,海口 571101
    2中国热带农业科学院海口实验站,海口 571101
    3海南省热带观赏植物种质创新利用工程技术研究中心,海南儋州 571737
  • 收稿日期:2021-07-19 修回日期:2021-09-02 出版日期:2021-10-25 发布日期:2021-11-01
  • 通讯作者: 尹俊梅 E-mail:yinjunmei2004@163.com
  • 基金资助:
    海南省自然科学基金创新研究团队项目(2018CXTD344);中国热带农业科学院基本科研业务费专项资金项目(1630032017024)

A Novel R2R3-MYB Transcription Factor Gene AaMYB6 Involved in Anthocyanin Biosynthesis in Anthurium andraeanum

LI Chonghui1,3, YANG Guangsui1,3, ZHANG Zhiqun1,3, YIN Junmei2,3,*()   

  1. 1Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences;Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China,Ministry of Agriculture,Haikou 571101,China
    2Haikou Experimental Station,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China
    3The Engineering Technology Research Center of Tropical Ornamental Plant Germplasm Innovation and Utilization,Danzhou,Hainan 571737,China
  • Received:2021-07-19 Revised:2021-09-02 Online:2021-10-25 Published:2021-11-01
  • Contact: YIN Junmei E-mail:yinjunmei2004@163.com

摘要:

为了挖掘调控红掌肉穗花序花青素苷合成的转录因子基因,基于前期的转录组数据,以‘粉冠军’红掌(Anthurium andraeanum‘Pink Champion’)肉穗花序为材料,克隆了1个R2R3-MYB家族基因AaMYB6(GenBank序列号MZ171064)。通过氨基酸序列比对和系统进化树分析、亚细胞定位、在烟草中的过表达分析、qRT-PCR、酵母双杂交(yeast two-hybrid assay,Y2H)和双分子荧光互补(bimolecular fluorescence complementation,BiFC)试验,研究了AaMYB6的功能。结果表明AaMYB6与已报道的AaMYB1具有较高的同源性,属于以玉米ZmC1为代表的,调控单子叶植物花青素苷合成的R2R3-MYB家族C1亚组。AaMYB6定位于细胞核。在烟草中过表达AaMYB6,NtF3H表达量下调,但却激活了T1代转基因植株花丝中花青素苷生物合成途径关键酶基因NtF3′HNtDFRNtANSNtUFGT的表达,产生了花丝中积累花青素苷的表型。AaMYB6主要在红掌‘紫公主’的肉穗花序中表达,且在不同品种肉穗花序中其表达量与花青素苷含量显著正相关。Y2H和BiFC试验表明AaMYB6与前期报道的参与花青素苷和原花青素合成调控的bHLH转录因子AabHLH1有相互作用,进一步验证了AaMYB6调控花青素苷合成的功能。

关键词: 红掌, MYB转录因子, 花青素苷, 异源过表达, 转录调控, 基因功能

Abstract:

A new R2R3-MYB gene AaMYB6 with GenBank accession number MZ171064 was isolated from the spadix of Anthurium andraeanum‘Pink Champion’based on the information from previous transcriptome data for identification of transcription factor genes associated with anthocyanin biosynthesis. The function of AaMYB6 was analyzed through the assays such as amino acid sequence alignment,phylogenetic tree analysis,subcellular localization,overexpression in tobacco,qRT-PCR,yeast two-hybrid assay(Y2H),and bimolecular fluorescence complementation(BiFC). The results suggested that AaMYB6 showed homology with previously identified AaMYB1 from anthurium and belong to C1 subgroup R2R3-MYBs,represented by Zea may C1,that regulate anthocyanin biosynthesis in monocots. AaMYB6 was proved to be nucleus-localized protein. Overexpression of AaMYB6 in tobacco markedly activated the expression of the enzyme genes NtF3′H,NtDFR,NtANS,NtUFGT,specifically involved in anthocyanin biosynthesis in filaments and promoted the accumulation of anthocyanin pigments. AaMYB6 was mainly expressed in the spadix of‘Rapido’. And the expression of AaMYB6 was closely linked to anthocyanin accumulation in the spadix of various cultivars. The interaction between AaMYB6 and the AabHLH1 protein,which was identified previously to regulate the biosynthesis of anthocyanin and proanthocyanidin in anthurium,was confirmed by Y2H and BiFC assays. The results indicated that AaMYB6 play a role in the regulation of anthocyanin biosynthesis in anthurium.

Key words: Anthurium andraeanum, MYB transcription factor, anthocyanin, heterologous overexpression, transcriptional regulation, gene function

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